海生素对肝癌的抑制作用及机制研究
|
摘要
肝细胞癌(Heptocellular carcinoma,HCC)是发病率最高的原发性肝脏恶性肿瘤。尽管目前对肝癌可以采取手术切除、动脉插管化疗、放疗及局部治疗等方法,但效果欠佳,人们一直在寻找一种有效治疗肝癌的措施。恶性肿瘤的形成与肿瘤细胞增殖、凋亡调控信号失衡有关,诱导肝癌细胞凋亡是治疗肝癌的重要策略。此外,肝癌血供非常丰富,血管生成对肿瘤的生长、侵袭和转移都有决定性的作用,而且在发病早期就有血管侵犯和肝内转移的倾向,这是肝癌预后差的重要原因。诱导肝癌细胞凋亡和抑制肝癌新生血管形成将是治疗肝癌的重要途径。
海生素是自海洋贝类毛蚶(Tegillarca granosa Linnaeus)中提取出的抗肿瘤活性物质,在课题组以往的研究中得知海生素对白血病细胞K562、肺癌细胞A549和肾癌细胞OS-RC-2有抑制作用,并能抑制小鼠艾氏腹水瘤。而海生素诱导肝癌细胞的凋亡,以及抑制肝癌新生血管形成则是本次研究的重点。
首先,观察海生素对体外肝癌细胞生长的抑制作用,通过对细胞凋亡相关分子、细胞信号转导相关分子的检测探讨其可能的效应机制。以MTT法检测48h不同浓度海生素对人肝癌细胞BEL-7402增殖的OD值,根据OD值计算细胞生长抑制率,实验结果表明海生素能剂量依赖性抑制肝癌细胞BEL-7402的增殖。电镜结果提示海生素能明显诱导BEL-7402细胞凋亡,出现染色质浓缩、碎裂,线粒体空泡化及凋亡小体等凋亡特征性改变。100μg/ml海生素作用48h后,流式细胞仪PI染色凋亡率为29.63±1.09%,能明显诱导细胞凋亡；Hoechst 33258荧光染色法呈现出凋亡核固缩形态,可见典型的凋亡小体。在证明凋亡发生的基础上,进一步进行海生素诱导凋亡的机制研究。RT-PCR和Western Blot检测海生素作用后BEL-7402细胞Fas mRNA和蛋白的表达,随着海生素浓度增加FasmRNA以及蛋白表达量增多。提示海生素诱导BEL-7402凋亡可能和Fas表面受体通路有关。进一步用western blot方法检测其下游信号分子的表达和活化情况,结果表明海生素能引起caspase-8和caspase-3的活化,以及细胞色素C的释放；而用caspase-8抑制剂z-IETD-fmk事先孵育后,caspase-8和caspase-3的活化片段明显减少,表明海生素可能通过Fas通路及线粒体信号通路诱导肝癌细胞BEL-7402的凋亡。
其次,由于体外环境的局限性,应用荷瘤裸鼠动物模型进一步研究海生素体内抑瘤效应及其可能的作用机制。用皮下接种BEL-7402细胞的方法建立了裸鼠荷瘤模型,62.5mg/kg、125mg/kg、250mg/kg海生素肠溶胶囊组的抑瘤率分别为50%、58%和60%,相对肿瘤增殖率分别为45%、42%和32%,表明海生素肠溶胶囊对荷人肝癌裸鼠移植瘤BEL-7402的生长有明显的抑制作用。HE结果显示,海生素组的肿瘤组织出现细胞凋亡和坏死现象,电镜结果表明海生素引起移植瘤组织中的肝癌细胞发生凋亡的特征性改变,从而从形态学证实海生素作用后肿瘤细胞发生了凋亡。原位杂交技术检测到移植瘤组织中Fas mRNA的表达在海生素用药后明显增加,免疫组织化学法检测到移植瘤组织中caspase-8、caspase-3蛋白表达增加。进一步证明,海生素通过Fas信号通路诱导BEL-7402细胞凋亡。
最后,观察海生素对肝癌新生血管形成的抑制作用。以人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)为研究对象,用MTT实验结果表明海生素抑制HUVEC的增殖活性,并且用Transwell小室法和管腔形成实验观察到海生素能明显抑制HUVEC的迁移和管腔形成,表明海生素能够直接抑制内皮细胞。体内试验选用鸡胚尿囊膜实验,观察到海生素对鸡胚尿囊膜新生血管也有明显的抑制作用。微血管密度(microvessel density,MVD)是反映肿瘤血管形成的比较直观的指标,用CD34分子免疫组织化学法观察到海生素用药组裸鼠移植瘤组织MVD明显减少。与血管形成有关的血管内皮细胞生长因子(VEGF)以及基质金属蛋白酶-2(MMP-2)的免疫组化蛋白表达在海生素用药组都明显降低。提示,海生素不仅能直接抑制血管内皮细胞,还能通过影响VEGF和MMP-2的表达而影响肝癌新生血管的形成。
总之,海生素对肝癌的抑制作用是多方面的,不仅通过Fas信号通路诱导肝癌细胞凋亡发挥其抗肿瘤活性,并且可以靶向血管具有抑制肝癌新生血管形成的作用,从而多靶点、多途径抑制肝细胞癌。
Hepatocellular carcinoma (HCC) is the most common primary liver malignancy. The current treatment options for liver cancer, including liver resection surgery, intra-arterial chemotherapy and radiation therapy, remain disappointing. Scientists are working hard, however, to address this problem.As carcinogenesis is closely related to the control of tumor cell proliferation, and apoptosis, the induction of apoptotic cell death may be the important therapeutic strategy for live cancer treatment. Meanwhile, angiogenesis plays a critical role in tumor growth, invasion and metastasis. Live cancer is rich in blood supply, its high mortality rate is mainly a result of vascular invasion and intra-hepatic metastases occurring during the early stage of cancer formation.
Haishengsu, which is extracted from Tegillarca granosa Linnaeus, is an novel antineoplastic agent. Our previous studies showed that Haishengsu can inhibit the growth of human erythroleukemia K562 cell, lung adenocarcinoma epithelial A549 cell line, renal cell carcinoma OS-RC-2 cell, and suppress Ehrlich ascites carcinoma in mice. The aim of the present study is to investigate the apoptotic effect and anti-angiogenesis actions of Haishengsu on hepatocellular carcinoma cells.
First, we investigated the inhibitory effect of Haishengsu on the growth of hepatocellular carcinoma cells in vitro and its effect on the apoptosis pathway. MTT metabolism assay showed that Haishengsu treatment for 72h significantly inhibited the human hepatoma BEL-7402 cell proliferation in a dose dependent manner. Electron microscopy analysis was performed to bserve apoptosis of BEL-7402 cells induced by HSS.In these cells, the typical condensed chromatin pulled away from the nuclear envelope and fragmented into several micronuclei.In addition, the nuclear volume decreased markedly and a considerable increase in vacuolisation was observed.FACS analysis showed that treatment with 100μg/ml of Haishengsu for 48h could induce 29.63±1.09% apoptosis. Hoechst 33258 staining also showed that Haishengsu caused condensed nuclei and apoptotic bodies.These data indicated that Haishengsu could induce BEL-7402 cells apoptosis. To reveal its mechanism of apoptosis induction, RT-PCR was performed to test Fas mRNA levels in BEL-7402 cells. Fas mRNA was upregulated by Haishengsu, which indicated that Fas receptor pathway may involve in Haishengsu-induced apoptosis. Western blot assay showed that haishengsu activated caspase-8 and caspase-3.Pretreatment with z-IETD-fmk markedly decreased the activation of caspase-8 and caspase-3.These data suggested that haishengsu may induce BEL-7402 apoptosis via Fas signaling pathway.
Next, based on the antiapoptotic effects of Haishengsu, we measured the antitumor activity of haishengsu in null mice inoculated with BEL-7402 cells. Daily feed with 62.5 125 and 250mgkg-1 Haishengsu for 25 days yielded tumor inhibition of about 50%,58% and 60% respectively, relative rate of tumor proliferation was 45%、42% and 32% respectively. This indicated that HSS significantly inhibited tumor growth in vivo.HE staining showed that apoptotic bodies and necrosis appeared in tumor in Haishengsu treated groups.Electron microscopy analysis showed that haishengsu caused the typical apoptotic morphology in tumor. Haishengsu increased Fas mRNA level in tumor, as assessed by in-situ hybridization. Immunohistochemistry assay showed that haishengsu increased caspase-8 and caspase-3 in mice turmor. These results confirmed that haishengsu induced BEL-7402 apoptosis via Fas singnaling pathway.
Finally, the antiangiogenic properties of haishengsu were investigated with several in vitro and in vivo models. Possible mechanisms underlying its antiangiogenic activity were also assessed. Haishengsu dose-dependently inhibited proliferation of human umbilical vein endothelial cells (HUVEC).In tube formation and cell migration assays using HUVEC cells, haishengsu significantly inhibited formation of intact tube networks and reduced the number of migratory cells. In the chick chorioallantoic membrane assay, haishengsu reduced new vessel formation compared with the vehicle control. Microvessel density (MVD) in sections of the tumor was reduced after Haishengsu treatment. The levels of VEGF, and MMP-2 were also significantly decreased.These data suggested that Haishengsu could directly inhibit vescular endothelial cells and exert anti-angiogenic effect via downregulation of VEGF and MMP-2.
Collectively, Haishengsu could inhibit hepatocellular carcinoma and its effect associated at least in part with Fas singnaling pathway and the antiangiogenesis.
海生素是自海洋贝类毛蚶(Tegillarca granosa Linnaeus)中提取出的抗肿瘤活性物质,在课题组以往的研究中得知海生素对白血病细胞K562、肺癌细胞A549和肾癌细胞OS-RC-2有抑制作用,并能抑制小鼠艾氏腹水瘤。而海生素诱导肝癌细胞的凋亡,以及抑制肝癌新生血管形成则是本次研究的重点。
首先,观察海生素对体外肝癌细胞生长的抑制作用,通过对细胞凋亡相关分子、细胞信号转导相关分子的检测探讨其可能的效应机制。以MTT法检测48h不同浓度海生素对人肝癌细胞BEL-7402增殖的OD值,根据OD值计算细胞生长抑制率,实验结果表明海生素能剂量依赖性抑制肝癌细胞BEL-7402的增殖。电镜结果提示海生素能明显诱导BEL-7402细胞凋亡,出现染色质浓缩、碎裂,线粒体空泡化及凋亡小体等凋亡特征性改变。100μg/ml海生素作用48h后,流式细胞仪PI染色凋亡率为29.63±1.09%,能明显诱导细胞凋亡；Hoechst 33258荧光染色法呈现出凋亡核固缩形态,可见典型的凋亡小体。在证明凋亡发生的基础上,进一步进行海生素诱导凋亡的机制研究。RT-PCR和Western Blot检测海生素作用后BEL-7402细胞Fas mRNA和蛋白的表达,随着海生素浓度增加FasmRNA以及蛋白表达量增多。提示海生素诱导BEL-7402凋亡可能和Fas表面受体通路有关。进一步用western blot方法检测其下游信号分子的表达和活化情况,结果表明海生素能引起caspase-8和caspase-3的活化,以及细胞色素C的释放；而用caspase-8抑制剂z-IETD-fmk事先孵育后,caspase-8和caspase-3的活化片段明显减少,表明海生素可能通过Fas通路及线粒体信号通路诱导肝癌细胞BEL-7402的凋亡。
其次,由于体外环境的局限性,应用荷瘤裸鼠动物模型进一步研究海生素体内抑瘤效应及其可能的作用机制。用皮下接种BEL-7402细胞的方法建立了裸鼠荷瘤模型,62.5mg/kg、125mg/kg、250mg/kg海生素肠溶胶囊组的抑瘤率分别为50%、58%和60%,相对肿瘤增殖率分别为45%、42%和32%,表明海生素肠溶胶囊对荷人肝癌裸鼠移植瘤BEL-7402的生长有明显的抑制作用。HE结果显示,海生素组的肿瘤组织出现细胞凋亡和坏死现象,电镜结果表明海生素引起移植瘤组织中的肝癌细胞发生凋亡的特征性改变,从而从形态学证实海生素作用后肿瘤细胞发生了凋亡。原位杂交技术检测到移植瘤组织中Fas mRNA的表达在海生素用药后明显增加,免疫组织化学法检测到移植瘤组织中caspase-8、caspase-3蛋白表达增加。进一步证明,海生素通过Fas信号通路诱导BEL-7402细胞凋亡。
最后,观察海生素对肝癌新生血管形成的抑制作用。以人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)为研究对象,用MTT实验结果表明海生素抑制HUVEC的增殖活性,并且用Transwell小室法和管腔形成实验观察到海生素能明显抑制HUVEC的迁移和管腔形成,表明海生素能够直接抑制内皮细胞。体内试验选用鸡胚尿囊膜实验,观察到海生素对鸡胚尿囊膜新生血管也有明显的抑制作用。微血管密度(microvessel density,MVD)是反映肿瘤血管形成的比较直观的指标,用CD34分子免疫组织化学法观察到海生素用药组裸鼠移植瘤组织MVD明显减少。与血管形成有关的血管内皮细胞生长因子(VEGF)以及基质金属蛋白酶-2(MMP-2)的免疫组化蛋白表达在海生素用药组都明显降低。提示,海生素不仅能直接抑制血管内皮细胞,还能通过影响VEGF和MMP-2的表达而影响肝癌新生血管的形成。
总之,海生素对肝癌的抑制作用是多方面的,不仅通过Fas信号通路诱导肝癌细胞凋亡发挥其抗肿瘤活性,并且可以靶向血管具有抑制肝癌新生血管形成的作用,从而多靶点、多途径抑制肝细胞癌。
Hepatocellular carcinoma (HCC) is the most common primary liver malignancy. The current treatment options for liver cancer, including liver resection surgery, intra-arterial chemotherapy and radiation therapy, remain disappointing. Scientists are working hard, however, to address this problem.As carcinogenesis is closely related to the control of tumor cell proliferation, and apoptosis, the induction of apoptotic cell death may be the important therapeutic strategy for live cancer treatment. Meanwhile, angiogenesis plays a critical role in tumor growth, invasion and metastasis. Live cancer is rich in blood supply, its high mortality rate is mainly a result of vascular invasion and intra-hepatic metastases occurring during the early stage of cancer formation.
