肝癌复发的生物联合干预及基因谱表达调控的实验研究
|
摘要
提高人类原发性肝癌的疗效仍是医学界面临的难题。本研究利用生物治疗探索干预人原发性肝癌复发的治疗方案。基于细胞分子应答技术研究生物联合干预对肝癌细胞分化、细胞周期调控,癌组织血管抑制和免疫调控因子及基因表达谱的变化。应用基因芯片检测了人肝癌细胞BEL-7402细胞株的生物信息,体内、外分别使用生长抑素8肽、生长抑素8肽联合IFN—α2b干扰素调控肝癌细胞的分子应答机制。在不同时间,实验条件下,通过相差显微镜,苔盼蓝拒染法,流式细胞仪,基因芯片等检测手段。我们发现BEL-7402细胞与正常肝细胞在基因表达谱上有明显差异,在BEL-7402细胞系阳性表达的基因有79个。与正常人肝细胞表达谱有表达差异的基因有31个,上调24个,下调7个。生长抑素8肽体外实验发现对人肝癌细胞的抗增殖作用是通过影响细胞分化、细胞周期分布实现。表达差异的基因主要与G、S周期调控有关。细胞凋亡不是主要的作用机制。裸小鼠背部皮下注射BEL-7402细胞悬液,建立人原发性肝癌裸小鼠皮下移植模型。根据不同分组皮下注射生长抑素和生长抑素联合干扰素进行干预。通过观察皮下肿瘤生长情况,病理学检查,原位末端标记流式细胞仪检测细胞凋亡,免疫组化,微血管密度检测,基因芯片检测基因表达谱的变化。研究结果显示单独使用生长抑素对抑制人肝癌细胞移植瘤的增殖有作用,而联合使用生长抑素加干扰素则表现出更明显的肿瘤抑制效应。我们认为生长抑素抗人肝癌细胞移植瘤的作用机制是影响细胞的周期分布、细胞分化以及干预肿瘤血管生成。生长抑素联合干扰素使用对肿瘤细胞影响在于影响细胞周期分布,抑制了肿瘤新生血管的生成并且有促进瘤细胞凋亡的作用。基因表达谱显示23个基因有表
To investigate molecular characters of BEL-7402 , it were studied under cDNA microarray. Then to observe the effects of Somatostatin with Interferon-alpha inhibited proliferation of BEL-7402 hepatocellular carcinoma(HCC) cell line and growth of HCC xenografts in nude mice. Finally , to evaluate security of administration of Somatostatin in HCC patients after resection of liver. We compared gene expression alteration between BEL-7402 cell line and normal liver tissue by cDNA microarray. Among 866 cancer related genes, there were 71 genes had differential expression in BEL-7402 cell line. 24 genes were up-regulated while 7 genes were down-regulated. In those genes, many differentially expressed genes were involved in cell division, membrane receptor and immunocompeten. Abnormal gene expression contribute to malignant cell divison. Cell survive rates and activities were detected by typan blue exclusion respectively, phase contrast microscopy , flowcytometer and cDNA microarray were used to detect the ability to inhibit the growth of BEL-7402 in vitro. It may be an important mechanism for Somatostatin inhibition of proliferative ability of BEL-7402 cells by inducing cell differentiation and modulating the cell cycle progress, rather than by killing the hepatocellular carcinoma cell or inducing apoptosis directly. Nude mice bearing xenografts of cancer cell line were treated with Somatostatin, Somatostatin and Interferon-alpha or saline control for 4 weeks after tumor implantation .The mechanism of Inhibition of Somatostatin on proliferative ability of tumor cells was observed by
measuring tumor volume, MVD were detected with immunohistochemistry in three groups. Cell cycle progress and apoptosis were detected by flowcytometer, gene expression profile was identified by cDNA microarrays. The ratio of cell in resting state (G0/G1) increased, MVD decreasing , apopotosis peak was observed, there were 23 genes had differential expression. 7 genes were up-regulated while 16 genes were down-regulated, specific genes involved in modulating the cell cycle progress , apoptosis cascade. These finding suggest that the inhibition on effects occur in the growth of HCC transplant tumor in nude mice were markedly improved by Somatostatin combined with Interferon-alpha when compared with Somatostatin alone. To administrate Somatostatin was security after resection of liever, it could enhance wound healing. These synergic results may be of potential therapeutic benefit to those patients with HCC.
To investigate molecular characters of BEL-7402 , it were studied under cDNA microarray. Then to observe the effects of Somatostatin with Interferon-alpha inhibited proliferation of BEL-7402 hepatocellular carcinoma(HCC) cell line and growth of HCC xenografts in nude mice. Finally , to evaluate security of administration of Somatostatin in HCC patients after resection of liver. We compared gene expression alteration between BEL-7402 cell line and normal liver tissue by cDNA microarray. Among 866 cancer related genes, there were 71 genes had differential expression in BEL-7402 cell line. 24 genes were up-regulated while 7 genes were down-regulated. In those genes, many differentially expressed genes were involved in cell division, membrane receptor and immunocompeten. Abnormal gene expression contribute to malignant cell divison. Cell survive rates and activities were detected by typan blue exclusion respectively, phase contrast microscopy , flowcytometer and cDNA microarray were used to detect the ability to inhibit the growth of BEL-7402 in vitro. It may be an important mechanism for Somatostatin inhibition of proliferative ability of BEL-7402 cells by inducing cell differentiation and modulating the cell cycle progress, rather than by killing the hepatocellular carcinoma cell or inducing apoptosis directly. Nude mice bearing xenografts of cancer cell line were treated with Somatostatin, Somatostatin and Interferon-alpha or saline control for 4 weeks after tumor implantation .The mechanism of Inhibition of Somatostatin on proliferative ability of tumor cells was observed by
measuring tumor volume, MVD were detected with immunohistochemistry in three groups. Cell cycle progress and apoptosis were detected by flowcytometer, gene expression profile was identified by cDNA microarrays. The ratio of cell in resting state (G0/G1) increased, MVD decreasing , apopotosis peak was observed, there were 23 genes had differential expression. 7 genes were up-regulated while 16 genes were down-regulated, specific genes involved in modulating the cell cycle progress , apoptosis cascade. These finding suggest that the inhibition on effects occur in the growth of HCC transplant tumor in nude mice were markedly improved by Somatostatin combined with Interferon-alpha when compared with Somatostatin alone. To administrate Somatostatin was security after resection of liever, it could enhance wound healing. These synergic results may be of potential therapeutic benefit to those patients with HCC.
