HIF-1α对人肝癌细胞VEGF表达的影响
摘要
目的:缺氧是实质性肿瘤微环境的基本特征之一,同时肿瘤细胞的缺氧也是肿瘤发生恶性转化甚至转移的启动因子。缺氧诱导因子-l (hypoxia inducible factor-1, HIF-1 )是缺氧条件下广泛存在于哺乳动物和人体内的一种转录因子,由HIF-1α和HIF-1β两个亚单位组成,是调节细胞内氧代谢的关键因子之一,其中HIF-1α是主要的氧调节亚基。HIF-1作为一种核转录因子,在哺乳动物的生长发育、生理和病理反应过程中起重要作用。研究证实HIF-1α在肝癌等多种恶性肿瘤中普遍高表达,且与肿瘤的侵袭转移及预后等生物学特性有关。VEGF是HIF-1α的一个主要靶基因,通过调节VEGF的表达、增加肝细胞癌新生血管的生成,从而促进肝细胞癌的侵袭转移。HIF-1活性的调节,可能是治疗很多疾病的切入点。目前有关HIF-1α和VEGF关系的深入研究及以HIF-1α为治疗靶点的研究较少。本研究分别应用HIF-1α抑制剂YC-1[即3-(5'-羟甲基-2'-呋喃基)-1苯甲基引唑]及HIF-1α反义寡核苷酸作用于肝癌细胞株,检测药物干预前后肝癌细胞株SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF mRNA和蛋白表达的变化,明确HIF-1α对VEGF表达的影响,以及以HIF-1α为治疗靶点的可行性。
方法:肝癌细胞株SMMC-7721细胞、BEL-7402细胞分对照组和试验组。对照组细胞常规培养,试验组:分别用HIF-1α抑制剂YC-1、HIF-1α反义寡核苷酸作用于人肝癌细胞,培养24小时后。采用逆转录-聚合酶链式反应(RT-PCR)技术,测定干预前后肝癌细胞株SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF mRNA表达的变化,以β-actin作为内参照标准,通过凝胶扫描仪对扩增产物进行DNA电泳条带的光密度值分析,将HIF-1α、VEGF分别与β-actin比值作为HIF-1α、VEGF mRNA表达水平的参数,对HIF-1α、VEGF mRNA产物相对定量;以Western blot方法检测SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF蛋白表达的变化,同样以β-actin作为内参照标准,对HIF-1α、VEGF蛋白进行相对定量分析。所有数据均以x±s表示,采用SPSS13.0统计分析软件进行单因素方差分析(ANOVA)和t检验。P<0.05为有显著差异。
结果:1在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,对照组SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF mRNA的表达显著高于实验组(P<0.05)。
2在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,对照组SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF蛋白的表达显著高于实验组(P<0.05)。。
3在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞HIF-1αmRNA的表达量逐渐减少,组间比较差异具有显著性(P<0.05);随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞VEGF的表达产物VEGF_(121)(541bp)和VEGF_(165) (408bp)的表达量亦呈减少趋势,组间比较差异具有显著性(P<0.05),其中VEGF_(165)的表达要明显高于VEGF_(121)。
4在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞HIF-1α蛋白的表达量逐渐减少,组间比较差异具有显著性(P<0.05);随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞VEGF蛋白的表达亦呈减少趋势,组间比较差异具有显著性(P<0.05)。
结论:1 HIF-1α、VEGF在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中高表达。说明HIF-1α和VEGF均与肝细胞癌的发生、发展有关。
2 YC-1可以在基因和蛋白水平上抑制HIF-1α的表达,并下调VEGF的表达。同样HIF-1α反义寡核苷酸对肝癌细胞HIF-1α表达亦有抑制作用,同时VEGF的表达下降。说明HIF-1α可以促进肝癌细胞VEGF的表达。
3在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞HIF-1α和VEGF的表达下降,说明针对HIF-1α为靶点的治疗是可行的。
Objective:Hypoxia is a common characteristic of locally advanced solid tumors that has been associated with diminished therapeutic response, and more recently, with malignant progression, that is, an increasing probability of recurrence, loco regional spread, and distant metastasis. The hypoxia inducible factor 1(HIF-1) is a heterodimer transcription factor that is an important regulator of the growing tumor’s response to hypoxia in mammal or human body. It consists of HIF-1αand HIF-1βsubunits, HIF-1 activity in tumors depends on the availability of the HIF-1αsubunit. As nuclear transcription factor, HIF-1 are important in mammal of regulating the growing development, physiology and pathology. The study indicated: HIF-1α, which has high expression in hepatocellular carcinoma and in other malignant tumors, is associated with the tumor aggressiveness, migration and mortality. VEGF, as a target gene of HIF-1α, stimulating angiogenesis, promoting the aggressiveness and migration in hepatocellular carcinoma, is regulated by HIF-1α. The activity of HIF-1 may be the key to cure the malignant tumors. At the present time, there was little deeply study about relation of HIF-1αand VEGF in clinical research. In this research, we explored whether VEGF can be up-regulated by HIF-1αin hepatocellular carcinoma. The cell lines of SMMC-7721 and BEL-7402 were treated with YC-1 [3-(5'- hydroxymethyl-2'-furyl)-1-benzylindazole] and ASODN of HIF-1αrespectively. By detecting the expression of HIF-1α、VEGFmRNA and protein in SMMC- 7721 cell and BEL-7402 cell culturing in normal and treated conditions by RT-PCR and Western blot, we investigate the relation of HIF-1αand VEGF, and the potential of HIF-1αas a valid therapeutic target.