Haishengsu, which is extracted from Tegillarca granosa Linnaeus, is an novel antineoplastic agent. Our previous studies showed that Haishengsu can inhibit the growth of human erythroleukemia K562 cell, lung adenocarcinoma epithelial A549 cell line, renal cell carcinoma OS-RC-2 cell, and suppress Ehrlich ascites carcinoma in mice. The aim of the present study is to investigate the apoptotic effect and anti-angiogenesis actions of Haishengsu on hepatocellular carcinoma cells.
First, we investigated the inhibitory effect of Haishengsu on the growth of hepatocellular carcinoma cells in vitro and its effect on the apoptosis pathway. MTT metabolism assay showed that Haishengsu treatment for 72h significantly inhibited the human hepatoma BEL-7402 cell proliferation in a dose dependent manner. Electron microscopy analysis was performed to bserve apoptosis of BEL-7402 cells induced by HSS.In these cells, the typical condensed chromatin pulled away from the nuclear envelope and fragmented into several micronuclei.In addition, the nuclear volume decreased markedly and a considerable increase in vacuolisation was observed.FACS analysis showed that treatment with 100μg/ml of Haishengsu for 48h could induce 29.63±1.09% apoptosis. Hoechst 33258 staining also showed that Haishengsu caused condensed nuclei and apoptotic bodies.These data indicated that Haishengsu could induce BEL-7402 cells apoptosis. To reveal its mechanism of apoptosis induction, RT-PCR was performed to test Fas mRNA levels in BEL-7402 cells. Fas mRNA was upregulated by Haishengsu, which indicated that Fas receptor pathway may involve in Haishengsu-induced apoptosis. Western blot assay showed that haishengsu activated caspase-8 and caspase-3.Pretreatment with z-IETD-fmk markedly decreased the activation of caspase-8 and caspase-3.These data suggested that haishengsu may induce BEL-7402 apoptosis via Fas signaling pathway.
Next, based on the antiapoptotic effects of Haishengsu, we measured the antitumor activity of haishengsu in null mice inoculated with BEL-7402 cells. Daily feed with 62.5 125 and 250mgkg-1 Haishengsu for 25 days yielded tumor inhibition of about 50%,58% and 60% respectively, relative rate of tumor proliferation was 45%、42% and 32% respectively. This indicated that HSS significantly inhibited tumor growth in vivo.HE staining showed that apoptotic bodies and necrosis appeared in tumor in Haishengsu treated groups.Electron microscopy analysis showed that haishengsu caused the typical apoptotic morphology in tumor. Haishengsu increased Fas mRNA level in tumor, as assessed by in-situ hybridization. Immunohistochemistry assay showed that haishengsu increased caspase-8 and caspase-3 in mice turmor. These results confirmed that haishengsu induced BEL-7402 apoptosis via Fas singnaling pathway.
Finally, the antiangiogenic properties of haishengsu were investigated with several in vitro and in vivo models. Possible mechanisms underlying its antiangiogenic activity were also assessed. Haishengsu dose-dependently inhibited proliferation of human umbilical vein endothelial cells (HUVEC).In tube formation and cell migration assays using HUVEC cells, haishengsu significantly inhibited formation of intact tube networks and reduced the number of migratory cells. In the chick chorioallantoic membrane assay, haishengsu reduced new vessel formation compared with the vehicle control. Microvessel density (MVD) in sections of the tumor was reduced after Haishengsu treatment. The levels of VEGF, and MMP-2 were also significantly decreased.These data suggested that Haishengsu could directly inhibit vescular endothelial cells and exert anti-angiogenic effect via downregulation of VEGF and MMP-2.
Collectively, Haishengsu could inhibit hepatocellular carcinoma and its effect associated at least in part with Fas singnaling pathway and the antiangiogenesis.
引文
1 World Health Organization. Mortality database. WHO Statistical Information System. Available at:http://www.who.int/whosis Accessed March 2009.
2汤钊猷.肝癌临床研究现状,2008.
3徐克.我国肝细胞癌TACE治疗的现状与进展.2008.
4 Endo K.Ueda T,Ueyama J,Ohta T, Terada T. lmmunoreactive E-cadherin,aipha-catenin, beta-catenin,and gamma-catenin proteins in hepatocellular carcinoma: Relationships with tumor grade,clinicopathologic parameters,and patients'survival. Hum.Pathol, 2003,31(5):558-565.
5 Ulmer SC. Hepatocellular carcinoma:a concise guide to its status and management. Postgrad Med,2000,107:117-124.
6 Shaw RJ,Cantley LC. Ras,PI(3)K and mTOR signalling controls tumour cell growth. Nature. 441(7092),2006.424-430.
7 Michor F,lwasa Y Nowak MA.Dynamics of cancer progression. Nat Rev Cancer, 2004,4(3):197-205.
8 Mangues R Symmans WF, Lu S,Schwartz S,Pellicer A. Activated N-ras oncogene and N-ras proto-oncogene act through the same pathway for in vivo tumorigenesis..Oncogene, 1996,13(5):1053-1063.
9 Prochownik EV. c-Myc:linking transformation and genomic instability. Curr Mol Med, 2008,8(6):446-458.
10 Hainaut P,Hollstein M. p53 and human cancer:the fa st ten thousand mutations. Adv Cancer Res,2000,77:81-137.
11 Minamoto T,Buschmann T,Habelhah H,Matusevich E,Taham H,Boerresen-Dale AL,Harris C,Sidransky D,Ronai Z. Distinct pattern of p53 phosphorylation in human tumors. Oncogene,2001,20(26):3341-3347.
12 Wang SP,Zhou HJ,Chen XP, Ren GY,Ruan XX,Zhang Y,Zhang RL,Chen J. Loss of expression of Kruppel-like factor 6 in primary hepatocellular carcinoma and hepatoma cell lines. J Exp Clin Cancer Res,2007,26(1):117-124.
13 Natoli G,Iamfi A, Costanzo A,et al.Resistance to Fas-mediated apoptosis in human hepatoma cells. Oneogene.11,1991.1157-1164.
14 Gulbins E,Jelde A,Ferlinz K, et al. Physiology of apoptosis. Am J Physiol Renal Physiol, 2000,279:605-615.
15 Carrington PE, Sandu C, Wei Y, et al. The structure of FADD and its mode of interaction with procaspase-8.Mol Cell,2006,22(5):599-610.
16 Tmmer T, Devries EG, DE Jong S. Fas receptor-mediated apoptosis:a clinical application. J Pathol,2003,15(11):983-992.
17 Donepudi M,Mac Sweeney A,Briand C,et al.Insights into the regulatory mechanism for caspase-8 activation. Mol Cell,2003,11:543-549.
18 Rodier F,Berlr R, Bossolasco M,et al. Polyomavims large T-antigen protects mouse cells
from Fas-,TNF-a and toxol-induced apoptosis. Oncogen,2000,19:6261-6270.
19 Liu A, Han X, Zhang G, Zhao X, Zheng G. Effects of cartilage polysaccharide on apoptosis of human hepatoma BEL-7402 cells and murine H22 hepatocarcinoma. Int J Food Sci Nutr, 2009,31:1-12.
20张晨,李柏,吕书勤,李勇,苏永华,凌昌全.蜂毒素对人肝癌细胞线粒体膜蛋白7A6和凋亡相关基因产物Fas及其配体FasL表达的影响.中西医结合学报,2007,5(5)：559-563.
21 Wu LQ, Zhang WJ, Niu JX, Ye LY,Yang ZH,Grau GE and Lou JN. Phenotypic and functional differences between human liver Barlcer endothelial cells and liver sinusoidal endothelial cells. J Vase Res,2008,45:78-86.
22 Nelson A, Fingleton B,Rothenberg ML,and Matrisian LM. Matrix metalloproteinases: Biologic activity and clinical implications. J.Ciin.Oneol,2000,18(5):1135-1149.
23 Livak KJ,Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods,2001,25(4):402-408.
24 S.Shen, W. Li, M. Ouyang, Z. Wu, Q. Lin, L. Xie. Identification of two marine fungi and evaluation of their antivirus and antitumor activities. Wei Sheng Wu Xue Bao, 2009,49:1240-1246.
25 U.W. Hawas, M. Shaaban, K.A. Shaaban, M. Speitling, A. Maier, G. Kelter, H.H.Fiebig, M. Meiners, E. Helmke, H. Laatsch. Mansouramycins A-D, cytotoxic isoquinolinequinones from a marine streptomycete. J Nat Prod,2009,72:2120-2124.
26 P.J. Schupp, C. Kohlert-Schupp, S. Whitefield, A. Engemann, S. Rohde, T. Hemscheidt, J.M. Pezzuto, T.P. Kondratyuk, E.J. Park, L. Marler, B. Rostama, A.D. Wright. Cancer chemopreventive and anticancer evaluation of extracts and fractions from marine macro-and microorganisms collected from Twilight Zone waters around Guam. Nat Prod Commun, 2009,4:1717-1728.
27 Li GY, Liu JZ, Yu XM, Chen SF, Zhang B, Zhang WF, et al.Effect of a seashell protein Haishengsu on cell growth and expression of apoptosis genes in leukemia K562 cells. Clin Invest Med,2008,31:E218-E221.
28 Ji-Zhu Liu, Shou-Guo Chen,Zhang Bin,Chun-Bo Wang, Guang-Yao Li,Le-Xin Wang. Antitumor effect of the seashell protein Haishengsu on Ehrlich ascites tumor:an experimental study. J Nat Med,2009,63:459-462.
29张成喜,李光耀,韩春山,叶小坤,陈洪英,周红娟,陈守国..泥蚶提取物-海生素制剂治疗中、晚期非小细胞肺癌和晚期肾癌的临床观察.中国海洋药物.24(4),2005.55-58.
30李光耀,张成喜,辛艳春,周红娟,张新,李花兰,王春波,陈守国..海生素制剂对非小细胞肺癌化疗患者生存质量的影响.中国海洋药物,2005,24(6)：50-51.
31 Li GY, Yu XM, Zhang HW, Zhang B, Wang CB, Xin YC, Yang CZ, et al. Haishengsu as an adjunct therapy to conventional chemotherapy in patients with non-small cell lung cancer:a pilot randomized and placebo-controlled clinical trial. Complement Ther Med,2009,17(1):51-55.
32 Lambert C,Landau A M,Desbarats J. Fas-beyond death:a regenerative role for Fas in the nervous system. Apoptosis,2003,8(551-562).
33 Li-Weber M,Krammer PH. Function and regulation of the CD95(APO-1/Fas) ligand in the immune system. Semin Immunol,2003,15:145-157.
34 Wajant H,Pfizenmaier K, Scheurich P. Tumor necrosis factor signaling. Cell Death Differ, 2003,10:45-63.
35 Anastassiou G,Coupland S E,Stang A,et al.Expression of Fas and Fas ligand in uveal
melanoma:biological implication and prognostic value. J Pathol,2001,194:466-472.
36 Silvestris F,Tucci M,Cafforio P,et al.Fas-L-up-regulation by highly malignant myelomant plasma cells:role in the pathogenesis of anemia and disease progression. Blood, 2001,97:1155-1164.
37 Korkolopoulou P,Goudopoulou A,Vout sinas G,et al.c-FLIP expression in bladder urothelial carcinomas:it's role in resistance to Fas-mediated apoptosis and clinicopat hologic correlations. U-rology,2004,63(1198-1204).