引文
1 汤钊猷主编.现代肿瘤学.第二版.上海:上海医科大学出版社,2000.44:737
2 Blum, -H-E. Molecular therapy and prevention of hepatocellular carcinoma. Hepatobiliary-Pancreat-Dis-Int. 2003 Feb; 2(1): 11-22
3 汤钊猷,余业勤主编.原发性肝癌.第二版.上海科学技术出版社1999,10
4 Parkin DM. Global cancer statistics in the year 2000[J]. Lancet Oncol, 2001, 2(9):533～543.
5 Okuno,-K; Yasutomi,-M. Multidisciplinary treatment for liver metastasis using cytokines, Nippon-Geka-Gakkai-Zasshi. 2001 102(5): 403-8
6 陈惠藜等.分化诱导剂对人肝癌细胞亚细胞组分酪胺酸蛋白激酶的早期的影响.上海医科大学学报,1999,26(3):197～199
7 潘启超.新抗癌药物的进展.见:曹世龙主编.肿瘤学新理论与技术.上海:上海科学技术出版社,1999,407～403
8 Yamada Y, kubato A,,Kagimoto S, et al. Identification of somatostatin receptor subtypes and an implication for the efficacy of somatostatin analogue SMS201-995 in treatment of human endocrine tumors. J Clin Inverst, 1994, 93:1321-1325.
9 Schindel, Jay L, Grosfeld. Hepatic resection enhances growth of residual intrahepatic and subcutaneous hepatoma, which is inhibited by octreotide. [J] Pediatr Surg, 1997,32(7):995～997.
10 Dimitroulopoulos,-D; Xinopoulos,-D; Tsamakidis,-K; Zisimopoulos,-A; Andriotis,-E; Markidou,-S; Panagiotakos,-D; Chrysohoou,-C; Bazinis,-A; Paraskevas,-E.The role of sandostatin LAR in treating patients with advanced hepatocellular cancer. Hepatogastroenterology. 2002 Sep-Oct; 49(47): 1245-50
11 刘建生,彭淑牖,吴育莲等.人类生长抑素受体mRAN在原发性肝癌中的表达.中华消化杂志2002,22:(11)683～684
12 Armstrong TD, Clements VK, Martin BK, et al. MHC-Ⅱ-transfected tumor cells present endogenous antigen and are potent dinducers of tumor-specific immumity. Pro Natl Acad Sci USA, 1997; 94(13):6886-6891
13 Fujii Y, Gonoi T, Yamada Y, Chihara K, Inagaki N, Seino S. Somatostatin receptor subtype SSTR2 mediates the inhibition of high-voltage-activated calcium channels by somatostatin and its analogue SMS 201-995.FEBS Lett. 1994 Nov 28; 355(2):117-20.
14 Ren SG, Ezzat S, Melmed S, et al. [J] Endocrinnology 1992, 131(5):2479-2481.
15 Elliott DE, Li J, Blum AM, Metwali A, Patel YC, Weinstock JV. SSTR2A is the dominant somatostatin receptor subtype expressed by inflammatory cells, is widely expressed and directly regulates T cell IFN-gamma release. Eur J Immunol. 1999 Aug;29(8):2454-63.
16 王建六,王一飞,孔红霞.肿瘤免疫研究进展,郑州,河南医科大学出版社,1996,139
17 Lin,-S-M; Lin,-C-J; Hsu,-C-W; Tai,-D-I; Sheen,-I-S; Lin,.D-Y; Liaw,-Y-F. Prospective randomized controlled study of interferon-alpha in preventing hepatocellular carcinoma recurrence after medical ablation therapy for primary tumors. Arch-Surg. 2001 May; 136(5): 569-75
18 O' Reilly MS, Holmgren L, Chen C, et al. Angiostatin induce and sustains dormancy of human primary tumors in mice. Nat Med. 1996; 2(6): 689-692
19 Rosenberg SA, Principles of cancer management: biologic therapy. In: Devita VT, Hellman S, Ronsenberg SA eds. Cancer: principle & practice of oncology, 5th ed. Philadelphia: Lippincott-Raven, 1997,349～373
20 Kwok,-P-C; Lam,-T-W; Lam,-P-W; Tang,-K-W; Chan,-S-C; Hwang,-J-S; Cheung,-M-T; Tang,-D-L; Chung,-T-K; Chia,-N-H; Wong,-W-K; Chan,-M-K; Lo,-H-Y; Lam,-W-M Randomized controlled trial to compare the dose of adjuvant therapy after curative resection of hepatocellular carcinoma. J-Gastroenterol-Hepatol. 2003 Apr; 18(4): 410-5
21 Shena M, Shalon D, Heller R, Chai A, Brown P O, Davis R W. Parallel human genome analysis microarraybased expression monitoring of 1000 genes, PNAS, USA, 1996, 93:10614～10619
22 Schena M, Shaion D, Dais R W. Brown P O. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. [J] Science.1995;270(5235):467-470.