Methods: SMMC-7721 cells and BEL-7402 cells were divided as control group and experiment group. The control group were cultured with normal condition, the experiment group were treated by YC-1 or ASODN of HIF-1αrespectively with various concentration. All of the cells were collected on time (after cultured 24 hours). The expression of HIF-1α、VEGF mRNA levels was evaluated by RT-PCR. The HIF-1α、VEGF protein levels in cytoplasm was evaluated by western blot. The OD value of electrophoresis strip of DNA product or that of HIF-1α、VEGF protein was analyzed by gel scanner. Beta-actin was used as an internal standard. The relative expression level of HIF-1α、VEGF was represented with the ratio between produces of HIF-1α、VEGF and that of beta-actin. Data were analyzed with ANOVA and T test using SPSS13.0 statistical software. A level of P<0.05 was considered statistically significant.
Results: 1 In SMMC-7721 cells and BEL-7402 cells, the expression of HIF-1αand VEGF mRNA in experiment group were obviously down-regulated compared with control group, the differences of the expression of HIF-1αand VEGF mRNA were statistically significant (P<0.05).
2 In SMMC-7721 cells and BEL-7402 cells, the expression of HIF-1αand VEGF protein in experiment group were obviously down-regulated compared with control group, the differences of the expression of HIF-1αand VEGF protein were statistically significant (P<0.05).
3 In SMMC-7721 cells and BEL-7402 cells, following the increasing of the concentration of YC-1 or ASODN of HIF-1α, the expression of HIF-1αmRNA were decreased gradually, and the differences of the expression of HIF-1αmRNA were statistically significant within the experiment group(P<0.05). The expression of HIF-1αmRNA showed a dose-dependent in cells cultured with YC-1 or ASODN of HIF-1α. The expression of VEGF mRNA (including VEGF121 and VEGF165) were decreased gradually in experiment group, the differences of the expression of VEGF mRNA were statistically significant (P<0.05). VEGF165 mRNA has high expression compared with VEGF121 mRNA.
4 In experiment group of SMMC-7721 cells and BEL-7402 cells, following the increasing of the concentration of YC-1 or ASODN of HIF-1α, the expression of HIF-1αprotein showed decrease, it was statistically significant (P<0.05), and it showed a dose-dependent in cells cultured with YC-1 or ASODN of HIF-1α. The expression of VEGF protein were obviously down-regulated in cells cultured with YC-1 or ASODN of HIF-1α, it was statistically significant, too (P<0.05).
Conclutions: 1 The high expression of HIF-1αand VEGF in SMMC-7721 cells and BEL-7402 cells indicated that HIF-1αand VEGF were associated with the occurrence and development of HCC.
2 YC-1 inhibits the expression of HIF-1αat gene level and protein level. The expression of VEGF was down-regulated at gene level and protein level. ASODN of HIF-1αcould inhibited the expression of HIF-1αand VEGF at gene level and protein level, too. The result indicated that HIF-1αcan promote the expression of VEGF at gene level and protein level, in other word, that the expression of HIF-1αcan up-regulated the expression of VEGF in hepatocellular carcinoma.
3 Following the increasing of the concentration of YC-1 or ASODN of HIF-1αin experiment group in SMMC-7721 cells and BEL-7402 cells, the expression of HIF-1αand VEGF were decreased gradually. The result indicated that it has high potential to treatment tumor of HIF-1αas a valid therapeutic target.