38 Strand S,Hofmann W J,Hug H,et al.Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing tumor cells-a mechanism of immunle evasion. N at Med,1996,2:1361-1366.
39 Nagao M,Nakajima Y,Hisanaga M,et al.The alteration of Fas receptor and ligand system in hepatocellular carcinomas:how do hepatoma cells escape from the host immune surveillance in vivo. J Hepatol,1999,30:413-421.
40 Y Cao, RW Ji, D Davidson, et al. Kringle domains of human angiostatin characterization of the anti-proliferative activity on endothelial cells. J Biol Chem,1996,271:29461-29467.
41 T Mosmann. Rapid colorimetric assay for cellular growth and survival:Application to proliferation and cytotoxicity assays. Journal of Immunological Methods,1983,65(1-2):55-63.
42 Y Cao, A Chen, SSA An, et al. Kringle 5 of Plasminogen is a Novel Inhibitor of Endothelial Cell Growth. J Biol Chem,1997,272:22924-22928.
43 T. Mitsuishi, Y. Iwabu, K. Tokunaga, T. Sata, T. Kaneko, K. Ohara, I. Ohsawa, F. Oda, Y. Yamada, S. Kawana, K. Ozaki, M. Nakatake, O. Yamada. Combined analysis of cell growth and apoptosis-regulating proteins in HPVs associated anogenital tumors. BMC Cancer,2010,10:118.
44 E.L. Kim, R. Wustenberg, A. Rubsam, C. Schmitz-Salue, G. Warnecke, E.M. Bucker, N. Pettkus, D. Speidel, V. Rohde, W. Schulz-Schaeffer, W. Deppert, A. Giese. Chloroquine activates the p53 pathway and induces apoptosis in human glioma cells. Neuro Oncol,2010,12:389-400.
45 S.Senthivinayagam, P. Mishra, S.K. Paramasivam, S.Yallapragada, M. Chatterjee, L. Wong, A. Rana, B.Rana. Caspase-mediated cleavage of beta-catenin precedes drug-induced apoptosis in resistant cancer cells. J Biol Chem,2009,284:13577-13588.
46 Kerr JF,Wyllie AH,Cullrie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br. J.Cancer,1972,26:239-257.
47 Wyllie AH,Kerr JF,Currie AR. Cell death in the significance of apoptosis. Int. Rev.Cytol,1980,68:251-306.
48 Dcbatin K M,Krammer P H. Death reception in chemotherapy and Cancer. Oncogene, 2004,23:2950-2966.
49 Ashkenazi A,Dixit V M. Death Receptors:Signaling and Modulation. Science, 1998,281:1305-1308.
50 Q. Chen, J.H. Zhang, J.T. Chen, Z.J. Shi, Y.J. Deng. MTT assay for detecting osteosarcoma cell apoptosis. Nan Fang Yi Ke Da Xue Xue Bao,2009,29:1899-1901.
51 Wesche-Soldato D E,Swan R Z,Chung C S,et al. The apoptotic pathway as a therapeutic target in sepsis. Curt Drug Targets,2007,8:493-500.
52 Everett H,McFadden G. Apoptosis:An innate immune response to virus infection. Trends Microbiol,1997,7:160-165.
53 Los M,Burek C J,Stroh C,et al.Anticancer drugs of tomorrow:Apoptotic pathways as targets for drug design. Drug Discov Today,2003,8:67-77.
54 Thompson C B. Apoptosis in the pathogenesis and treatment of disease. Science,
1995,267:1456-1462.
55 Thorbum J,Moore F,Rao A,et al.Selective Inaction of a Fas-associated Death Domain Protein (FADD)-dependent Apoptosis and Autophagy Pathway in Immon Epithelial Cells. Cells Mol Biol Cell,2005,16:1189-1199.
56 Bamhart B C,Alappat E C,Peter M E. The CD95 type Ⅰ/type Ⅱ model.Semin Immunol, 2003,15:185-193.
57 Kulms D, Zeise E, Poppelmann B, Schwarz T. DNA damage, death receptor activation and reactive oxygen species contribute to ultraviolet radiation-induced apoptosis in an essential and independent way. Oncogene,2002,196:5844-5851.
58 D.A.Martin,R.M.Siegel,L.Zheng,M.J.Lenardo. Membrane oligomerization and cleavage activates the c aspase-8(FLICE/MACHal)death signal. J.Biol.Chem,1998,274:4345-4349.
59 Bamhart B C,Alappat E C,Peter M E. The CD95 type Ⅰ/type Ⅱ model. Semin Immunol, 2003,15:185-193.
60 Boatfight K M,Salvesen G S. Mechanisms of caspase activation. Curt Opin Cell Biol, 2003,15:725-731.
61 Luo X,Budihardjo I,Zou H,Slaughter C,Wang X. Bid,a Bcl-2 interacting protein,mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell,1998,94:481-490.
62 Galvan V, Chen S, Lu D, et al. Caspase cleavage of members of the amyloid precursor family of proteins. J Neurochem.
63汪谦,主编.现代医学实验方法.北京：民卫生出版社：北京,1998.1094.
64 Canis M,Botchorishvili R,Wattiez A,et al. Tumor growth and dissemination after laparotomy and C02 pneumoperitoneum:a rat ovarian,cancer model.Obstet Gynecol, 1998,92(1):104-108.
65周际昌.实用肿瘤内科学.京：人民卫生出版社,1998.82-94.
66 Hu L,Hofmann J,Lu Y,et al.Inhibition of phosphatidylinositol 3'-kinase increases efficacy of pacl itaxel in vitro and in vivo ovarian cancer models. Cancer Res,2002,62(4):1087-1092.
67魏泓.医学实验动物学.成都：四川科学出版社,1998.302-303.
68司徒振强,吴军正.细胞培养.北京：世界图书出版社,1996.193.
69 Morinaga S, Ishiwa N, Noguchi Y, Yamamoto Y, Rino Y, Imada T, Takanashi Y, Akaike M, Sugimasa Y, Takemiya S. Growth index, assessed with Ki-67 and ssDNA labeling; a significant prognosticator for patients undergoing curative resection for hepatocellular carcinoma. J Surg Oncol,2005,92(4):331-336.
70 Cheah PL, Looi LM, Nazarina AR, Mun KS, Goh KL. Association of Ki67 with raised transaminases in hepatocellular carcinoma. Malays J Pathol,2008,30(2):103-107.
71 Cui J, Dong BW, Liang P, Yu XL, Yu DJ. Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. World J Gastroenterol,2004,10(10):1533-1536.
72 Parkin MD,Bray F,Ferlay J,and Pisani P. Global Cancer Statistics.2002 CA-Caner J Clin,2005,55(74-108).
73 Upik Anderiani Miskad SS.Hirotaka Kato,Hiroshi Yokozaki.Expression of PRL-3 phosphatase in human gastric carcinomas:close correlation with invasion and metastasis. Pathobiology.71(4),2004.176-184.
74 Parker BS,Argani E Cook BP,Liangfeng H,Chartrand SD,Zhang M,Saha S,
Bardeili A,Jiang Y St.Martin TB,Nacht M,Teicher BA,Klinger KW,Sukumar S,and Madden SL. Alterations in vascular gene expression in invasive breast carcinoma. Cancer Res, 2004,64(21):7857-7866.
75 Weidner N.Tumowral vaseularity as a prognostic factor in cancer patients:the evidence continues to grow. J Pathol.184,1998.119-122.
76 Yoshiji H,Kutiyama s,Yoshii J,et al.Synergistic effect of basic fibroblast growth factor and vascular endothelial growth factor in mutine hepatocellar carcinoma. Hepatology, 2002,35:834-842.
77 Tischer E,Mitchell R, Hartman T,el al.The human gene for vascular endothelial growth factor. J Bio Chem,1991,226(11947-11954).
78 Kinlura H,Nakajima T,Kagawa K,et al.Angiogenesesen hepatoeellular carcinoma sevaluated by CD34 immunohistoehemistry. Liver,1998,18:14-19.
79 Ashton AW, Yokota R, John G, Zhao S, Suadicani SO, Spray DC, Ware JA. Inhibition of endothelial cell migration, intercellular communication, and vascular tube formation by thromboxane A(2).J BIOL CHEM,1999,274:35562-35570.
80 Soeda S, Kozako T, Iwata K, Shimeno H. Oversulfated fucoidan inhibits the basic fibroblast growth factor-induced tube formation by human umbilical vein endothelial cells:its possible mechanism of action. BIOCHIM BIOPHYS ACTA,2000,1497:127-134.
81贺国安,罗进贤,张添元,陈宏,孙慈煌.改进的鸡胚绒毛尿囊膜技术.中山大学学报(自然科学版),2003,42(2)：126-128.
82 He GA, Luo JX, Zhang TY, Hu ZS, Wang FY. The C-terminal domain of canstatin suppresses in vivo tumor growth associated with proliferation of endothelial cells. Biochem Biophys Res Commun,2004,318(2):354-360.
83 Weider N. Current pathologic methods for measuring intratumoral microvessel density within beast carcinoma and other solid tumors. Brest cancer Res Treat,1995,36:169-180.
84 Jain PK. Tumor angiogenesis and accessibility:role of vascular endothelial growth factor. Semin Oncol,2002,29(6 suppl-16):3-9.
85 Folkman J. Tumor angiogenesis:therapeutic implications. J N En-gl J Med, 1971,285:1182-1186.
86 Ferrara NH,Houek K,Jakeman L,et al.Molecular and biological properties of the vascular endothelial growth faetor family of proteins. J Endoer Rev,1992,13(1):18-32.
87 Gould J, Aramburo C, Capdevielle M, Scanes CG. Angiogenic activity of anterior pituitary tissue and growth hormone on the chick embryo chorio-allantoic membrane:a novel action of GH. Life Sci,1995,56(8):587-594.
88 Fraser RA, Ellis EM, Stalker AL. Experimental angiogenesis in the chorio-allantoic membrane. Bibl Anat.1979;(18):25-7. No abstract available,1979(18):25-27.
89 Griffoni C, Spisni E, Santi S, Riccio M, Guarnieri T, Tomasi V. Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo. Biochem Biophys Res Commun,2001,276(2):756-761.
90王世军,孙静,张栋.鸡胚尿囊膜血管生长的特点及观测方法.生物医学工程研究，2004,23(1)：3840.
91 Weidner N,Semple JP, Welch WR, et al. Tumor angiogenesis and metastasis:correlation in invasive breast carcinoma. Engl J Med,1991,324(1-8).
92 Gasparini G,Harris AL. Clinical importance of the determination of tumor angiogenesis in
breast cancer:much more than a new prognostic tool. J Clin Oncol,1995,13(765).
93 Maeda K, Chung YS,Takatsuks S, et al. Tumor angiogennesis and tumor cell proliferation as prognostic indicators in gastric carcinoma. Br J Cancer,1995,72:319.
94 Schlingenmann RO, Rietveld FJ, de Waal RM, et al.Leukocyte antigen CD34 is expressed by a subset of cultured endothelial cells and on endothelial abluminal microprocesses in the tumor stroma. Lab Invest.62,1990.690-696.
95 Cengiz Kirmaz, Kemal Ozbilgin, Hasan Yuksel, et al. Increased expression of angiogenic markers in patients with seasonal allergic rhinitis.. European cytokine network, 2004,15(4):317-322.
96 Martin L,holeombe C,Renshow C,et al.Standandising the counting technique of neovascularisafion in invasive breast cancer. Breast Cancer Res Treat,1996,37(supply):34-39.
97 Semela D,Dufour JF. Angiogenesis and hepatocellular carcinoma. J Hepatol, 2004,41:864-880.
98 An FQ,Matsuda M.Fujii H,et al.Expression of vascular endothelial growth factor in surgical speciments of hepatocellular carcinoma. Cancer Res Clin Oncol,2000,126(3):153-160.
99 Moon WS,Rhyukh KH,Kang MJ,et al. Overexpression of VEGF and angiopoietin 2:a key to high vascularity of hepatocellular carcinoma?.Mod Pathol,2003,16(6):552-557.
100 Yao DE,Wu XH,Zhu Y,et al.Quantitative analysis of vascular endothelial growth factor'microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4(2):220-226.
101 Ng IO,Poon RT,Lee JM,et al. Microvessel density,vascular endothelial growth factor and its receptors Fh-1 and Flk-1/KDR in hepatocellular carcinoma. Am J Clin Pathol, 2001,116(6):838-845.
102 Claudio PP, Russo G, Kumar CA,et al.pRb2/p138,vascular endothelial growth factor p27 (KIP1), and proliferating cell nuclear antigen expression in hepatocellular carcinoma:their clinical significance. Clin Cancer Res,2004,10(10):3509-3517.
103 Dvorak HF,Nagy JA,Feng D.et al.Vascular permeability factor vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Mivrobiol Imunol,1999,237(97-132).