23 Allan-k; Jordan-RC; et al. Overexpression of cyclin A and cyclin B proteins in astrocytomas. Arch Pathol Lab Med, 2000;124(2):216-20
24 徐路生,钱斌治等.肝癌相关基因表达差异分析.基础医学与临床,2002;22(3):223～227.
25 Burroughs KD, Oh J, Barrett JC, DiAugustine RP. Phosphatidylinositol 3-kinase and mek1/2 are necessary for insulin-like growth factor-I-induced vascular endothelial growth factor synthesis in prostate epithelial cells: a role for hypoxia-inducible factor-1? Mol Cancer Res. 2003 Feb;1(4):312-22
26 Shima Y, Kengaku M, Hirano T, Takeichi M, Uemura T. Regulation of dendritic maintenance and growth by a mammalian 7-pass transmembrane cadherin. Dev Cell. 2004 Aug;7(2):205-16.
27 章静波,林建银,杨恬主编.医学分子细胞生物学.北京:中国协和医科大学出版社,2002,431-433.
28 Shimizu T, Cai S, Tomioka H. Roles of reactive nitrogen intermediates and transforming growth factor-beta produced by immunosuppressive macrophages in the expression of suppressor activity against T cell proliferation induced by TCR stimulation. Cytokine. 2005 Apr 7;30(1):7-13
29 Michalides RJAM. Cell cycle regulators: mechanisms and their role in aetiology, prognosis, and treatment of cancer. [J] Clin Pathol, 1999;52:555-560
30 Chao Y, Shih YL, Chiu JH, Chau GY, Lui WY, Yang WK, Lee SD, Huang TS. Overexpression of cyclin A but not skp2 correlates with the tumor relapse of human hepatocellular carcinoma. Cancer Res, 1998;58: 985-990
31 黄振义.诱导分化治疗急性早期粒细胞性白血病的进展.见:曹世龙主编.肿瘤新技术与新理论.上海:上海科技教育出版社,1999,654～667
32 Sun Y, Li H Lin Z et al, phase I pharmacokinetic study of nemorubicin hydrochloride Methxymorpholino doxorubicin: (PNU-152243) administered with iodinated oil via hepatic artery (IH) to patients (PT) with unresectable hepatocellular carcinoma (HCC), proc ASCO 2003; 22;1448a
33 Kuiper RAI, Schellens JHm, Bdijham GH, et al. Clinical research on antiangiogenic therapy. Pharmacal Res. 1998; 37:1-16
34 Reubi JC, Laissue JA, Multiple actions of somatostatin in neoplastic disease. [J] Trends pharmacol Sci, 1995, 16(3):110-115.
35 吴细丕,钱林法主编.北京:中国医药科技出版社,2000,3
36 汤钊猷.裸鼠人体肝癌移植模型的研究Ⅰ:模型的建立及其血清学与形态学特征。[J]上海第一医学院学报,1982,(1):21～26
37 Sun FX, Tang ZY, Lui KD, et al. Establishment of a metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation of histologically intact tissues. [J] Int-J-Cancer, 1996, 66(2):239-43
38 Rock KL. A new foreign policy: MHC class I molecules monitor the outside world. Immunology Today,1996; 17(3):131-137
39 Kolenko VM, Uzzo RG, Bukowski R, et al. Caspase-dependent and independent pathways in cancer therapy. Apoptosis, 2000;5(1):17-20
40 Terrence G, Frey A, et al. Energy conversion mitochondria and respiratory chain expression. Trends in Biochemical Sciences, 2000,25:319
41. Folkman J: Angiogenesis: Therapeutic implication N. Engl J Med 1971; 285:1182-1186
42 Yamaguchi N, Anand-Apte B, Lee M, et al. Endostatin inhibits EGF-induced endothelial cell migration and tumor growth independently of zinc binding [J]. EMBO J, 1999;Aug16,18(16):4414-23
43 Eliceiri BP, Cheresh DA. The role of alphav integrins during angiogenesis: insights into potential mechanisms of action and clinical
44 Hunter T, Pines J. Cyclins and cancer Ⅱ: cyclin D and CDK inhibitors come of age. Cell, 1994; 79:573-582
45 朱斌,姚晓平,杨甲梅,丛文铭.原发性肝细胞癌中细胞周期G1期相关基因Cyclin D1的表达及其临床病理学意义.肝胆外科杂志,1999:7:214-216
46 Muller H, Lnkas J, Schneider A. Cyclin D1 expression is regulated by the retinoblastoma protein. Proc Natl Acad Sci
47 Chao Y, Shih YL, Chiu JH, Chau GY, Lui WY, Yang WK, Lee SD, Huang TS. Overexpression of cyclin A but not skp2 correlates with the tumor relapse of human hepatocellular carcinoma. Cancer Res 1998; 58:985-990
48 Schindel DT, Grosfeld JL. Hepatic resection enhances growth of residual intrahepatic and subcutaneous hepatoma, which is inhibited by octreotide.[J] Pediatr Surg. 1997 Jul;32(7):995-7; discussion 997-8
49 Hua Yun-Peng Huang Jie-Fu et al. Inhibition effects of Somatostatin on hepatocelullar carcinoma in vitro and vivo.[J] China Journal of Modern Medicine, 2003, 13(21) 14-17
50 Garcia JC, Escorsell A et al. Influence of pharmacological agents on portal hemodynamics:basis for its use in the treatment of portal hypertension.[J] Senin Liver Dis, 1999, 19(4):427-436.