方法:肝癌细胞株SMMC-7721细胞、BEL-7402细胞分对照组和试验组。对照组细胞常规培养,试验组:分别用HIF-1α抑制剂YC-1、HIF-1α反义寡核苷酸作用于人肝癌细胞,培养24小时后。采用逆转录-聚合酶链式反应(RT-PCR)技术,测定干预前后肝癌细胞株SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF mRNA表达的变化,以β-actin作为内参照标准,通过凝胶扫描仪对扩增产物进行DNA电泳条带的光密度值分析,将HIF-1α、VEGF分别与β-actin比值作为HIF-1α、VEGF mRNA表达水平的参数,对HIF-1α、VEGF mRNA产物相对定量;以Western blot方法检测SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF蛋白表达的变化,同样以β-actin作为内参照标准,对HIF-1α、VEGF蛋白进行相对定量分析。所有数据均以x±s表示,采用SPSS13.0统计分析软件进行单因素方差分析(ANOVA)和t检验。P<0.05为有显著差异。
结果:1在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,对照组SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF mRNA的表达显著高于实验组(P<0.05)。
2在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,对照组SMMC-7721细胞、BEL-7402细胞HIF-1α、VEGF蛋白的表达显著高于实验组(P<0.05)。。
3在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞HIF-1αmRNA的表达量逐渐减少,组间比较差异具有显著性(P<0.05);随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞VEGF的表达产物VEGF_(121)(541bp)和VEGF_(165) (408bp)的表达量亦呈减少趋势,组间比较差异具有显著性(P<0.05),其中VEGF_(165)的表达要明显高于VEGF_(121)。
4在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞HIF-1α蛋白的表达量逐渐减少,组间比较差异具有显著性(P<0.05);随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞VEGF蛋白的表达亦呈减少趋势,组间比较差异具有显著性(P<0.05)。
结论:1 HIF-1α、VEGF在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中高表达。说明HIF-1α和VEGF均与肝细胞癌的发生、发展有关。
2 YC-1可以在基因和蛋白水平上抑制HIF-1α的表达,并下调VEGF的表达。同样HIF-1α反义寡核苷酸对肝癌细胞HIF-1α表达亦有抑制作用,同时VEGF的表达下降。说明HIF-1α可以促进肝癌细胞VEGF的表达。
3在肝癌细胞株SMMC-7721细胞、BEL-7402细胞中,随着YC-1或HIF-1α反义寡核苷酸浓度的增加实验组细胞HIF-1α和VEGF的表达下降,说明针对HIF-1α为靶点的治疗是可行的。
Objective:Hypoxia is a common characteristic of locally advanced solid tumors that has been associated with diminished therapeutic response, and more recently, with malignant progression, that is, an increasing probability of recurrence, loco regional spread, and distant metastasis. The hypoxia inducible factor 1(HIF-1) is a heterodimer transcription factor that is an important regulator of the growing tumor’s response to hypoxia in mammal or human body. It consists of HIF-1αand HIF-1βsubunits, HIF-1 activity in tumors depends on the availability of the HIF-1αsubunit. As nuclear transcription factor, HIF-1 are important in mammal of regulating the growing development, physiology and pathology. The study indicated: HIF-1α, which has high expression in hepatocellular carcinoma and in other malignant tumors, is associated with the tumor aggressiveness, migration and mortality. VEGF, as a target gene of HIF-1α, stimulating angiogenesis, promoting the aggressiveness and migration in hepatocellular carcinoma, is regulated by HIF-1α. The activity of HIF-1 may be the key to cure the malignant tumors. At the present time, there was little deeply study about relation of HIF-1αand VEGF in clinical research. In this research, we explored whether VEGF can be up-regulated by HIF-1αin hepatocellular carcinoma. The cell lines of SMMC-7721 and BEL-7402 were treated with YC-1 [3-(5'- hydroxymethyl-2'-furyl)-1-benzylindazole] and ASODN of HIF-1αrespectively. By detecting the expression of HIF-1α、VEGFmRNA and protein in SMMC- 7721 cell and BEL-7402 cell culturing in normal and treated conditions by RT-PCR and Western blot, we investigate the relation of HIF-1αand VEGF, and the potential of HIF-1αas a valid therapeutic target.
Methods: SMMC-7721 cells and BEL-7402 cells were divided as control group and experiment group. The control group were cultured with normal condition, the experiment group were treated by YC-1 or ASODN of HIF-1αrespectively with various concentration. All of the cells were collected on time (after cultured 24 hours). The expression of HIF-1α、VEGF mRNA levels was evaluated by RT-PCR. The HIF-1α、VEGF protein levels in cytoplasm was evaluated by western blot. The OD value of electrophoresis strip of DNA product or that of HIF-1α、VEGF protein was analyzed by gel scanner. Beta-actin was used as an internal standard. The relative expression level of HIF-1α、VEGF was represented with the ratio between produces of HIF-1α、VEGF and that of beta-actin. Data were analyzed with ANOVA and T test using SPSS13.0 statistical software. A level of P<0.05 was considered statistically significant.
Results: 1 In SMMC-7721 cells and BEL-7402 cells, the expression of HIF-1αand VEGF mRNA in experiment group were obviously down-regulated compared with control group, the differences of the expression of HIF-1αand VEGF mRNA were statistically significant (P<0.05).