104 Nor JE, Christensen J,Mooney DJ,et al.Vascular endothelial growth factor (VEGF)-mediated angiogenesis is associated with enhanced endotllelial cell survival and induction of bcl-2 expression. Am J Pathol,1999,154:375-384.
105 Gupta K, Kshirsagar S,Li W,et al.VEGF prevents appoptasis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling. Exp Cell Res, 1999,247:495-504.
106李江,王文亮,刘斌.人类肝细胞肝癌的细胞凋亡和癌组织内血管形成的调控.世界华人消化杂志,1998,6(12)：1057-1060.
107 Tseng PL, Tai MH, Huang CC, et al. Overexpression of VEGF is associated with positive p53 immunostaining in hepatocellular carcinoma (HCC) and adverse outcome of HCC patients. J Surg Oncol,2008,98:349-357.
108 Wada H, Nagano H, Yamamoto H, et al. Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma:importance of angiopoietin-2 and hypoxia-induced factor-1 alpha. Liver Int,2006,26:414-423.
109 Yao DF, Wu XH, Zhu Y, et al. Quantitative analysis of vascular endothelial growth factor,
microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4:220-226.
110 Huang GW, Yang LY, Lu WQ. Expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor in hepatocellular carcinoma:impact on neovascularization and survival. World J Gastroenterol,2005,11:1705-1708.
111 Kanda M, Nomoto S, Nishikawa Y, et al.Correlations of the expression of vascular endothelial growth factor B and its isoforms in hepatocellular carcinoma with clinico-pathological parameters. J Surg Oncol,2008,98:190-196.
112 Cui J, Dong BW, Liang P, Yu XL, Yu DJ.Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. World J Gastroenterol,2004,10:1533-1536.
113 Guo RP, Zhong C, Shi M, et al. Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma. J Cancer Res Clin Oncol.132,2006. 547-555.
114 Sheen IS, Jeng KS, Shih SC, et al.Clinical significance of the expression of isoform 165 vascular endothelial growth factor mRNA in noncancerous liver remnants of patients with hepatocellular carcinoma. World J Gastroenterol,2005,11:187-192.
115 Wang ZL, Liang P, Dong BW, Yu XL, Yu de J. Prognostic factors and recurrence of small hepatocellular carcinoma after hepatic resection or microwave ablation:a retrospective study. J Gastrointest Surg,2008,12:327-337.
116 Egeblad M,Werb. New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer,2002,2(3):161-174.
117 Longerich T,Breuhahn K, Odenthal M, et al.Factors of transfor ming growth factor beta signalling are co-regulated in human hepatocellular carcinoma. Virchows Arch, 2004,445(6):589-596.
118 Guo RP, Zhong C, Shi M, Zhang CQ, Wei W, Zhang YQ, Li JQ. Expression and clinical impact of vascular endothelial growth factor and matrix metalloproteinase-2 in hepatocellular carcinoma. Zhonghua Zhong Liu Za Zhi,2006,28(4):285-288.
119 Lee S, Jilani SM, Nikolova GV, et al. Processing of VEGF-A by matrix metalloproteinases regulates bioavailability and vascular patterning in tumors. J Cell Biol.169(4),2005.681-691.
120 Garzetti GG, Ciavattini A, Lucarini G, et al. Expression of vascular endothelial growth factor related to 72-kilodalton metalloproteinase immunostaining in patients with serous ovarian tumors. Cancer,1999,85(10):2219-2225.
121 Rooprai HK, Rucklidge GJ, Panou C, et al. The effects of exogenous growth factors on matrix metalloproteinase secretion by human brain tumour cells. Br J Cancer,2000,82(1):52-55.
122 Yoshiji Y,Kuriyama S,Noguchi R,et al.Angiopoietin 2 displays a vascular endothelial growth factor dependent synergistic effect in hepatocellular carcinoma development in mice. Gut, 2005,54(12):1768-1775.
123 Zhou XD.Recurrence and metastasis of hepatocellular carcinoma:progress and prospects. Hepatobiliary Pancreat Dis Int,2002,1(1):35-41.
124 Guo RP, Zhong C, Shi M, Zhang CQ, Wei W, Zhang YQ, Li JQ. Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma. J Cancer Res Clin Oncol,2006,132(9):547-555.
1 Zhu AX, Blaszkowsky LS, Ryan DP, et al. Phase II study of gemcitabine and oxaliplatin in combination with bevacizumab in patients with advanced hepatocellular carcinoma. J Clin Oncol,2006,24:1898-1903.
2 Hurwitz H, Fehrenbacher L, Novotny W, et al.Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med,2004,350:2335-2342.
3 Kabbinavar FF, Schulz J, McCleod M,et al. Addition of bevacizumab to bolus fluorouracil and leucovorin in firstline metastatic colorectal cancer:results of a randomized phase Ⅱ trial. J Clin Oncol,2005,23:3697-3705.
4 Sandler A, Gray R, Perry MC, et al. Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer. N Engl J Med,2006,355:2542-2550.
5 Ramaswamy B, Elias AD, Kelbick NT, et al.Phase Ⅱ trial of bevacizumab in combination with weekly docetaxel in metastatic breast cancer patients. Clin Cancer Res,2006,12:3124-3129.
6 Kindler HL, Friberg G, Singh DA, et al. Phase Ⅱ trial of bevacizumab plus gemcitabine in patients with advanced pancreatic cancer. J Clin Oncol,2005,23:8033-8040.
7 Sun W, Haller DG, Mykulowycz K, et al. Combination of capecitabine, oxaliplatin with bevacizumab in treatment of advanced hepatocellular carcinoma (HCC):a phase Ⅱ study. J Clin Oncol,2007,25 (supply):4574.
8 Llovet JM,Ricci S, Mazzaferro V, et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med, 2008,359:378-390.
9 Gruenwald V, Wilkens L, Gebel M, et al. A phase Ⅱ openlabel study of cetuximab in unresectable hepatocellular carcinoma. J Clin Oncol,2006,24((18supply)):14079.
10 O'Dwyer P, Giantonio B, Levy DE, et al. Gefitinib in advanced unresectable hepatocellular carcinoma: results from the Eastern Cooperative Oncology Group's study E1203. J Clin Oncol,2006,24(supply):4143.
11 Abou-Alfa GK, Schwartz L, Ricci S, et al. Phase Ⅱ study of sorafenib in patients with advanced hepatocellular carcinoma. J Clin Oncol,2006,24:4293-4300.
12 Siegel AB, Cohen El, Ocean A, et al. Phase Ⅱ trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma. J Clin Oncol,2008,26:2992-2998.
13 Thomas MB, Chadha R, Glover K, et al. Phase 2 study of erlotinib in patients with unresectable hepatocellular carcinoma. Cancer,2007,110:1059-1067.
14 Philip PA, Mahoney MR, Allmer C, et al. Phase Ⅱ study of erlotinib (OSI-774) in patients with advanced hepatocellular cancer. J Clin Oncol,2005,23:6657-6663.
15 Zhu AX, Stuart K, Blaszkowsky LS, et al. Phase 2 study of cetuximab in patients with advanced hepatocellular carcinoma. Cancer,2007,110:581-589.
16 Cheng AL, Kang YK, Chen Z, et al. Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma:a phase Ⅲ randomised, double-blind, placebo-controlled trial. Lancet Oncol,2009,10:25-34.
17 World Health Organization. Mortality database. WHO Statistical Information System. Available at: http://www.who.int/whosis Accessed March 2009.
18 Parkin DM, Pisani P, Ferlay J. Estimates of the worldwide incidence of 25 major cancers in 1990. Int J Cancer,1999,80:827-841.
19 Montalto G, Cervello M, Giannitrapani L, Dantona F, Terranova A, Castagnetta LA. Epidemiology, risk
factors, and natural history of hepatocellular carcinoma. Ann N Y Acad Sci,2002,963:13-20.
20 Sherman M.Hepatocellular carcinoma:epidemiology, risk factors, and screening. Semin Liver Dis, 2005,25:143-154.
21 Parkin DM,Bray F, Ferlay J, Pisani P. Global cancer statistics,2002.CA Cancer J Clin,2005,55:74-108.
22 Yeh CT, Chen TC, Chang ML, Hsu CW, Yeh TS, Lee WC, Huang SF, Tsai CC. Identification of NV-F virus DNA in hepatocellular carcinoma. J Med Virol,2007,79:92-96.
23汤钊献.肝癌临床研究现状,2008.
24徐克.我国肝细胞癌TACE治疗的现状与进展.2008.
25 Di Bisceglie AM.Epidemiology and clinical presentation of hepatocellular carcinoma. J Vasc Interv Radiol, 2002,13:S169-S171.
26 El-Serag HB, Mason AC. Rising incidence of hepatocellular carcinoma in the United States. N Engl J Med, 1999,340:745-750.
27 Taylor-Robinson SD, Thomas HC, Arora S, Hargreaves S. Increased mortality from liver cancer in England and Wales is not related to hepatitis C. BMJ,1999,319:640.
28 Khan SA, Taylor-Robinson SD, Toledano MB, Beck A, Elliott P, Thomas HC. Changing international trends in mortality rates for liver, biliary and pancreatic tumours. J Hepatol,2002,37:806-813.
29 American Cancer Society. Cancer Facts and Figures. Atlanta, GA:American Cancer Society; 2008.
30 El-Serag HB. Epidemiology of hepatocellular carcinoma in USA. Hepatol Res,2007,37 Suppl 2:S88-S94.
31 Parikh S, Hyman D. Hepatocellular cancer:a guide for the internist. Am J Med,2007,120:194-202.
32 Bosch FX, Ribes J, Diaz M, Cleries R. Primary liver cancer:worldwide incidence and trends. Gastroenterology,2004,127:S5-S16.
33 Bosch FX, Ribes J, Borras J. Epidemiology of primary liver cancer. Semin Liver Dis.19,1999.271-285.
34 Gupta S, Bent S, Kohlwes J. Test characteristics of alphafetoprotein for detecting hepatocellular carcinoma in patients with hepatitis C. A systematic review and critical analysis. Ann Intern Med,2003,139:46-50.
35 Ding X,Park YN,Taltavuil TC,Thung SN,Jin XM,Jin Y Trung NS,Edamoto Y Sata T,and Abe K. K,Geographic characterization of hepatitis virus infections,genotyping of hepatitis B virus,and p53 mutation in hepatocellular carcinoma analyzed by in situ detection of viral genomes from carcinoma tissues: Comparison among six different countries.pn.J.Infect.Dis.56(1),2003.12-18.
36 Carr BI.Hepatocellular carcinoma:Current management and future trends. Gastroenterology, 2004,127(5):S218-s224.
37 Poon RTP,Fan ST,Lo CM,Ng IOL,Liu CL, Lam CM,and Wong J. J,Improving survival results after resection of hepatocellular carcinoma:A prospective study of 377 patients over 10 years. Ann.Surg, 2001,234(1):63-70.
38 Tang ZY,Ye SL,Liu YK,Qin LX,Sun HC,Ye QH,Wang L,Zhou J,Qiu SJ,Li Y Ji XN,Liu H, Xia JL, Wu ZQ,Fan J,Ma ZC,Zhou XD,Lin ZY and Liu KD. A decade'S studies on metastasis of hepatocellular carcinoma.Cancer Res.Clin.Oncol,2004,130(4):187-196.
39 Lok ASF and Lai CL. a-Fetoprotein monitoring in chinese patients with chronic hepatitis B virus infection:role in the early detection of hepatocellular carcinoma. Hepatology,1989,9(1):110-115.
40 Sherman M,Pelteldan KM,and Lee C. Screening for hepatocellular carcinoma in chronic carriers of hepatitis B virus:incidence and prevalence of hepatocellular carcinoma in a North-American urban population. Hepatology,1995,22(2):432-438.
41 Folkman J.Tumor angiogenesis:therapeutic implications. N Engl J Med,1971,285(21):1182.
42 Folkman J. Angiogenesis in cancer,vascular,rheumatoid and other disease. Nat Med,1995,1(1):27-31.
43 Hanahan D,Folkman J.Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis.
Cell,1996,86(3):353.
44 Mignatti P and Rifkin DB.Biology and biochemistry of proteinases in tumor invasion.Physiol Rev, 1993,73(1):161-165.
45 Thaloor D,Singh A K, Sidhu GS,et al.Inhibition of angiogen2 ic differentiation of human umbilical vein endot helial cells by curcumin.Cell Growt h Differ,1998,9(4):305.
46 Fidler IJ, Yano S, Zhang RD, Fujimaki T, Bucana CD. The seed and soil hypothesis:vascularisation and brain metastases. Lancet Oncol.3,2002.53-57.
47 Senger DR, Galli SJ, Dvorak AM, Perruzzi CA, Harvey VS, Dvorak HF.Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science,1983,219:983-985.
48 Medina J,Arroyo A,Sanchez-Madrid F'and Moreno-Otero &.Angiogenesis in chronic inflammatory liver disease. Hepatology,2004,39(5):1185-1195.