51 Waddell BE, Calton W, Steinberg SR et al. The adverse effects of Octreotide on wound healing in rams. Am Surg, 1997, 63:446.
52 Turkaper AG, Deminer S, Sengul M et al. The adverse effects of Octreotide on healing of colonic anastomoses in rat. Surg Today, 1998, 28(3):279
53 Papalampros E, Felekours ES, Filis K, et al. Liver pathology and cell proloferation after Octreotide administration following partial hepatectomy in rats: an experimental study. [J] Dig Dis Sci. 2002, 47(9): 1953-1958.
54 Eileitson CD, Sherridan MA. Differential effects of somatostatin-14 and somatostatin-25 on carbohydrate and lipid metabolism in rainbow trout Onorhychus mykiss.[J] Gencomp Endocrinol.1993,62-70
55 Kamath PS, Wiesner RH, et al. A midel to predict survival in patients end-stsge liver disease. Hepatology, 2001, 33:468-470
56 Kouroumalis E, Skordilis P, Thermos K, et al. Drug therapy: Octreotide [J]. The New England Journal of Medicine, 1996, 334:246-254.
57 Kouroumalis E, Skordilis P, Thermos K, et al. Treatment of hepatocellular carcinoma with octreotide: a randomized controlled study [j]. Gut. 1998, 42(3):442-7.
58 Adachi E, Meada T, et al. Risk factors for interahepatic recurrence in human hepatocellular carcinoma. Gastronterology.1995,108:768-775.
59 Poon,-R-T; Fan,-S-T; Ng,-I-O; Wong,-J. Significance of resection margin in hepatectomy for hepatocellular carcinoma: A critical reappraisal. Ann-Surg. 2000 Apr; 231(4): 544-51
60 Tung-Ping-Poon; Fan,-S-T; Wong,-J. Risk factors, prevention, and management of postoperative recurrence after resection of hepatocellular carcinoma. Ann-Surg. 2000 Jul; 232(1): 10-24
61 Matsumura M, Niwa T, Ogura K, et al. AFPmRNA in blood as a marker to assess the efficacy of the treatments of hepatocellular carcinoma and predict its prognosis.Gastroenterology, 1996,110:1262-1264.
62 Zhou XD. Recurrence and metastasis of hepatocellular carcinoma: progress and prospects. Hepatobiliary Pancreat Dis Int. 2002 Feb;1(1):35-41.
63 Okuda K, Tanaka M, Nakayama T, et al. Clincopathologic comparison between resected hepatocellular carcinomas and recurrent tumors. Int Hepatology Commun. 1993.1(1):65
64 夏振龙.肝脏肿瘤病因学.见:黄洁夫主编.肝脏胆道肿瘤外科学.北京:人民卫生出版社,1999.25
65 Nagasue, -N; Ono, -T; Yamanoi, -A; Kohno,-H; EI-Assal, -O-N; Taniura,-H; Uchida,-M Prognostic factors and survival after hepatic resection for hepatocellular carcinoma without cirrhosis. Br-J-Surg. 2001 Apr; 88(4): 515-22
66 Hassoun,-Z; Gores,-G-J, What surgeons should know about viral hepatitis and hepatocellular carcinoma. Surg-Oncol-Clin-N-Am. 2003 Jan; 12(1): 1-11, ⅶ
67 Blum H. Molecular targets for prevention of hepatocellular carcinoma. Dig-Dis. 2002; 20(1):81-90
68 Lara-Pezzi E, Gomez-Gaviro MV, Galvez BG, et al. The hepatitis B virus X protein promotes tumor cell invasion by inducing membrane -type matrix metalloproteinase-1 and cyclooxygenase-2 expression. J Clin Invest. 2002 Dec; 110(12): 1831-8.
69 纪小龙、施作霖。诊断免疫组织化学.北京:军事医学科学出版社,1997;65
70 Osada S, Saji S, Kuno T. Clinical significance of combination study of apoptotic factors and proliferating cell nuclear antigen in estimating the prognosis of hepatocellular carcinoma. J Surg Oncol. 2004 Jan; 85(1):48-5.
71 Shi YZ, Hui AM, Takayama T, Li X, Cui X, Makuuchi M. Reduced p21(WAF1/CIP1) protein expression is predominantly related to altered p53 in hepatocellular carcinomas. Br J Cancer. 2000 Jul; 83(1):50-5
72 雷文东,张汝刚,阎水忠等,nm23基因在人肺癌中的表达及其与肺癌淋巴转移的关系。中华肿瘤杂志,1994;16:227
73 Yamaguchi A, Urano T, Goi T, et al. Expression of human nm23-H1 and nm23-H2 protein in hepatocellular carcinoma. [J]. Cancer,1994,73:2280.