2 In SMMC-7721 cells and BEL-7402 cells, the expression of HIF-1αand VEGF protein in experiment group were obviously down-regulated compared with control group, the differences of the expression of HIF-1αand VEGF protein were statistically significant (P<0.05).
3 In SMMC-7721 cells and BEL-7402 cells, following the increasing of the concentration of YC-1 or ASODN of HIF-1α, the expression of HIF-1αmRNA were decreased gradually, and the differences of the expression of HIF-1αmRNA were statistically significant within the experiment group(P<0.05). The expression of HIF-1αmRNA showed a dose-dependent in cells cultured with YC-1 or ASODN of HIF-1α. The expression of VEGF mRNA (including VEGF121 and VEGF165) were decreased gradually in experiment group, the differences of the expression of VEGF mRNA were statistically significant (P<0.05). VEGF165 mRNA has high expression compared with VEGF121 mRNA.
4 In experiment group of SMMC-7721 cells and BEL-7402 cells, following the increasing of the concentration of YC-1 or ASODN of HIF-1α, the expression of HIF-1αprotein showed decrease, it was statistically significant (P<0.05), and it showed a dose-dependent in cells cultured with YC-1 or ASODN of HIF-1α. The expression of VEGF protein were obviously down-regulated in cells cultured with YC-1 or ASODN of HIF-1α, it was statistically significant, too (P<0.05).
Conclutions: 1 The high expression of HIF-1αand VEGF in SMMC-7721 cells and BEL-7402 cells indicated that HIF-1αand VEGF were associated with the occurrence and development of HCC.
2 YC-1 inhibits the expression of HIF-1αat gene level and protein level. The expression of VEGF was down-regulated at gene level and protein level. ASODN of HIF-1αcould inhibited the expression of HIF-1αand VEGF at gene level and protein level, too. The result indicated that HIF-1αcan promote the expression of VEGF at gene level and protein level, in other word, that the expression of HIF-1αcan up-regulated the expression of VEGF in hepatocellular carcinoma.
3 Following the increasing of the concentration of YC-1 or ASODN of HIF-1αin experiment group in SMMC-7721 cells and BEL-7402 cells, the expression of HIF-1αand VEGF were decreased gradually. The result indicated that it has high potential to treatment tumor of HIF-1αas a valid therapeutic target.
引文
1 黄耿文,杨连粤. 缺氧致肿瘤恶性转化的分子机制,世界华人消化杂志,2001 , 9 : 1300-1304
2 Sanchez Elsner T, Botella L M, Velasco B et al . Synergistic cooperation between hypoxia and transforming growth factor-β pathways on human vascular endothelial growth factor gene expression .J Biol Chem, 2001, 276: 38527-38535
3 Lee JW, Bae SH, Jeong JW, et a1. Hypoxia-induciblefactor (HIF-1)alpha:its protein stability and biological functions [J]. Exp Mol Med, 2004, 36(1):1- 12
4 王燕,汪睿等.缺氧诱导因子-1 与肿瘤治疗研究进展. Medical Recapitulate 2004, 10(1): 5-9
5 Yang QC, Zeng BF, Shi ZM Inhibition of hypoxia-induced angiogenesis by trichostatin A via sup-pression of HIF-lα activity in human osteo sarcoma. J Exp Clin Cancer Res. 2006, 25(4): 593-599
6 Na X, Wu G Ryan CK, et al. Overproduction of vascular endothelial growth factor related to von Hippel- lindautumor suppressor gene mutations and hypoxia- inducible factor-1 alpha expression in renal cell carcinomas .J Urol, 2003, 170(2 Pt 1): 588-592
7 Lidgren A, Hedberg Y, Grankvist K ,et al. The expression of hypoxia-inducible factor 1alpha is a favorable independent prognostic factor in renal cell carcinoma. Clin Cancer Res,2005, 11(3):1129-1135
8 Mizukami Y Li J, Zhang X, et al, Hypoxia-inducible factor-1 independent regulation of vascular endothelial growth factor by hypoxia in colon cancer. Cancer Res, 2004,64(5): 1765-1772
9 Bos R, van der Groep P, Greijer AE, et al. Levels of hypoxia-inducible factor 1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. Cancer, 2003, 97(6): 1573-1581