49 Risau W. Mechanisms ofangiogenesis. Nature,1997,386(6626):671-674.
50 Dupuy E,Hainaud P,Villemain A,Bodevin-Phedre E,Brouland皿Pascale B, and Tobelem G. Tumoral angiogenesis and tissue factor expression during hepatocellular carcinoma progression in a transgenic mouse model. J Hepatoil,2003,38(6):793-802.
51 Foiberg凡Hendrix MJC,and Maniotis AJ. Vasculogenic mimicry and tumor angiogenesis. Am.J.Pathol, 2000,156(2):361-381.
52 Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors. Nat Med,2003,9(669-676).
53 Davis-Smyth T,Presta L G,Ferrara N. Mapping the charged residues in the second irmunoglobulin-1 ike domain of vascular endothelial growth factor/placenta growth factor receptor FLT-1 required for binding and structural stability. J Biol Chem,1998,273:3216-3222.
54 Kroll J,Waltenberger J. The vascular endothelial growth factor receptor KDR activates multiple cells. J Biol Chem,1997,272(51):32521-32527.
55 Waltenberger J,Claesson Welsh L, Siegahn A,et al.Different signal transduction properties of KDR and FLT-1,two receptor of vascular endothelial growth factor. J Biol Chem,1994,269(43):26988-26995.
56 Joukov V.Pajusola K,Kaipainen A,et al.A novel vascular endothelial growth factor. VEGF-C is a ligand for the FLT-4 and KDR receptot tyrosine kinase. EMBO J,1996,15(2):290-298.
57 Sulpice E, Plouet J, Berge M, Allanic D, Tobelem G, Merkulova-Rainon T. Neuropilin-1 and neuropilin-2 act as coreceptors, potentiating proangiogenic activity. Blood,2008,111:2036-2045.
58 Dvorak HF,Nagy JA,Feng D,et al.Vascular permeability factor vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Mivrobiol Imunol, 1999,237(97-132).
59 Nor JE, Christensen J,Mooney DJ,et al.Vascular endothelial growth factor (VEGF)-mediated angiogenesis is associated with enhanced endotllelial cell survival and induction of bcl-2 expression. Am J Pathol, 1999,154:375-384.
60 Gupta K,Kshirsagar S,Li W,et al.VEGF prevents appoptasis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling. Exp Cell Res,1999,247:495-504.
61 Ravi R, Mookergee B, Bhujwalla Z, et al. Regulation of tumor angio-genesis by p53 induced degradation of hypoxia-inducible factor 1α. Genes Dev.14,2000.34-44.
62 Brodsky SV,Mendelev N,Melamed M, et al. Vascular density and VEGF exp ression in hepatic lessions. J Gastrointestin Liver Dis.16(4),2007.373-377.
63 Yao DF,Wu XH, Zhu Y, et al. Quantitative analysis of vascular endothelial growth factor,microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4(2):220-226.
64 Kanematsu M, Osada S, Amaoka N, et al.Expression of vascular endothelial growth factor in hepatocellular carcinoma and the surrounding liver:correlation with angiographically assisted CT. AJR Am J Roentgenol, 2004,183:1585-1593.
65 Wang B, Gao ZQ, Yan X. Correlative study of angiogenesis and dynamic contrast-enhanced magnetic resonance imaging features of hepatocellular carcinoma. Acta Radiol,2005,46:353-358.
66 Tseng PL, Tai MH, Huang CC, et al. Overexpression of VEGF is associated with positive p53 immunostaining in hepatocellular carcinoma (HCC) and adverse outcome of HCC patients. J Surg Oncol, 2008,98:349-357.
67 Wada H, Nagano H, Yamamoto H, et al. Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma:importance of angiopoietin-2 and hypoxia-induced factor-1 alpha. Liver Int,2006,26:414-423.
68 Iavarone M,Lampertico P, Iannuzzi F, et al.Increased expression of vascular endothelial growth factor in small hepatocellular carcinoma. J Viral Hepat,2007,16:133-139.
69 Brodsky SV, Mendelev N, Melamed M, Ramaswamy G. Vascular density and VEGF expression in hepatic lesions. Gastrointestin Liver Dis,2007,16:373-377.
70 Yao DF, Wu XH, Zhu Y, et al. Quantitative analysis of vascular endothelial growth factor, microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4:220-226.
71 Huang GW, Yang LY, Lu WQ. Expression of hypoxia-inducible factor lalpha and vascular endothelial growth factor in hepatocellular carcinoma:impact on neovascularization and survival. World J Gastroenterol, 2005,11:1705-1708.
72 Kanda M,Nomoto S, Nishikawa Y, et al. Correlations of the expression of vascular endothelial growth factor B and its isoforms in hepatocellular carcinoma with clinico-pathological parameters. J Surg Oncol, 2008,98:190-196.
73 Claudio PP, Russo G, Kumar CA, et al. pRb2/p130, vascular endothelial growth factor, p27(KIP1),and proliferating cell nuclear antigen expression in hepatocellular carcinoma:their clinical significance. Clin Cancer Res,2004,10:3509-3517.
74 Cui J, Dong BW, Liang P, Yu XL, Yu DJ. Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. World J Gastroenterol, 2004,10:1533-1536.
75 Guo RP, Zhong C, Shi M, et al. Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma. J Cancer Res Clin Oncol.132,2006.547-555.
76 Sheen IS, Jeng KS, Shih SC, et al. Clinical significance of the expression of isoform 165 vascular endothelial growth factor mRNA in noncancerous liver remnants of patients with hepatocellular carcinoma. World J Gastroenterol,2005,11:187-192.
77 Wang ZL, Liang P, Dong BW, Yu XL, Yu de J. Prognostic factors and recurrence of small hepatocellular carcinoma after hepatic resection or microwave ablation:a retrospective study. J Gastrointest Surg, 2008,12:327-337.
78 Poon RT, Lau CP, Cheung ST, Yu WC, Fan ST. Quantitative correlation of serum levels and tumor expression of vascular endothelial growth factor in patients with hepatocellular carcinoma. Cancer Res, 2003,63(3121-3126).
79 Tseng CS, Lo HW, Chen PH, Chuang WL, Juan CC, Ker CG. Clinical significance of plasma D-dimer levels and serum VEGF levels in patients with hepatocellular carcinoma. Hepato-gastroenterology, 2004,51:1454-1458.
80 Sultan AS, S-el G, Hessien M, et al. Molecularmarkers of hepatitis C virus-related hepatocellular carcinoma. Cancer Biol Ther,2006,5(6):623-629.
81 Zhao J, Hu J, Cai J, Yang X, Yang Z. Vascular endothelial growth factor expression in serum of patients with hepatocellular carcinoma. Chin Med J (Engl),2003,116:772-776.
82 Poon RT, Ho JW, Tong CS, Lau C, Ng IO, Fan ST. Prognostic significance of serum vascular endothelial growth factor and endostatin in patients with hepatocellular carcinoma. Br J Surg,2004,91:1354-1360.
83 el-Houseini ME, Mohammed MS, Elshemey WM,Hussein TD, Desouky OS, Elsayed AA. Enhanced detection of hepatocellular carcinoma. Cancer Control,2005,12(4):248-253.
84 Iguchi H, Yokota M, Fukutomi M,et al. A possible role of VEGF in osteolytic bone metastasis of hepatocellular carcinoma. J Exp Clin Cancer Res,2002,21(3):309-313.
85 Poon RT, Lau CP, Cheung ST, Yu WC, Fan ST. Quantitative correlation of serum levels and tumor expression of vascular endothelial growth factor in patients with hepatocellular carcinoma. Cancer Res, 2003,63:3121-3126.
86 Banks RE, Forbes MA, Kinsey SE, et al.Release of the angiogenic cytokine vascular endothelial growth factor (VEGF) from platelets:significance for VEGF measurements and cancer biology. Br J Cancer, 1998,77:956-964.
87 Webb NJ, Bottomley MJ, Watson CJ, Brenchley PE. Vascular endothelial growth factor (VEGF) is released from platelets during blood clotting:implications for measurement of circulating VEGF levels in clinical disease. Clin Sci(Lond),1998,94:395-404.
88 Jeng KS, Sheen IS, Wang YC, et al.Prognostic significance of preoperative circulating vascular endothelial growth factor messenger RNA expression in resectable hepatocellularcarcinoma:a prospective study. World J Gastroenterol,2004,10:643-648.
89 Omenn GS. Strategies for plasma proteomic profiling of cancers. Proteomics,2006,6:5662-5673.
90 Siegel AB, Cohen El, Ocean A, et al. Phase Ⅱ trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma. J Clin Oncol,2008,26:2992-2998.
91 Thomas MB, Morris JS, Chadha R, et al. Phase Ⅱ trial of the combination of bevacizumab and erlotinib in patients who have advanced hepatocellular carcinoma. J Clin Oncol,2009,27:843-850.
92 Bruix J, Sherman M, Llovet JM, et al. Clinical management of hepatocellular carcinoma. Conclusions of the Barcelona-2000 EASL conference. European Association for the Study of the Liver. J Hepatol, 2001,35:421-430.
2汤钊猷.肝癌临床研究现状,2008.
3徐克.我国肝细胞癌TACE治疗的现状与进展.2008.
4 Endo K.Ueda T,Ueyama J,Ohta T, Terada T. lmmunoreactive E-cadherin,aipha-catenin, beta-catenin,and gamma-catenin proteins in hepatocellular carcinoma: Relationships with tumor grade,clinicopathologic parameters,and patients'survival. Hum.Pathol, 2003,31(5):558-565.
5 Ulmer SC. Hepatocellular carcinoma:a concise guide to its status and management. Postgrad Med,2000,107:117-124.
6 Shaw RJ,Cantley LC. Ras,PI(3)K and mTOR signalling controls tumour cell growth. Nature. 441(7092),2006.424-430.
7 Michor F,lwasa Y Nowak MA.Dynamics of cancer progression. Nat Rev Cancer, 2004,4(3):197-205.
8 Mangues R Symmans WF, Lu S,Schwartz S,Pellicer A. Activated N-ras oncogene and N-ras proto-oncogene act through the same pathway for in vivo tumorigenesis..Oncogene, 1996,13(5):1053-1063.
9 Prochownik EV. c-Myc:linking transformation and genomic instability. Curr Mol Med, 2008,8(6):446-458.
10 Hainaut P,Hollstein M. p53 and human cancer:the fa st ten thousand mutations. Adv Cancer Res,2000,77:81-137.
11 Minamoto T,Buschmann T,Habelhah H,Matusevich E,Taham H,Boerresen-Dale AL,Harris C,Sidransky D,Ronai Z. Distinct pattern of p53 phosphorylation in human tumors. Oncogene,2001,20(26):3341-3347.
12 Wang SP,Zhou HJ,Chen XP, Ren GY,Ruan XX,Zhang Y,Zhang RL,Chen J. Loss of expression of Kruppel-like factor 6 in primary hepatocellular carcinoma and hepatoma cell lines. J Exp Clin Cancer Res,2007,26(1):117-124.
13 Natoli G,Iamfi A, Costanzo A,et al.Resistance to Fas-mediated apoptosis in human hepatoma cells. Oneogene.11,1991.1157-1164.
14 Gulbins E,Jelde A,Ferlinz K, et al. Physiology of apoptosis. Am J Physiol Renal Physiol, 2000,279:605-615.
15 Carrington PE, Sandu C, Wei Y, et al. The structure of FADD and its mode of interaction with procaspase-8.Mol Cell,2006,22(5):599-610.
16 Tmmer T, Devries EG, DE Jong S. Fas receptor-mediated apoptosis:a clinical application. J Pathol,2003,15(11):983-992.
17 Donepudi M,Mac Sweeney A,Briand C,et al.Insights into the regulatory mechanism for caspase-8 activation. Mol Cell,2003,11:543-549.
18 Rodier F,Berlr R, Bossolasco M,et al. Polyomavims large T-antigen protects mouse cells
from Fas-,TNF-a and toxol-induced apoptosis. Oncogen,2000,19:6261-6270.
19 Liu A, Han X, Zhang G, Zhao X, Zheng G. Effects of cartilage polysaccharide on apoptosis of human hepatoma BEL-7402 cells and murine H22 hepatocarcinoma. Int J Food Sci Nutr, 2009,31:1-12.
20张晨,李柏,吕书勤,李勇,苏永华,凌昌全.蜂毒素对人肝癌细胞线粒体膜蛋白7A6和凋亡相关基因产物Fas及其配体FasL表达的影响.中西医结合学报,2007,5(5)：559-563.
21 Wu LQ, Zhang WJ, Niu JX, Ye LY,Yang ZH,Grau GE and Lou JN. Phenotypic and functional differences between human liver Barlcer endothelial cells and liver sinusoidal endothelial cells. J Vase Res,2008,45:78-86.