74 Seki S, Kitada T, Kawada N, Sakaguchi H, Kadoya H, Nakatani K, Satake K. Estabilishment and characteristics of human hepatocelluar carcinoma cells with metastasis to lymph nodes. Hepatogastroenterology. 1999 Sep-Oct:46(29): 2812-7
75 Shimada M, Taguchi K, Hasegawa H, Gion T, Shirabe K, Tsuneyoshi M, Sugimachi K. Nm23-H1 expression in intrahepatic or extrahepatic metastases of hepatocellular carcinoma. Liver. 1998 Oct; 18(5):337-42
76 Nieto-Rodriguez A, Hernandez-Pando R, Kershenobich D, Rodriguez-Fragoso L. Expression of urokinase-type plasminogen activator in an experimental model of hepatocarcinoma. Toxicology. 2001 Mar 21;161(1-2):13-23
77 Maksan SM, Ariab PM, Ryschich E, Gebhard MM, Schmidt J. Immune escape mechanism: defective resting and stimulated leukocyte-endothelium interaction in hepatocellular carcinoma of the rat. Dig Dis Sci. 2004 May; 49 (5):859-65
78 Taketomi A, Takenaka K, Matsumata T, Shimada M, Higashi H, Shirabe K, Itasaka H, Adachi E, Maeda T, Sugimachi K. Circulating intercellular adhesion molecule-1 in patients with hepatocellular carcinoma before and after hepatic resection. Hepatogastroenterology. 1997 Mar-Apr;44(14):477-83
79 Kehrl JH, Roberts AB, Wakefield LM, et al. Transforming growth factor β is a important immunomodulatory protein for human B-lymphocytes. J Immunol.1986; 137:3855.
80 Shimizu S, Nishikawa Y, K uroda K, et al. Involvement of transforming growth factor β 1 in autocrine enhancement of gelatinase secretion by murine metastatic colon carcimoma cells. Cancer Res, 1996, 56:3366-3370
81 Bedossa P, Peltier E, Terris B, et al. Transforming growth factor betal (TGF-β_1) and TGF-β Receptor in normal, cirrhotic, and neoplastic human livers. Hepatology, 1995, 21 (3):760-766.
82 Matsumoto E, Nakatusukasa H, Nouso K, Kobayashi Y, Nakamura S, Suzuki M et al. Increased levels of tissue inhibitor of metalloproteinase-1 in human hepatocellular carcinoma. Liver int. 2004 Aug; 24(4):379-83
83 Miyoshi A, Kitajima Y, Sumi K, Soto K, Hagiwara A, Koga Y, Miyazaki K. Snail and SIP1 increase cancer invasion by upregulating MMP family in
84 ZHen HN, ZHang X, ZHang ZW, Bu XY, H. PZ, W ang BY. Expression of protooncogene c-erbB-1 and c-erbB-2 in human gliomas. Di-si Junyi Daxue Xuebao (J Fourth MiIMed U niv), 1999, 20 (2) 106-109
85 Hisaka T, Yano H haramaki M, et al. Expressions of epidernmal growth factor family and its receptor in hepatocellular carcimoma cell lines: relationship to cell proliferation [J]. Int J Oncol, 1999, 14:453
86 Arii S, Mori A, Uchida S, et al. Implication of vascular endothelial growth factor in the development and metastasis of human cancers.
87 Ueki T, fujimoto J, et al. Expression of hepatocyte growth factor and its receptor c-met proto-oncogene in hepatocellular carcinomal. [J]. Hepatology, 1997, 25(4):862-868.
2 Blum, -H-E. Molecular therapy and prevention of hepatocellular carcinoma. Hepatobiliary-Pancreat-Dis-Int. 2003 Feb; 2(1): 11-22
3 汤钊猷,余业勤主编.原发性肝癌.第二版.上海科学技术出版社1999,10
4 Parkin DM. Global cancer statistics in the year 2000[J]. Lancet Oncol, 2001, 2(9):533～543.
5 Okuno,-K; Yasutomi,-M. Multidisciplinary treatment for liver metastasis using cytokines, Nippon-Geka-Gakkai-Zasshi. 2001 102(5): 403-8
6 陈惠藜等.分化诱导剂对人肝癌细胞亚细胞组分酪胺酸蛋白激酶的早期的影响.上海医科大学学报,1999,26(3):197～199
7 潘启超.新抗癌药物的进展.见:曹世龙主编.肿瘤学新理论与技术.上海:上海科学技术出版社,1999,407～403
8 Yamada Y, kubato A,,Kagimoto S, et al. Identification of somatostatin receptor subtypes and an implication for the efficacy of somatostatin analogue SMS201-995 in treatment of human endocrine tumors. J Clin Inverst, 1994, 93:1321-1325.
9 Schindel, Jay L, Grosfeld. Hepatic resection enhances growth of residual intrahepatic and subcutaneous hepatoma, which is inhibited by octreotide. [J] Pediatr Surg, 1997,32(7):995～997.
10 Dimitroulopoulos,-D; Xinopoulos,-D; Tsamakidis,-K; Zisimopoulos,-A; Andriotis,-E; Markidou,-S; Panagiotakos,-D; Chrysohoou,-C; Bazinis,-A; Paraskevas,-E.The role of sandostatin LAR in treating patients with advanced hepatocellular cancer. Hepatogastroenterology. 2002 Sep-Oct; 49(47): 1245-50
11 刘建生,彭淑牖,吴育莲等.人类生长抑素受体mRAN在原发性肝癌中的表达.中华消化杂志2002,22:(11)683～684
12 Armstrong TD, Clements VK, Martin BK, et al. MHC-Ⅱ-transfected tumor cells present endogenous antigen and are potent dinducers of tumor-specific immumity. Pro Natl Acad Sci USA, 1997; 94(13):6886-6891
13 Fujii Y, Gonoi T, Yamada Y, Chihara K, Inagaki N, Seino S. Somatostatin receptor subtype SSTR2 mediates the inhibition of high-voltage-activated calcium channels by somatostatin and its analogue SMS 201-995.FEBS Lett. 1994 Nov 28; 355(2):117-20.