10 Buchler P, Reber HA, Buchler M, et al. Hypoxia-inducible factor 1 regulates vascular endothelial growth factor expression in human pancreatic cancer, Pancreas, 2003, 26(1): 56-64
11 Sivridis E, Giatromanolaki A, Gatter KC, et al. Association of hypoxia-inducible factor 1alpha and 2alpha with activated angiogenic pathways and prognosis in patients with endometrial carcinoma. Cancer, 2002, 95(5): 1055-1063
12 Koukourakis MI, Giatromanolaki A, Sivridis E, et al. Hypoxia-inducible factor (HIF1αand HIF2α), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer. Int J Radiat Oncol Biol Phys, 2002, 53(5): 1192-1202
13 Nakayama K, Kanzaki A, Hata K, et al. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) gene expression in human ovarian carcinoma. Cancer Lett, 2002, 176(2): 215-223
14 Huang GW, Yang LY, Lu WQ. Expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor in hepatocellular carcinoma: Impact on neovascul- arization and survival. World J Gastroenterol , 2005, 11(11): 1705-1708
15 Matsuyama T, Nakanishi K, Hayashi T, et al. Expression of hypoxia-inducible factor-1 alpha in esophageal squamous cell carcinoma. Cancer Sci, 2005, 96(3): 176-182
16 Lee CH, Lee MK, Kang CD, et al. Differential expression of hypoxia-inducible factor 1 alpha and tumor cell proliferation between squamous cell carcinomas and adenocarcinomas among operable non small cell lung carcinomas[J]. J Korean Med Sei, 2003, 18(2): 196-203
17 丁磊, 陈孝平, 王海平. 缺氧诱导因子-1α 蛋白在肝癌组织中的表达及临床意义. 中华肝胆病杂志, 2004, 12(11): 656-659
18 江从庆, 刘志苏, 钱群, 等. 大肠腺瘤和腺癌组织中缺氧诱导因子-1α 的表达及其与 VEGF、微血管密度的关系.癌症, 2003, 22(11): 1170-1174
19 裘正军, 蒋奕, 胡文慧, 等. HIF-1α、VEGF 在胰腺癌组织中的表达及其与血管新生的关系, 肿瘤, 2005, 25(2): 155-157
20 Sivridis E, Giatromanolaki A, Gatter K C, et al. Association of hypoxia-inducible factors 1 alpha and 2 alpha with activated angiogenic pathways and prognosis in patients with endometrial carcinoma [J]. Cancer, 2002, 95(5):1055
21 Liu LX, Lu H. Stabilization of vascular endothelial growthfactor mRNA by hypoxia-inducible factor 1. Biochem Biophys Res Commun, 2002, 291(4): 908-14
22 Buchler P, Reber HA, Buchler MW, et al. Antiangiogenic activity of genistein in pancreatic carcinoma cells is mediated by the inhibition of hypoxia-inducible factor-1 and the down-regu-lation of VEGF gene expression [J]. Cancer, 2004, 100(1):201-210
23 丁磊, 陈孝平, 王海平. 封闭缺氧诱导因子作用位点对人肝癌细胞生长抑制的研究. 华中科技大学学报(医学版), 2005,34(6): 751-753
24 Sun X, Kanwar JR , Leung E, et al .Gene transfer of antisense hypoxia inducible factor-1 alpha enhances the therapeutic efficacy of cancer immunotherapy . Gene Ther , 2001, 8 (8) : 638-645
25 顾永平, 谢芳, 何杨, 等. 胰腺癌细胞在低氧状态中HIF-1、VEGF 蛋白的表达及可溶性 VEGF 水平变化的生物学意义. 中国现代医药杂志, 2006, 11(8): 1-3
26 Yeo EJ, Chun YS, Park JW. New anticancer strategies targeting HIF-1[J]. Biochem Pharmacol, 2004, 68(6): 1061- 1069
27 Welsh S, Williams R, Kirkpatrick L, et al. Antitumor activity and pharmacodynamic properties of PX-478 , an inhibitor of hypoxia-inducible factor-1alpha . Mol Cancer Ther , 2004 , 3 ( 3 ) : 233-244
28 Yeo EJ, Chun YS, Cho YS, et al. YC-1: a potential anticancer drug targeting hypoxia-inducible factor 1. J Natl Cancer Inst2003, 95: 516-525
29 Chen CJ, Hsu MH, Huang LJ, et al. Anticancer mechanisms of YC-1 in human lung cancerc ell line, NCI-H226. Bioch- emical pharmacology, 2008, 75(2): 360-368
30 Wang SW, Pan SL, Guh JH, Chen HL, Huang DM,Chang YL, et al. YC-1 [3-(5'-hydroxymethyl-2'-furyl) -1- benzli- ndazole] exhibits a novel antiproliferative effect and arrests the cell cycle in G0-G1 in human hepatocellular carcinoma cells. J Pharmacol Exp Ther 2005; 312: 917-25
31 Yeo EJ, Ryu JH, Chun YS, Cho YS, Jang IJ, Cho H, et al. YC-1 induces S cell cycle arrest and apoptosis by activating checkpoint kinases. Cancer Res 2006; 66: 6345-52