22 Nelson A, Fingleton B,Rothenberg ML,and Matrisian LM. Matrix metalloproteinases: Biologic activity and clinical implications. J.Ciin.Oneol,2000,18(5):1135-1149.
23 Livak KJ,Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods,2001,25(4):402-408.
24 S.Shen, W. Li, M. Ouyang, Z. Wu, Q. Lin, L. Xie. Identification of two marine fungi and evaluation of their antivirus and antitumor activities. Wei Sheng Wu Xue Bao, 2009,49:1240-1246.
25 U.W. Hawas, M. Shaaban, K.A. Shaaban, M. Speitling, A. Maier, G. Kelter, H.H.Fiebig, M. Meiners, E. Helmke, H. Laatsch. Mansouramycins A-D, cytotoxic isoquinolinequinones from a marine streptomycete. J Nat Prod,2009,72:2120-2124.
26 P.J. Schupp, C. Kohlert-Schupp, S. Whitefield, A. Engemann, S. Rohde, T. Hemscheidt, J.M. Pezzuto, T.P. Kondratyuk, E.J. Park, L. Marler, B. Rostama, A.D. Wright. Cancer chemopreventive and anticancer evaluation of extracts and fractions from marine macro-and microorganisms collected from Twilight Zone waters around Guam. Nat Prod Commun, 2009,4:1717-1728.
27 Li GY, Liu JZ, Yu XM, Chen SF, Zhang B, Zhang WF, et al.Effect of a seashell protein Haishengsu on cell growth and expression of apoptosis genes in leukemia K562 cells. Clin Invest Med,2008,31:E218-E221.
28 Ji-Zhu Liu, Shou-Guo Chen,Zhang Bin,Chun-Bo Wang, Guang-Yao Li,Le-Xin Wang. Antitumor effect of the seashell protein Haishengsu on Ehrlich ascites tumor:an experimental study. J Nat Med,2009,63:459-462.
29张成喜,李光耀,韩春山,叶小坤,陈洪英,周红娟,陈守国..泥蚶提取物-海生素制剂治疗中、晚期非小细胞肺癌和晚期肾癌的临床观察.中国海洋药物.24(4),2005.55-58.
30李光耀,张成喜,辛艳春,周红娟,张新,李花兰,王春波,陈守国..海生素制剂对非小细胞肺癌化疗患者生存质量的影响.中国海洋药物,2005,24(6)：50-51.
31 Li GY, Yu XM, Zhang HW, Zhang B, Wang CB, Xin YC, Yang CZ, et al. Haishengsu as an adjunct therapy to conventional chemotherapy in patients with non-small cell lung cancer:a pilot randomized and placebo-controlled clinical trial. Complement Ther Med,2009,17(1):51-55.
32 Lambert C,Landau A M,Desbarats J. Fas-beyond death:a regenerative role for Fas in the nervous system. Apoptosis,2003,8(551-562).
33 Li-Weber M,Krammer PH. Function and regulation of the CD95(APO-1/Fas) ligand in the immune system. Semin Immunol,2003,15:145-157.
34 Wajant H,Pfizenmaier K, Scheurich P. Tumor necrosis factor signaling. Cell Death Differ, 2003,10:45-63.
35 Anastassiou G,Coupland S E,Stang A,et al.Expression of Fas and Fas ligand in uveal
melanoma:biological implication and prognostic value. J Pathol,2001,194:466-472.
36 Silvestris F,Tucci M,Cafforio P,et al.Fas-L-up-regulation by highly malignant myelomant plasma cells:role in the pathogenesis of anemia and disease progression. Blood, 2001,97:1155-1164.
37 Korkolopoulou P,Goudopoulou A,Vout sinas G,et al.c-FLIP expression in bladder urothelial carcinomas:it's role in resistance to Fas-mediated apoptosis and clinicopat hologic correlations. U-rology,2004,63(1198-1204).
38 Strand S,Hofmann W J,Hug H,et al.Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing tumor cells-a mechanism of immunle evasion. N at Med,1996,2:1361-1366.
39 Nagao M,Nakajima Y,Hisanaga M,et al.The alteration of Fas receptor and ligand system in hepatocellular carcinomas:how do hepatoma cells escape from the host immune surveillance in vivo. J Hepatol,1999,30:413-421.
40 Y Cao, RW Ji, D Davidson, et al. Kringle domains of human angiostatin characterization of the anti-proliferative activity on endothelial cells. J Biol Chem,1996,271:29461-29467.
41 T Mosmann. Rapid colorimetric assay for cellular growth and survival:Application to proliferation and cytotoxicity assays. Journal of Immunological Methods,1983,65(1-2):55-63.
42 Y Cao, A Chen, SSA An, et al. Kringle 5 of Plasminogen is a Novel Inhibitor of Endothelial Cell Growth. J Biol Chem,1997,272:22924-22928.
43 T. Mitsuishi, Y. Iwabu, K. Tokunaga, T. Sata, T. Kaneko, K. Ohara, I. Ohsawa, F. Oda, Y. Yamada, S. Kawana, K. Ozaki, M. Nakatake, O. Yamada. Combined analysis of cell growth and apoptosis-regulating proteins in HPVs associated anogenital tumors. BMC Cancer,2010,10:118.
44 E.L. Kim, R. Wustenberg, A. Rubsam, C. Schmitz-Salue, G. Warnecke, E.M. Bucker, N. Pettkus, D. Speidel, V. Rohde, W. Schulz-Schaeffer, W. Deppert, A. Giese. Chloroquine activates the p53 pathway and induces apoptosis in human glioma cells. Neuro Oncol,2010,12:389-400.
45 S.Senthivinayagam, P. Mishra, S.K. Paramasivam, S.Yallapragada, M. Chatterjee, L. Wong, A. Rana, B.Rana. Caspase-mediated cleavage of beta-catenin precedes drug-induced apoptosis in resistant cancer cells. J Biol Chem,2009,284:13577-13588.
46 Kerr JF,Wyllie AH,Cullrie AR. Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br. J.Cancer,1972,26:239-257.
47 Wyllie AH,Kerr JF,Currie AR. Cell death in the significance of apoptosis. Int. Rev.Cytol,1980,68:251-306.
48 Dcbatin K M,Krammer P H. Death reception in chemotherapy and Cancer. Oncogene, 2004,23:2950-2966.
49 Ashkenazi A,Dixit V M. Death Receptors:Signaling and Modulation. Science, 1998,281:1305-1308.
50 Q. Chen, J.H. Zhang, J.T. Chen, Z.J. Shi, Y.J. Deng. MTT assay for detecting osteosarcoma cell apoptosis. Nan Fang Yi Ke Da Xue Xue Bao,2009,29:1899-1901.
51 Wesche-Soldato D E,Swan R Z,Chung C S,et al. The apoptotic pathway as a therapeutic target in sepsis. Curt Drug Targets,2007,8:493-500.
52 Everett H,McFadden G. Apoptosis:An innate immune response to virus infection. Trends Microbiol,1997,7:160-165.
53 Los M,Burek C J,Stroh C,et al.Anticancer drugs of tomorrow:Apoptotic pathways as targets for drug design. Drug Discov Today,2003,8:67-77.
54 Thompson C B. Apoptosis in the pathogenesis and treatment of disease. Science,
1995,267:1456-1462.
55 Thorbum J,Moore F,Rao A,et al.Selective Inaction of a Fas-associated Death Domain Protein (FADD)-dependent Apoptosis and Autophagy Pathway in Immon Epithelial Cells. Cells Mol Biol Cell,2005,16:1189-1199.
56 Bamhart B C,Alappat E C,Peter M E. The CD95 type Ⅰ/type Ⅱ model.Semin Immunol, 2003,15:185-193.
57 Kulms D, Zeise E, Poppelmann B, Schwarz T. DNA damage, death receptor activation and reactive oxygen species contribute to ultraviolet radiation-induced apoptosis in an essential and independent way. Oncogene,2002,196:5844-5851.
58 D.A.Martin,R.M.Siegel,L.Zheng,M.J.Lenardo. Membrane oligomerization and cleavage activates the c aspase-8(FLICE/MACHal)death signal. J.Biol.Chem,1998,274:4345-4349.
59 Bamhart B C,Alappat E C,Peter M E. The CD95 type Ⅰ/type Ⅱ model. Semin Immunol, 2003,15:185-193.
60 Boatfight K M,Salvesen G S. Mechanisms of caspase activation. Curt Opin Cell Biol, 2003,15:725-731.
61 Luo X,Budihardjo I,Zou H,Slaughter C,Wang X. Bid,a Bcl-2 interacting protein,mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell,1998,94:481-490.
62 Galvan V, Chen S, Lu D, et al. Caspase cleavage of members of the amyloid precursor family of proteins. J Neurochem.
63汪谦,主编.现代医学实验方法.北京：民卫生出版社：北京,1998.1094.
64 Canis M,Botchorishvili R,Wattiez A,et al. Tumor growth and dissemination after laparotomy and C02 pneumoperitoneum:a rat ovarian,cancer model.Obstet Gynecol, 1998,92(1):104-108.
65周际昌.实用肿瘤内科学.京：人民卫生出版社,1998.82-94.
66 Hu L,Hofmann J,Lu Y,et al.Inhibition of phosphatidylinositol 3'-kinase increases efficacy of pacl itaxel in vitro and in vivo ovarian cancer models. Cancer Res,2002,62(4):1087-1092.
67魏泓.医学实验动物学.成都：四川科学出版社,1998.302-303.
68司徒振强,吴军正.细胞培养.北京：世界图书出版社,1996.193.
69 Morinaga S, Ishiwa N, Noguchi Y, Yamamoto Y, Rino Y, Imada T, Takanashi Y, Akaike M, Sugimasa Y, Takemiya S. Growth index, assessed with Ki-67 and ssDNA labeling; a significant prognosticator for patients undergoing curative resection for hepatocellular carcinoma. J Surg Oncol,2005,92(4):331-336.
70 Cheah PL, Looi LM, Nazarina AR, Mun KS, Goh KL. Association of Ki67 with raised transaminases in hepatocellular carcinoma. Malays J Pathol,2008,30(2):103-107.
71 Cui J, Dong BW, Liang P, Yu XL, Yu DJ. Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. World J Gastroenterol,2004,10(10):1533-1536.
72 Parkin MD,Bray F,Ferlay J,and Pisani P. Global Cancer Statistics.2002 CA-Caner J Clin,2005,55(74-108).
73 Upik Anderiani Miskad SS.Hirotaka Kato,Hiroshi Yokozaki.Expression of PRL-3 phosphatase in human gastric carcinomas:close correlation with invasion and metastasis. Pathobiology.71(4),2004.176-184.
74 Parker BS,Argani E Cook BP,Liangfeng H,Chartrand SD,Zhang M,Saha S,
Bardeili A,Jiang Y St.Martin TB,Nacht M,Teicher BA,Klinger KW,Sukumar S,and Madden SL. Alterations in vascular gene expression in invasive breast carcinoma. Cancer Res, 2004,64(21):7857-7866.
75 Weidner N.Tumowral vaseularity as a prognostic factor in cancer patients:the evidence continues to grow. J Pathol.184,1998.119-122.
76 Yoshiji H,Kutiyama s,Yoshii J,et al.Synergistic effect of basic fibroblast growth factor and vascular endothelial growth factor in mutine hepatocellar carcinoma. Hepatology, 2002,35:834-842.
77 Tischer E,Mitchell R, Hartman T,el al.The human gene for vascular endothelial growth factor. J Bio Chem,1991,226(11947-11954).
78 Kinlura H,Nakajima T,Kagawa K,et al.Angiogenesesen hepatoeellular carcinoma sevaluated by CD34 immunohistoehemistry. Liver,1998,18:14-19.
79 Ashton AW, Yokota R, John G, Zhao S, Suadicani SO, Spray DC, Ware JA. Inhibition of endothelial cell migration, intercellular communication, and vascular tube formation by thromboxane A(2).J BIOL CHEM,1999,274:35562-35570.
80 Soeda S, Kozako T, Iwata K, Shimeno H. Oversulfated fucoidan inhibits the basic fibroblast growth factor-induced tube formation by human umbilical vein endothelial cells:its possible mechanism of action. BIOCHIM BIOPHYS ACTA,2000,1497:127-134.
81贺国安,罗进贤,张添元,陈宏,孙慈煌.改进的鸡胚绒毛尿囊膜技术.中山大学学报(自然科学版),2003,42(2)：126-128.
82 He GA, Luo JX, Zhang TY, Hu ZS, Wang FY. The C-terminal domain of canstatin suppresses in vivo tumor growth associated with proliferation of endothelial cells. Biochem Biophys Res Commun,2004,318(2):354-360.
83 Weider N. Current pathologic methods for measuring intratumoral microvessel density within beast carcinoma and other solid tumors. Brest cancer Res Treat,1995,36:169-180.
84 Jain PK. Tumor angiogenesis and accessibility:role of vascular endothelial growth factor. Semin Oncol,2002,29(6 suppl-16):3-9.