14 Ren SG, Ezzat S, Melmed S, et al. [J] Endocrinnology 1992, 131(5):2479-2481.
15 Elliott DE, Li J, Blum AM, Metwali A, Patel YC, Weinstock JV. SSTR2A is the dominant somatostatin receptor subtype expressed by inflammatory cells, is widely expressed and directly regulates T cell IFN-gamma release. Eur J Immunol. 1999 Aug;29(8):2454-63.
16 王建六,王一飞,孔红霞.肿瘤免疫研究进展,郑州,河南医科大学出版社,1996,139
17 Lin,-S-M; Lin,-C-J; Hsu,-C-W; Tai,-D-I; Sheen,-I-S; Lin,.D-Y; Liaw,-Y-F. Prospective randomized controlled study of interferon-alpha in preventing hepatocellular carcinoma recurrence after medical ablation therapy for primary tumors. Arch-Surg. 2001 May; 136(5): 569-75
18 O' Reilly MS, Holmgren L, Chen C, et al. Angiostatin induce and sustains dormancy of human primary tumors in mice. Nat Med. 1996; 2(6): 689-692
19 Rosenberg SA, Principles of cancer management: biologic therapy. In: Devita VT, Hellman S, Ronsenberg SA eds. Cancer: principle & practice of oncology, 5th ed. Philadelphia: Lippincott-Raven, 1997,349～373
20 Kwok,-P-C; Lam,-T-W; Lam,-P-W; Tang,-K-W; Chan,-S-C; Hwang,-J-S; Cheung,-M-T; Tang,-D-L; Chung,-T-K; Chia,-N-H; Wong,-W-K; Chan,-M-K; Lo,-H-Y; Lam,-W-M Randomized controlled trial to compare the dose of adjuvant therapy after curative resection of hepatocellular carcinoma. J-Gastroenterol-Hepatol. 2003 Apr; 18(4): 410-5
21 Shena M, Shalon D, Heller R, Chai A, Brown P O, Davis R W. Parallel human genome analysis microarraybased expression monitoring of 1000 genes, PNAS, USA, 1996, 93:10614～10619
22 Schena M, Shaion D, Dais R W. Brown P O. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. [J] Science.1995;270(5235):467-470.
23 Allan-k; Jordan-RC; et al. Overexpression of cyclin A and cyclin B proteins in astrocytomas. Arch Pathol Lab Med, 2000;124(2):216-20
24 徐路生,钱斌治等.肝癌相关基因表达差异分析.基础医学与临床,2002;22(3):223～227.
25 Burroughs KD, Oh J, Barrett JC, DiAugustine RP. Phosphatidylinositol 3-kinase and mek1/2 are necessary for insulin-like growth factor-I-induced vascular endothelial growth factor synthesis in prostate epithelial cells: a role for hypoxia-inducible factor-1? Mol Cancer Res. 2003 Feb;1(4):312-22
26 Shima Y, Kengaku M, Hirano T, Takeichi M, Uemura T. Regulation of dendritic maintenance and growth by a mammalian 7-pass transmembrane cadherin. Dev Cell. 2004 Aug;7(2):205-16.
27 章静波,林建银,杨恬主编.医学分子细胞生物学.北京:中国协和医科大学出版社,2002,431-433.
28 Shimizu T, Cai S, Tomioka H. Roles of reactive nitrogen intermediates and transforming growth factor-beta produced by immunosuppressive macrophages in the expression of suppressor activity against T cell proliferation induced by TCR stimulation. Cytokine. 2005 Apr 7;30(1):7-13
29 Michalides RJAM. Cell cycle regulators: mechanisms and their role in aetiology, prognosis, and treatment of cancer. [J] Clin Pathol, 1999;52:555-560
30 Chao Y, Shih YL, Chiu JH, Chau GY, Lui WY, Yang WK, Lee SD, Huang TS. Overexpression of cyclin A but not skp2 correlates with the tumor relapse of human hepatocellular carcinoma. Cancer Res, 1998;58: 985-990
31 黄振义.诱导分化治疗急性早期粒细胞性白血病的进展.见:曹世龙主编.肿瘤新技术与新理论.上海:上海科技教育出版社,1999,654～667
32 Sun Y, Li H Lin Z et al, phase I pharmacokinetic study of nemorubicin hydrochloride Methxymorpholino doxorubicin: (PNU-152243) administered with iodinated oil via hepatic artery (IH) to patients (PT) with unresectable hepatocellular carcinoma (HCC), proc ASCO 2003; 22;1448a
33 Kuiper RAI, Schellens JHm, Bdijham GH, et al. Clinical research on antiangiogenic therapy. Pharmacal Res. 1998; 37:1-16
34 Reubi JC, Laissue JA, Multiple actions of somatostatin in neoplastic disease. [J] Trends pharmacol Sci, 1995, 16(3):110-115.