32 Pan SL, Guh JH, Peng CY, Wang SW, Chang YL, Cheng FC, et al. YC-1 [3-(5'-hydroxymethyl-2'-furyl) -1- benzli- ndazole] inhibits endothelial cell functions induced by angiogenic factors in vitro and angiogenesis in vivo models. J Pharmacol Exp Ther 2005; 314: 35-42
33 Pan SL, Guh JH, Peng CY, Chang YL, Cheng FC, Chang JH, et al. A potential role of YC-1 on the inhibition of cytokine release in peripheral blood mononuclear leukocytes and endotoxemic mouse models. Thromb Haemost 2005; 93: 940-8
34 Liu YN, Pan SL, Peng CY, Guh JH, Huang DM, Chang YL, et al. YC-1 [3-(5'-hydroxymethyl-2'-furyl) -1- benzli-ndazole] inhibits neointima formation in balloon-injured rat carotid through suppression of expressions and activities of matrixmetal-oproteinases 2 and 9. J Pharmacol Exp Ther 2006; 316:35-41
35 Huang YT, Pan SL, Guh JH, Chang YL, Lee FY, Kuo SC, et al. YC-1 suppresses constitutive nuclear factor- kappa B activation and induces apoptosis in human prostate cancer cells. Mol Cancer Ther 2005; 4: 1628-35
1 Lee JW, Bae SH, Jeong JW, et a1. Hypoxia-inducible factor(HIF-1) alpha: its protein stability and biological functions[J]. Exp Mol Med, 2004,36(1):1-12
2 Huang LE, Gu J, Schau M, et al. Regulation of hypoxia inducible factor-1 alpha is mediated by an O2 dependent degradation domain via the ubiquitin-proteasome pathway [J]. Proc Natl Acad Sci USA, 1998,95(14):7987~7992
3 Semenza GL, Wang GL.A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation. Mol Cell Biol, 1992, 12(12):5447-54
4 Cassmann M , Chilov D , Wenger RH . Regulation of the hypoxia-inducible factor -1 alpha . ARNT is not necessary for hypoxic induction of HIF-1 alpha in the nucleus [J]. Adv Exp Med Biol , 2000, 475 : 87-99
5 Maxwell PH, Dachs GU, Gleadle JM, et al. Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogene-sis and tumor growth. Proc Natl Acad Sci USA, 1997, 94 (15): 8104-9
6 Goda N, Dozier SJ, Johnson RS. HIF-1 in Cell Cycle Regulation, Apoptosis, and Tumor Progression. Antioxid Redox Signal, 2003, 5: 467-473
7 Lorenz Poellinger, Randall S J. HIF-1 and hypoxic response:the plot thickens[J]. Genetics & Development, 2004, 14:81-85
8 Knowles HJ, Raval RR, Harris AL, et al. Effect of ascorbate on the activity of hypoxia-inducible factor in cancer cells. Cancer Res, 2003, 63: 1764-1768
9 Berra E, Benizri E, Ginouves A, et al. HIF prolyl-hydroxylase 2 is the key oxygen sensor setting low steady-state levels of HIF-1alpha in normoxia[J]. EMBO J, 2003,22:4082-4090
10 Ang S O, Chen H, Hirota K, et al. Disruption of oxygen homeostasis underlies congenital Chuvash polycythemia[J]. Nat Genet, 2002,32:614-621
11 Pugh CW, Ratcliffe PJ. Regulation of angiogenesis by hypoxia: role of the HIF system. Nat Med, 2003, 9:677-684.
12 Mazure NM, Brahimi-Horn MC, Pouyssegur J. Protein kinases and the hypoxia-inducible factor-1, two switches in angiogenesis. Curr Pharm Des, 2003, 9: 531-541
13 Hirota K. Hypoxia-inducible factor 1, a master transcription factor of cellular hypoxic gene expression. J Anesth, 2002, 16:150-159
14 Hirota K. Hypoxia-inducible factor 1, a master transcription factor of cellular hypoxic gene expression. J Anesth, 2002, 16:150-159
15 Liu Y, Cox SR, Morita T, et al. Hypoxia regulates vascular endothelial growth factor gene expression in endothelial cells: identification of a 5 enhancer [J]. Circ Res, 1995, 77 (3):638-643
16 Liu L X, L u H, L uo Y, et al. Stabilization of vascular endothelial growth factor mRNA by hypoxia-inducible factor 1. Biochem Biophys Res Commun, 2002, 291: 908-914
17 Stein I, Itin A, Einat P, et al. Translation of vascular endothelial growth factor mRNA by internal ribosome entry: implications for translation under hypoxia. MolCell Biol, 1998, 18: 3112-3119