85 Folkman J. Tumor angiogenesis:therapeutic implications. J N En-gl J Med, 1971,285:1182-1186.
86 Ferrara NH,Houek K,Jakeman L,et al.Molecular and biological properties of the vascular endothelial growth faetor family of proteins. J Endoer Rev,1992,13(1):18-32.
87 Gould J, Aramburo C, Capdevielle M, Scanes CG. Angiogenic activity of anterior pituitary tissue and growth hormone on the chick embryo chorio-allantoic membrane:a novel action of GH. Life Sci,1995,56(8):587-594.
88 Fraser RA, Ellis EM, Stalker AL. Experimental angiogenesis in the chorio-allantoic membrane. Bibl Anat.1979;(18):25-7. No abstract available,1979(18):25-27.
89 Griffoni C, Spisni E, Santi S, Riccio M, Guarnieri T, Tomasi V. Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo. Biochem Biophys Res Commun,2001,276(2):756-761.
90王世军,孙静,张栋.鸡胚尿囊膜血管生长的特点及观测方法.生物医学工程研究，2004,23(1)：3840.
91 Weidner N,Semple JP, Welch WR, et al. Tumor angiogenesis and metastasis:correlation in invasive breast carcinoma. Engl J Med,1991,324(1-8).
92 Gasparini G,Harris AL. Clinical importance of the determination of tumor angiogenesis in
breast cancer:much more than a new prognostic tool. J Clin Oncol,1995,13(765).
93 Maeda K, Chung YS,Takatsuks S, et al. Tumor angiogennesis and tumor cell proliferation as prognostic indicators in gastric carcinoma. Br J Cancer,1995,72:319.
94 Schlingenmann RO, Rietveld FJ, de Waal RM, et al.Leukocyte antigen CD34 is expressed by a subset of cultured endothelial cells and on endothelial abluminal microprocesses in the tumor stroma. Lab Invest.62,1990.690-696.
95 Cengiz Kirmaz, Kemal Ozbilgin, Hasan Yuksel, et al. Increased expression of angiogenic markers in patients with seasonal allergic rhinitis.. European cytokine network, 2004,15(4):317-322.
96 Martin L,holeombe C,Renshow C,et al.Standandising the counting technique of neovascularisafion in invasive breast cancer. Breast Cancer Res Treat,1996,37(supply):34-39.
97 Semela D,Dufour JF. Angiogenesis and hepatocellular carcinoma. J Hepatol, 2004,41:864-880.
98 An FQ,Matsuda M.Fujii H,et al.Expression of vascular endothelial growth factor in surgical speciments of hepatocellular carcinoma. Cancer Res Clin Oncol,2000,126(3):153-160.
99 Moon WS,Rhyukh KH,Kang MJ,et al. Overexpression of VEGF and angiopoietin 2:a key to high vascularity of hepatocellular carcinoma?.Mod Pathol,2003,16(6):552-557.
100 Yao DE,Wu XH,Zhu Y,et al.Quantitative analysis of vascular endothelial growth factor'microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4(2):220-226.
101 Ng IO,Poon RT,Lee JM,et al. Microvessel density,vascular endothelial growth factor and its receptors Fh-1 and Flk-1/KDR in hepatocellular carcinoma. Am J Clin Pathol, 2001,116(6):838-845.
102 Claudio PP, Russo G, Kumar CA,et al.pRb2/p138,vascular endothelial growth factor p27 (KIP1), and proliferating cell nuclear antigen expression in hepatocellular carcinoma:their clinical significance. Clin Cancer Res,2004,10(10):3509-3517.
103 Dvorak HF,Nagy JA,Feng D.et al.Vascular permeability factor vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Mivrobiol Imunol,1999,237(97-132).
104 Nor JE, Christensen J,Mooney DJ,et al.Vascular endothelial growth factor (VEGF)-mediated angiogenesis is associated with enhanced endotllelial cell survival and induction of bcl-2 expression. Am J Pathol,1999,154:375-384.
105 Gupta K, Kshirsagar S,Li W,et al.VEGF prevents appoptasis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling. Exp Cell Res, 1999,247:495-504.
106李江,王文亮,刘斌.人类肝细胞肝癌的细胞凋亡和癌组织内血管形成的调控.世界华人消化杂志,1998,6(12)：1057-1060.
107 Tseng PL, Tai MH, Huang CC, et al. Overexpression of VEGF is associated with positive p53 immunostaining in hepatocellular carcinoma (HCC) and adverse outcome of HCC patients. J Surg Oncol,2008,98:349-357.
108 Wada H, Nagano H, Yamamoto H, et al. Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma:importance of angiopoietin-2 and hypoxia-induced factor-1 alpha. Liver Int,2006,26:414-423.
109 Yao DF, Wu XH, Zhu Y, et al. Quantitative analysis of vascular endothelial growth factor,
microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4:220-226.
110 Huang GW, Yang LY, Lu WQ. Expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor in hepatocellular carcinoma:impact on neovascularization and survival. World J Gastroenterol,2005,11:1705-1708.
111 Kanda M, Nomoto S, Nishikawa Y, et al.Correlations of the expression of vascular endothelial growth factor B and its isoforms in hepatocellular carcinoma with clinico-pathological parameters. J Surg Oncol,2008,98:190-196.
112 Cui J, Dong BW, Liang P, Yu XL, Yu DJ.Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. World J Gastroenterol,2004,10:1533-1536.
113 Guo RP, Zhong C, Shi M, et al. Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma. J Cancer Res Clin Oncol.132,2006. 547-555.
114 Sheen IS, Jeng KS, Shih SC, et al.Clinical significance of the expression of isoform 165 vascular endothelial growth factor mRNA in noncancerous liver remnants of patients with hepatocellular carcinoma. World J Gastroenterol,2005,11:187-192.
115 Wang ZL, Liang P, Dong BW, Yu XL, Yu de J. Prognostic factors and recurrence of small hepatocellular carcinoma after hepatic resection or microwave ablation:a retrospective study. J Gastrointest Surg,2008,12:327-337.
116 Egeblad M,Werb. New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer,2002,2(3):161-174.
117 Longerich T,Breuhahn K, Odenthal M, et al.Factors of transfor ming growth factor beta signalling are co-regulated in human hepatocellular carcinoma. Virchows Arch, 2004,445(6):589-596.
118 Guo RP, Zhong C, Shi M, Zhang CQ, Wei W, Zhang YQ, Li JQ. Expression and clinical impact of vascular endothelial growth factor and matrix metalloproteinase-2 in hepatocellular carcinoma. Zhonghua Zhong Liu Za Zhi,2006,28(4):285-288.
119 Lee S, Jilani SM, Nikolova GV, et al. Processing of VEGF-A by matrix metalloproteinases regulates bioavailability and vascular patterning in tumors. J Cell Biol.169(4),2005.681-691.
120 Garzetti GG, Ciavattini A, Lucarini G, et al. Expression of vascular endothelial growth factor related to 72-kilodalton metalloproteinase immunostaining in patients with serous ovarian tumors. Cancer,1999,85(10):2219-2225.
121 Rooprai HK, Rucklidge GJ, Panou C, et al. The effects of exogenous growth factors on matrix metalloproteinase secretion by human brain tumour cells. Br J Cancer,2000,82(1):52-55.
122 Yoshiji Y,Kuriyama S,Noguchi R,et al.Angiopoietin 2 displays a vascular endothelial growth factor dependent synergistic effect in hepatocellular carcinoma development in mice. Gut, 2005,54(12):1768-1775.
123 Zhou XD.Recurrence and metastasis of hepatocellular carcinoma:progress and prospects. Hepatobiliary Pancreat Dis Int,2002,1(1):35-41.
124 Guo RP, Zhong C, Shi M, Zhang CQ, Wei W, Zhang YQ, Li JQ. Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma. J Cancer Res Clin Oncol,2006,132(9):547-555.
1 Zhu AX, Blaszkowsky LS, Ryan DP, et al. Phase II study of gemcitabine and oxaliplatin in combination with bevacizumab in patients with advanced hepatocellular carcinoma. J Clin Oncol,2006,24:1898-1903.
2 Hurwitz H, Fehrenbacher L, Novotny W, et al.Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med,2004,350:2335-2342.
3 Kabbinavar FF, Schulz J, McCleod M,et al. Addition of bevacizumab to bolus fluorouracil and leucovorin in firstline metastatic colorectal cancer:results of a randomized phase Ⅱ trial. J Clin Oncol,2005,23:3697-3705.
4 Sandler A, Gray R, Perry MC, et al. Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer. N Engl J Med,2006,355:2542-2550.
5 Ramaswamy B, Elias AD, Kelbick NT, et al.Phase Ⅱ trial of bevacizumab in combination with weekly docetaxel in metastatic breast cancer patients. Clin Cancer Res,2006,12:3124-3129.
6 Kindler HL, Friberg G, Singh DA, et al. Phase Ⅱ trial of bevacizumab plus gemcitabine in patients with advanced pancreatic cancer. J Clin Oncol,2005,23:8033-8040.
7 Sun W, Haller DG, Mykulowycz K, et al. Combination of capecitabine, oxaliplatin with bevacizumab in treatment of advanced hepatocellular carcinoma (HCC):a phase Ⅱ study. J Clin Oncol,2007,25 (supply):4574.
8 Llovet JM,Ricci S, Mazzaferro V, et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med, 2008,359:378-390.
9 Gruenwald V, Wilkens L, Gebel M, et al. A phase Ⅱ openlabel study of cetuximab in unresectable hepatocellular carcinoma. J Clin Oncol,2006,24((18supply)):14079.
10 O'Dwyer P, Giantonio B, Levy DE, et al. Gefitinib in advanced unresectable hepatocellular carcinoma: results from the Eastern Cooperative Oncology Group's study E1203. J Clin Oncol,2006,24(supply):4143.
11 Abou-Alfa GK, Schwartz L, Ricci S, et al. Phase Ⅱ study of sorafenib in patients with advanced hepatocellular carcinoma. J Clin Oncol,2006,24:4293-4300.
12 Siegel AB, Cohen El, Ocean A, et al. Phase Ⅱ trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma. J Clin Oncol,2008,26:2992-2998.
13 Thomas MB, Chadha R, Glover K, et al. Phase 2 study of erlotinib in patients with unresectable hepatocellular carcinoma. Cancer,2007,110:1059-1067.
14 Philip PA, Mahoney MR, Allmer C, et al. Phase Ⅱ study of erlotinib (OSI-774) in patients with advanced hepatocellular cancer. J Clin Oncol,2005,23:6657-6663.
15 Zhu AX, Stuart K, Blaszkowsky LS, et al. Phase 2 study of cetuximab in patients with advanced hepatocellular carcinoma. Cancer,2007,110:581-589.
16 Cheng AL, Kang YK, Chen Z, et al. Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma:a phase Ⅲ randomised, double-blind, placebo-controlled trial. Lancet Oncol,2009,10:25-34.
17 World Health Organization. Mortality database. WHO Statistical Information System. Available at: http://www.who.int/whosis Accessed March 2009.
18 Parkin DM, Pisani P, Ferlay J. Estimates of the worldwide incidence of 25 major cancers in 1990. Int J Cancer,1999,80:827-841.
19 Montalto G, Cervello M, Giannitrapani L, Dantona F, Terranova A, Castagnetta LA. Epidemiology, risk
factors, and natural history of hepatocellular carcinoma. Ann N Y Acad Sci,2002,963:13-20.
20 Sherman M.Hepatocellular carcinoma:epidemiology, risk factors, and screening. Semin Liver Dis, 2005,25:143-154.
21 Parkin DM,Bray F, Ferlay J, Pisani P. Global cancer statistics,2002.CA Cancer J Clin,2005,55:74-108.
22 Yeh CT, Chen TC, Chang ML, Hsu CW, Yeh TS, Lee WC, Huang SF, Tsai CC. Identification of NV-F virus DNA in hepatocellular carcinoma. J Med Virol,2007,79:92-96.
23汤钊献.肝癌临床研究现状,2008.
24徐克.我国肝细胞癌TACE治疗的现状与进展.2008.
25 Di Bisceglie AM.Epidemiology and clinical presentation of hepatocellular carcinoma. J Vasc Interv Radiol, 2002,13:S169-S171.
26 El-Serag HB, Mason AC. Rising incidence of hepatocellular carcinoma in the United States. N Engl J Med, 1999,340:745-750.
27 Taylor-Robinson SD, Thomas HC, Arora S, Hargreaves S. Increased mortality from liver cancer in England and Wales is not related to hepatitis C. BMJ,1999,319:640.
28 Khan SA, Taylor-Robinson SD, Toledano MB, Beck A, Elliott P, Thomas HC. Changing international trends in mortality rates for liver, biliary and pancreatic tumours. J Hepatol,2002,37:806-813.