35 吴细丕,钱林法主编.北京:中国医药科技出版社,2000,3
36 汤钊猷.裸鼠人体肝癌移植模型的研究Ⅰ:模型的建立及其血清学与形态学特征。[J]上海第一医学院学报,1982,(1):21～26
37 Sun FX, Tang ZY, Lui KD, et al. Establishment of a metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation of histologically intact tissues. [J] Int-J-Cancer, 1996, 66(2):239-43
38 Rock KL. A new foreign policy: MHC class I molecules monitor the outside world. Immunology Today,1996; 17(3):131-137
39 Kolenko VM, Uzzo RG, Bukowski R, et al. Caspase-dependent and independent pathways in cancer therapy. Apoptosis, 2000;5(1):17-20
40 Terrence G, Frey A, et al. Energy conversion mitochondria and respiratory chain expression. Trends in Biochemical Sciences, 2000,25:319
41. Folkman J: Angiogenesis: Therapeutic implication N. Engl J Med 1971; 285:1182-1186
42 Yamaguchi N, Anand-Apte B, Lee M, et al. Endostatin inhibits EGF-induced endothelial cell migration and tumor growth independently of zinc binding [J]. EMBO J, 1999;Aug16,18(16):4414-23
43 Eliceiri BP, Cheresh DA. The role of alphav integrins during angiogenesis: insights into potential mechanisms of action and clinical
44 Hunter T, Pines J. Cyclins and cancer Ⅱ: cyclin D and CDK inhibitors come of age. Cell, 1994; 79:573-582
45 朱斌,姚晓平,杨甲梅,丛文铭.原发性肝细胞癌中细胞周期G1期相关基因Cyclin D1的表达及其临床病理学意义.肝胆外科杂志,1999:7:214-216
46 Muller H, Lnkas J, Schneider A. Cyclin D1 expression is regulated by the retinoblastoma protein. Proc Natl Acad Sci
47 Chao Y, Shih YL, Chiu JH, Chau GY, Lui WY, Yang WK, Lee SD, Huang TS. Overexpression of cyclin A but not skp2 correlates with the tumor relapse of human hepatocellular carcinoma. Cancer Res 1998; 58:985-990
48 Schindel DT, Grosfeld JL. Hepatic resection enhances growth of residual intrahepatic and subcutaneous hepatoma, which is inhibited by octreotide.[J] Pediatr Surg. 1997 Jul;32(7):995-7; discussion 997-8
49 Hua Yun-Peng Huang Jie-Fu et al. Inhibition effects of Somatostatin on hepatocelullar carcinoma in vitro and vivo.[J] China Journal of Modern Medicine, 2003, 13(21) 14-17
50 Garcia JC, Escorsell A et al. Influence of pharmacological agents on portal hemodynamics:basis for its use in the treatment of portal hypertension.[J] Senin Liver Dis, 1999, 19(4):427-436.
51 Waddell BE, Calton W, Steinberg SR et al. The adverse effects of Octreotide on wound healing in rams. Am Surg, 1997, 63:446.
52 Turkaper AG, Deminer S, Sengul M et al. The adverse effects of Octreotide on healing of colonic anastomoses in rat. Surg Today, 1998, 28(3):279
53 Papalampros E, Felekours ES, Filis K, et al. Liver pathology and cell proloferation after Octreotide administration following partial hepatectomy in rats: an experimental study. [J] Dig Dis Sci. 2002, 47(9): 1953-1958.
54 Eileitson CD, Sherridan MA. Differential effects of somatostatin-14 and somatostatin-25 on carbohydrate and lipid metabolism in rainbow trout Onorhychus mykiss.[J] Gencomp Endocrinol.1993,62-70
55 Kamath PS, Wiesner RH, et al. A midel to predict survival in patients end-stsge liver disease. Hepatology, 2001, 33:468-470
56 Kouroumalis E, Skordilis P, Thermos K, et al. Drug therapy: Octreotide [J]. The New England Journal of Medicine, 1996, 334:246-254.
57 Kouroumalis E, Skordilis P, Thermos K, et al. Treatment of hepatocellular carcinoma with octreotide: a randomized controlled study [j]. Gut. 1998, 42(3):442-7.
58 Adachi E, Meada T, et al. Risk factors for interahepatic recurrence in human hepatocellular carcinoma. Gastronterology.1995,108:768-775.
59 Poon,-R-T; Fan,-S-T; Ng,-I-O; Wong,-J. Significance of resection margin in hepatectomy for hepatocellular carcinoma: A critical reappraisal. Ann-Surg. 2000 Apr; 231(4): 544-51
60 Tung-Ping-Poon; Fan,-S-T; Wong,-J. Risk factors, prevention, and management of postoperative recurrence after resection of hepatocellular carcinoma. Ann-Surg. 2000 Jul; 232(1): 10-24
61 Matsumura M, Niwa T, Ogura K, et al. AFPmRNA in blood as a marker to assess the efficacy of the treatments of hepatocellular carcinoma and predict its prognosis.Gastroenterology, 1996,110:1262-1264.
62 Zhou XD. Recurrence and metastasis of hepatocellular carcinoma: progress and prospects. Hepatobiliary Pancreat Dis Int. 2002 Feb;1(1):35-41.