18 鲍永霞,吕福祯,马迎军,等。低氧对小鼠低氧诱导因子-1α和血管内皮生长因子表达的影响。中国实用内科杂志,2006,26(14):1066-1068
19 Gerber HP, Condorelli F, Park J, et al. Differential transcriptional regulation of the two vascular endothelial growth factor receptor genes. Flt-1, but not Flk-1/KDR, is up-regulated by hypoxia. J Biol Chem, 1997,272: 23659-23667
20 Du JR, Jiang Y, Zhang YM, et al. VEGF and microvasgular density in esophageal and gastric carcinoimas[J]. World J Gastroenterol, 2003,9:1 604~1 606
21 Yang QC, Zeng BF, Shi ZM Inhibition of hypoxia-induced angiogenesis by tirchostatin A via suppression of HIF-la activity inhuman osteosarcoma.J Exp Clin Cancer Res 2006; 25(4): 593-599
22 Jiang DH, Guo DY, Li HZ. Inhibited proliferation and expression of human nasopharyngeal carcinoma line induced by celecoxib in vitro Zhonghua Er Bi Yan Hou TonJing Wai Ke Za Zhi. 2006,41(11):809-812
23 Mohamed KM, Le A, Duong H, Correlation between VEGF and HIF-lalpha expression in human oral squamous cell carcinoma. Exp Mol Pathol. 2004,76(2):143-152
24 Zhao L, Han B, Liu J, et al, Expression and significance of vascular endothelial growth factor in human esophageal cardinoma[j]. China journal of Modern Medicine, 2004, 14(2):59-61, 66. Chinese
25 Buchler P, Reber HA, Buchler M, et al. Hypoxia-inducible factor 1 regulates vascular endothelial growth factor expression in human pancreatic cancer, Pancreas, 2003, 26(1): 56-64
26 Ito A, Koshikawa N, Mochizuki S. Hypoxia-inducible factor-1 mediates the expression of DNA polymerase iota in human tumor cells. Biochem Biophys Res Commun. 2006,351(1): 306-311
27 Yasuda S, Arii S, Mori A, Isobe N, et al. Hexokinse Ⅱ and VEGF expression in liver tumors: correlation with hypoxia-inducible factor 1 alpha and its significance[J] J Hepatol. 2004,40(1):117-123
28 王燕,汪睿等.缺氧诱导因子-1 与肿瘤治疗研究进展. Medical Recapitulate 2004, 10(1): 5-9
29 Yang QC, Zeng BF, Shi ZM Inhibition of hypoxia-induced angiogenesis by trichostatin A via sup-pression of HIF-lα activity in human osteo sarcoma. J Exp Clin Cancer Res. 2006, 25(4):593-599
30 丁磊,陈孝平,王海平。肝癌及癌旁组织中 HIF-1α基因 蛋白的表达及其临床意义。肝胆外科杂志, 2004, 12(1):32-35
31 Semenza GL. O2-regulated gene expression: transcript- tional control of cardiorespiratory physiology by HIF-1[J]. J Appl Physiol,2004,96(3):1173-1177
32 Na X, Wu G Ryan CK, et al. Overproduction of vascular endothelial growth factor related to von Hippel- lindautumor suppressor gene mutations and hypoxia- inducible factor-1 alpha expression in renal cell carcinomas .J Urol, 2003, 170(2 Pt 1): 588-592
33 Lidgren A, Hedberg Y, Grankvist K ,et al. The expression of hypoxia-inducible factor 1alpha is a favorable independent prognostic factor in renal cell carcinoma. Clin Cancer Res, 2005, 11(3):1129-1135
34 Mizukami Y Li J, Zhang X, et al, Hypoxia-inducible factor-1 independent regulation of vascular endothelial growth factor by hypoxia in colon cancer. Cancer Res, 2004,64(5): 1765-1772
35 Bos R, van der Groep P, Greijer AE, et al. Levels of hypoxia-inducible factor 1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. Cancer, 2003, 97(6): 1573-1581
36 Sivridis E, Giatromanolaki A, Gatter KC, et al. Association of hypoxia-inducible factor 1alpha and 2alpha with activated angiogenic pathways and prognosis in patientswith endometrial carcinoma. Cancer, 2002, 95(5): 1055-1063
37 Koukourakis MI, Giatromanolaki A, Sivridis E, et al. Hypoxia-inducible factor (HIF1αand HIF2α), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer. Int J Radiat Oncol Biol Phys, 2002, 53(5): 1192-1202
38 Nakayama K, Kanzaki A, Hata K, et al. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) gene expression in human ovarian carcinoma. Cancer Lett, 2002, 176(2): 215-223
39 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(11): 1705-1708