29 American Cancer Society. Cancer Facts and Figures. Atlanta, GA:American Cancer Society; 2008.
30 El-Serag HB. Epidemiology of hepatocellular carcinoma in USA. Hepatol Res,2007,37 Suppl 2:S88-S94.
31 Parikh S, Hyman D. Hepatocellular cancer:a guide for the internist. Am J Med,2007,120:194-202.
32 Bosch FX, Ribes J, Diaz M, Cleries R. Primary liver cancer:worldwide incidence and trends. Gastroenterology,2004,127:S5-S16.
33 Bosch FX, Ribes J, Borras J. Epidemiology of primary liver cancer. Semin Liver Dis.19,1999.271-285.
34 Gupta S, Bent S, Kohlwes J. Test characteristics of alphafetoprotein for detecting hepatocellular carcinoma in patients with hepatitis C. A systematic review and critical analysis. Ann Intern Med,2003,139:46-50.
35 Ding X,Park YN,Taltavuil TC,Thung SN,Jin XM,Jin Y Trung NS,Edamoto Y Sata T,and Abe K. K,Geographic characterization of hepatitis virus infections,genotyping of hepatitis B virus,and p53 mutation in hepatocellular carcinoma analyzed by in situ detection of viral genomes from carcinoma tissues: Comparison among six different countries.pn.J.Infect.Dis.56(1),2003.12-18.
36 Carr BI.Hepatocellular carcinoma:Current management and future trends. Gastroenterology, 2004,127(5):S218-s224.
37 Poon RTP,Fan ST,Lo CM,Ng IOL,Liu CL, Lam CM,and Wong J. J,Improving survival results after resection of hepatocellular carcinoma:A prospective study of 377 patients over 10 years. Ann.Surg, 2001,234(1):63-70.
38 Tang ZY,Ye SL,Liu YK,Qin LX,Sun HC,Ye QH,Wang L,Zhou J,Qiu SJ,Li Y Ji XN,Liu H, Xia JL, Wu ZQ,Fan J,Ma ZC,Zhou XD,Lin ZY and Liu KD. A decade'S studies on metastasis of hepatocellular carcinoma.Cancer Res.Clin.Oncol,2004,130(4):187-196.
39 Lok ASF and Lai CL. a-Fetoprotein monitoring in chinese patients with chronic hepatitis B virus infection:role in the early detection of hepatocellular carcinoma. Hepatology,1989,9(1):110-115.
40 Sherman M,Pelteldan KM,and Lee C. Screening for hepatocellular carcinoma in chronic carriers of hepatitis B virus:incidence and prevalence of hepatocellular carcinoma in a North-American urban population. Hepatology,1995,22(2):432-438.
41 Folkman J.Tumor angiogenesis:therapeutic implications. N Engl J Med,1971,285(21):1182.
42 Folkman J. Angiogenesis in cancer,vascular,rheumatoid and other disease. Nat Med,1995,1(1):27-31.
43 Hanahan D,Folkman J.Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis.
Cell,1996,86(3):353.
44 Mignatti P and Rifkin DB.Biology and biochemistry of proteinases in tumor invasion.Physiol Rev, 1993,73(1):161-165.
45 Thaloor D,Singh A K, Sidhu GS,et al.Inhibition of angiogen2 ic differentiation of human umbilical vein endot helial cells by curcumin.Cell Growt h Differ,1998,9(4):305.
46 Fidler IJ, Yano S, Zhang RD, Fujimaki T, Bucana CD. The seed and soil hypothesis:vascularisation and brain metastases. Lancet Oncol.3,2002.53-57.
47 Senger DR, Galli SJ, Dvorak AM, Perruzzi CA, Harvey VS, Dvorak HF.Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science,1983,219:983-985.
48 Medina J,Arroyo A,Sanchez-Madrid F'and Moreno-Otero &.Angiogenesis in chronic inflammatory liver disease. Hepatology,2004,39(5):1185-1195.
49 Risau W. Mechanisms ofangiogenesis. Nature,1997,386(6626):671-674.
50 Dupuy E,Hainaud P,Villemain A,Bodevin-Phedre E,Brouland皿Pascale B, and Tobelem G. Tumoral angiogenesis and tissue factor expression during hepatocellular carcinoma progression in a transgenic mouse model. J Hepatoil,2003,38(6):793-802.
51 Foiberg凡Hendrix MJC,and Maniotis AJ. Vasculogenic mimicry and tumor angiogenesis. Am.J.Pathol, 2000,156(2):361-381.
52 Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors. Nat Med,2003,9(669-676).
53 Davis-Smyth T,Presta L G,Ferrara N. Mapping the charged residues in the second irmunoglobulin-1 ike domain of vascular endothelial growth factor/placenta growth factor receptor FLT-1 required for binding and structural stability. J Biol Chem,1998,273:3216-3222.
54 Kroll J,Waltenberger J. The vascular endothelial growth factor receptor KDR activates multiple cells. J Biol Chem,1997,272(51):32521-32527.
55 Waltenberger J,Claesson Welsh L, Siegahn A,et al.Different signal transduction properties of KDR and FLT-1,two receptor of vascular endothelial growth factor. J Biol Chem,1994,269(43):26988-26995.
56 Joukov V.Pajusola K,Kaipainen A,et al.A novel vascular endothelial growth factor. VEGF-C is a ligand for the FLT-4 and KDR receptot tyrosine kinase. EMBO J,1996,15(2):290-298.
57 Sulpice E, Plouet J, Berge M, Allanic D, Tobelem G, Merkulova-Rainon T. Neuropilin-1 and neuropilin-2 act as coreceptors, potentiating proangiogenic activity. Blood,2008,111:2036-2045.
58 Dvorak HF,Nagy JA,Feng D,et al.Vascular permeability factor vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Mivrobiol Imunol, 1999,237(97-132).
59 Nor JE, Christensen J,Mooney DJ,et al.Vascular endothelial growth factor (VEGF)-mediated angiogenesis is associated with enhanced endotllelial cell survival and induction of bcl-2 expression. Am J Pathol, 1999,154:375-384.
60 Gupta K,Kshirsagar S,Li W,et al.VEGF prevents appoptasis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling. Exp Cell Res,1999,247:495-504.
61 Ravi R, Mookergee B, Bhujwalla Z, et al. Regulation of tumor angio-genesis by p53 induced degradation of hypoxia-inducible factor 1α. Genes Dev.14,2000.34-44.
62 Brodsky SV,Mendelev N,Melamed M, et al. Vascular density and VEGF exp ression in hepatic lessions. J Gastrointestin Liver Dis.16(4),2007.373-377.
63 Yao DF,Wu XH, Zhu Y, et al. Quantitative analysis of vascular endothelial growth factor,microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4(2):220-226.
64 Kanematsu M, Osada S, Amaoka N, et al.Expression of vascular endothelial growth factor in hepatocellular carcinoma and the surrounding liver:correlation with angiographically assisted CT. AJR Am J Roentgenol, 2004,183:1585-1593.
65 Wang B, Gao ZQ, Yan X. Correlative study of angiogenesis and dynamic contrast-enhanced magnetic resonance imaging features of hepatocellular carcinoma. Acta Radiol,2005,46:353-358.
66 Tseng PL, Tai MH, Huang CC, et al. Overexpression of VEGF is associated with positive p53 immunostaining in hepatocellular carcinoma (HCC) and adverse outcome of HCC patients. J Surg Oncol, 2008,98:349-357.
67 Wada H, Nagano H, Yamamoto H, et al. Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma:importance of angiopoietin-2 and hypoxia-induced factor-1 alpha. Liver Int,2006,26:414-423.
68 Iavarone M,Lampertico P, Iannuzzi F, et al.Increased expression of vascular endothelial growth factor in small hepatocellular carcinoma. J Viral Hepat,2007,16:133-139.
69 Brodsky SV, Mendelev N, Melamed M, Ramaswamy G. Vascular density and VEGF expression in hepatic lesions. Gastrointestin Liver Dis,2007,16:373-377.
70 Yao DF, Wu XH, Zhu Y, et al. Quantitative analysis of vascular endothelial growth factor, microvascular density and their clinicopathologic features in human hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int,2005,4:220-226.
71 Huang GW, Yang LY, Lu WQ. Expression of hypoxia-inducible factor lalpha and vascular endothelial growth factor in hepatocellular carcinoma:impact on neovascularization and survival. World J Gastroenterol, 2005,11:1705-1708.
72 Kanda M,Nomoto S, Nishikawa Y, et al. Correlations of the expression of vascular endothelial growth factor B and its isoforms in hepatocellular carcinoma with clinico-pathological parameters. J Surg Oncol, 2008,98:190-196.
73 Claudio PP, Russo G, Kumar CA, et al. pRb2/p130, vascular endothelial growth factor, p27(KIP1),and proliferating cell nuclear antigen expression in hepatocellular carcinoma:their clinical significance. Clin Cancer Res,2004,10:3509-3517.
74 Cui J, Dong BW, Liang P, Yu XL, Yu DJ. Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. World J Gastroenterol, 2004,10:1533-1536.
75 Guo RP, Zhong C, Shi M, et al. Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma. J Cancer Res Clin Oncol.132,2006.547-555.
76 Sheen IS, Jeng KS, Shih SC, et al. Clinical significance of the expression of isoform 165 vascular endothelial growth factor mRNA in noncancerous liver remnants of patients with hepatocellular carcinoma. World J Gastroenterol,2005,11:187-192.
77 Wang ZL, Liang P, Dong BW, Yu XL, Yu de J. Prognostic factors and recurrence of small hepatocellular carcinoma after hepatic resection or microwave ablation:a retrospective study. J Gastrointest Surg, 2008,12:327-337.
78 Poon RT, Lau CP, Cheung ST, Yu WC, Fan ST. Quantitative correlation of serum levels and tumor expression of vascular endothelial growth factor in patients with hepatocellular carcinoma. Cancer Res, 2003,63(3121-3126).
79 Tseng CS, Lo HW, Chen PH, Chuang WL, Juan CC, Ker CG. Clinical significance of plasma D-dimer levels and serum VEGF levels in patients with hepatocellular carcinoma. Hepato-gastroenterology, 2004,51:1454-1458.
80 Sultan AS, S-el G, Hessien M, et al. Molecularmarkers of hepatitis C virus-related hepatocellular carcinoma. Cancer Biol Ther,2006,5(6):623-629.
81 Zhao J, Hu J, Cai J, Yang X, Yang Z. Vascular endothelial growth factor expression in serum of patients with hepatocellular carcinoma. Chin Med J (Engl),2003,116:772-776.
82 Poon RT, Ho JW, Tong CS, Lau C, Ng IO, Fan ST. Prognostic significance of serum vascular endothelial growth factor and endostatin in patients with hepatocellular carcinoma. Br J Surg,2004,91:1354-1360.
83 el-Houseini ME, Mohammed MS, Elshemey WM,Hussein TD, Desouky OS, Elsayed AA. Enhanced detection of hepatocellular carcinoma. Cancer Control,2005,12(4):248-253.
84 Iguchi H, Yokota M, Fukutomi M,et al. A possible role of VEGF in osteolytic bone metastasis of hepatocellular carcinoma. J Exp Clin Cancer Res,2002,21(3):309-313.
85 Poon RT, Lau CP, Cheung ST, Yu WC, Fan ST. Quantitative correlation of serum levels and tumor expression of vascular endothelial growth factor in patients with hepatocellular carcinoma. Cancer Res, 2003,63:3121-3126.
86 Banks RE, Forbes MA, Kinsey SE, et al.Release of the angiogenic cytokine vascular endothelial growth factor (VEGF) from platelets:significance for VEGF measurements and cancer biology. Br J Cancer, 1998,77:956-964.
87 Webb NJ, Bottomley MJ, Watson CJ, Brenchley PE. Vascular endothelial growth factor (VEGF) is released from platelets during blood clotting:implications for measurement of circulating VEGF levels in clinical disease. Clin Sci(Lond),1998,94:395-404.
88 Jeng KS, Sheen IS, Wang YC, et al.Prognostic significance of preoperative circulating vascular endothelial growth factor messenger RNA expression in resectable hepatocellularcarcinoma:a prospective study. World J Gastroenterol,2004,10:643-648.
89 Omenn GS. Strategies for plasma proteomic profiling of cancers. Proteomics,2006,6:5662-5673.
90 Siegel AB, Cohen El, Ocean A, et al. Phase Ⅱ trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma. J Clin Oncol,2008,26:2992-2998.
91 Thomas MB, Morris JS, Chadha R, et al. Phase Ⅱ trial of the combination of bevacizumab and erlotinib in patients who have advanced hepatocellular carcinoma. J Clin Oncol,2009,27:843-850.
92 Bruix J, Sherman M, Llovet JM, et al. Clinical management of hepatocellular carcinoma. Conclusions of the Barcelona-2000 EASL conference. European Association for the Study of the Liver. J Hepatol, 2001,35:421-430.