63 Okuda K, Tanaka M, Nakayama T, et al. Clincopathologic comparison between resected hepatocellular carcinomas and recurrent tumors. Int Hepatology Commun. 1993.1(1):65
64 夏振龙.肝脏肿瘤病因学.见:黄洁夫主编.肝脏胆道肿瘤外科学.北京:人民卫生出版社,1999.25
65 Nagasue, -N; Ono, -T; Yamanoi, -A; Kohno,-H; EI-Assal, -O-N; Taniura,-H; Uchida,-M Prognostic factors and survival after hepatic resection for hepatocellular carcinoma without cirrhosis. Br-J-Surg. 2001 Apr; 88(4): 515-22
66 Hassoun,-Z; Gores,-G-J, What surgeons should know about viral hepatitis and hepatocellular carcinoma. Surg-Oncol-Clin-N-Am. 2003 Jan; 12(1): 1-11, ⅶ
67 Blum H. Molecular targets for prevention of hepatocellular carcinoma. Dig-Dis. 2002; 20(1):81-90
68 Lara-Pezzi E, Gomez-Gaviro MV, Galvez BG, et al. The hepatitis B virus X protein promotes tumor cell invasion by inducing membrane -type matrix metalloproteinase-1 and cyclooxygenase-2 expression. J Clin Invest. 2002 Dec; 110(12): 1831-8.
69 纪小龙、施作霖。诊断免疫组织化学.北京:军事医学科学出版社,1997;65
70 Osada S, Saji S, Kuno T. Clinical significance of combination study of apoptotic factors and proliferating cell nuclear antigen in estimating the prognosis of hepatocellular carcinoma. J Surg Oncol. 2004 Jan; 85(1):48-5.
71 Shi YZ, Hui AM, Takayama T, Li X, Cui X, Makuuchi M. Reduced p21(WAF1/CIP1) protein expression is predominantly related to altered p53 in hepatocellular carcinomas. Br J Cancer. 2000 Jul; 83(1):50-5
72 雷文东,张汝刚,阎水忠等,nm23基因在人肺癌中的表达及其与肺癌淋巴转移的关系。中华肿瘤杂志,1994;16:227
73 Yamaguchi A, Urano T, Goi T, et al. Expression of human nm23-H1 and nm23-H2 protein in hepatocellular carcinoma. [J]. Cancer,1994,73:2280.
74 Seki S, Kitada T, Kawada N, Sakaguchi H, Kadoya H, Nakatani K, Satake K. Estabilishment and characteristics of human hepatocelluar carcinoma cells with metastasis to lymph nodes. Hepatogastroenterology. 1999 Sep-Oct:46(29): 2812-7
75 Shimada M, Taguchi K, Hasegawa H, Gion T, Shirabe K, Tsuneyoshi M, Sugimachi K. Nm23-H1 expression in intrahepatic or extrahepatic metastases of hepatocellular carcinoma. Liver. 1998 Oct; 18(5):337-42
76 Nieto-Rodriguez A, Hernandez-Pando R, Kershenobich D, Rodriguez-Fragoso L. Expression of urokinase-type plasminogen activator in an experimental model of hepatocarcinoma. Toxicology. 2001 Mar 21;161(1-2):13-23
77 Maksan SM, Ariab PM, Ryschich E, Gebhard MM, Schmidt J. Immune escape mechanism: defective resting and stimulated leukocyte-endothelium interaction in hepatocellular carcinoma of the rat. Dig Dis Sci. 2004 May; 49 (5):859-65
78 Taketomi A, Takenaka K, Matsumata T, Shimada M, Higashi H, Shirabe K, Itasaka H, Adachi E, Maeda T, Sugimachi K. Circulating intercellular adhesion molecule-1 in patients with hepatocellular carcinoma before and after hepatic resection. Hepatogastroenterology. 1997 Mar-Apr;44(14):477-83
79 Kehrl JH, Roberts AB, Wakefield LM, et al. Transforming growth factor β is a important immunomodulatory protein for human B-lymphocytes. J Immunol.1986; 137:3855.
80 Shimizu S, Nishikawa Y, K uroda K, et al. Involvement of transforming growth factor β 1 in autocrine enhancement of gelatinase secretion by murine metastatic colon carcimoma cells. Cancer Res, 1996, 56:3366-3370
81 Bedossa P, Peltier E, Terris B, et al. Transforming growth factor betal (TGF-β_1) and TGF-β Receptor in normal, cirrhotic, and neoplastic human livers. Hepatology, 1995, 21 (3):760-766.
82 Matsumoto E, Nakatusukasa H, Nouso K, Kobayashi Y, Nakamura S, Suzuki M et al. Increased levels of tissue inhibitor of metalloproteinase-1 in human hepatocellular carcinoma. Liver int. 2004 Aug; 24(4):379-83
83 Miyoshi A, Kitajima Y, Sumi K, Soto K, Hagiwara A, Koga Y, Miyazaki K. Snail and SIP1 increase cancer invasion by upregulating MMP family in
84 ZHen HN, ZHang X, ZHang ZW, Bu XY, H. PZ, W ang BY. Expression of protooncogene c-erbB-1 and c-erbB-2 in human gliomas. Di-si Junyi Daxue Xuebao (J Fourth MiIMed U niv), 1999, 20 (2) 106-109
85 Hisaka T, Yano H haramaki M, et al. Expressions of epidernmal growth factor family and its receptor in hepatocellular carcimoma cell lines: relationship to cell proliferation [J]. Int J Oncol, 1999, 14:453
86 Arii S, Mori A, Uchida S, et al. Implication of vascular endothelial growth factor in the development and metastasis of human cancers.
87 Ueki T, fujimoto J, et al. Expression of hepatocyte growth factor and its receptor c-met proto-oncogene in hepatocellular carcinomal. [J]. Hepatology, 1997, 25(4):862-868.