40 Lee CH, Lee MK, Kang CD, et al. Differential expression of hypoxia-inducible factor 1 alpha and tumor cell proliferation between squamous cell carcinomas and adenocarcinomas among operable non small cell lung carcinomas[J]. J Korean Med Sei, 2003, 18(2): 196-203
41 王劲红,李福琴,艾丽敏等.宫颈癌中缺氧诱导因子-1α 的表 达 和 血 管 生 成 的 关 系 . 中 国 肿 瘤 与 临 床 , 2004, 24:1426-1427
42 Yu EZ, Li YY, Liu XH, et al, Antiapoptotic action of hypoxia-inducible factor-1 alpha in human endothelial cells [J]. Lab invest 2004, 84950:553-516
43 王晗,韩忠朝. 缺氧诱导因子-1 与细胞凋亡[J]. 国外医学生理、病理科学与临床分册,2005,1(15):52-55
44 Sasabe E, Tatemoto Y, Li D, et al. Mechanism of HIF-l alpha dependen Suppression of hypoxia-induced apoptosis in squamous cell carcinoma cells[J]. Cancer Sci, 2005, 96(7):394-402
45 Kim JY, Ahn HJ, Ryu JH, et al. BH3-only protein noxa is a mediator of hypoxic cell death induced by hypoxia- inducible factor 1 alpha [J]. J Exp Med, 2004, 199(1): 113-124
46 Lee MI, Kim JY, Suk K, et al. Identification of the hypoxia-inducible factor 1 alpha-responsive HGTD-P gene as a mediator in mitochondrial apoptotic pathway [J]. Mol Cell Biol, 2004, 24(9):3918-3127
47 Semenza GL. Targeting HIF-1 for cancer therapy[J]. Nat Rev Cancer, 2003, 3(10): 721-732
48 Xia S,Yu SY, Yuan XL,et al. Effects of hypoxia on expression of P-glycoprotein and multidrug resistance Protein in human lung adenocarcinoma A549 ecll line[J]. Chin Mde J,2004,84(8):663- 666
49 Wartenberg M, Ling FC,Muschen M,et al. Regulation of the multidrug resistance transporter P-glycoprotein in multicellular tumor spheroids by hypoxia-inducible factor 1(HIF-1)and reactive oxygen species[J]. FASEB J, 2003, 17(3): 503-505
50 Thorpe P. Antibody directed targeting of tumor vasculature.Proc Am Assoc Cancer Res, 1996, 37: 668
51 Mahendra G, KumarS, IsayevaT, et al. Antiangiogenic cancer gene therapy by adeno-associated virus 2-mediated stable expression of the soluble FMS-like tyrosine kinase-1 receptor. Cancer Gene Ther, 2004 Sep 10
52 Qian DZ, Wang X, Kachlmp SK, et al. The Histone Deacetylase Inhibitor NVP_LAQ824 Inhibits Angiogenesis and Has a Greater Antitumor Effect in Combination with the Vascular Endothelial Growth Factor Receptor Tyro-sine Kinase Inhibitor PTK787/ ZK222584. CancerRes, 2004, 64(18):6626-6634
53 Huckle WR, Roche RI. Post-transcriptional control of expression of sFlt-1, an endogenous inhibitor of vascular endothelial growth factor. J Cell Biochem, 2004, 93(1):120-132
54 Welsh SJ, Williams R, Kirkpatrick L, et al. Antitumor activity and pharmacodynamic properties of PX-478, an inhibitor of hypoxia-inducible factor-1 alpha. Mol Cancer Ther. 2004, 3(3):233-244
55 Mackenzie ED, Selak MA, Tennant DA, Cell permeating {alpha)-ketoglutarate derivatives alleviate pseudo-hypoxia in SDH deficient cells. Mol Cell Biol. 2007, 17(32):41-50
56 Rapisarda A, Aalek J, Hollinggshead M, et al. Schedule dependent inhibition of hypoxia-inducible factor-l alpha protein accumulation, angiogenesis, and tumor growth by topotecan in U251-HRE glioblastoma xenogragts. CancerRes, 2004, 64(19):6845-6848
57 Stoeltzing O, McCasrty MF, Wey JS, et al. Role of hypoxia-inducible factor l alpha in gastric cancer cell growth, angiogenesis, and vessel maturation. J Natl Cancer I, 2004, 96(12): 946-956
58 Yeo EJ, Chun YS, Cho YS, et al. YC-1: a potential anticancer drug targeting hypoxia-inducible factor 1. J Natl Cancer Inst 2003; 95: 516-525
59 Asc G, Xu X, Chu C, et al. Prognosic significance of erythropoieth expression in human endometrial carcinoma. Cancer, 2004, 100(11): 2376-2386
60 董典宁、孙平、智绪亭, 等, 反义 HIF-1α 基因治疗人肝癌裸鼠移植瘤的实验研究. 中国普通外科杂志,2006, 15(7):520-524
61 丁磊,陈孝平,王海平。封闭缺氧诱导因子作用位点对人肝癌细胞生长抑制的研究。华中科技大学学报(医学版),2005, 34(6):751-753
62 Li L, Lin X, Staver M, Evaluating hypoxia-inducible factor-l alpha as a cancer therapeutic target via inducible RNA interference in vivo. Cancer Res. 2005, 65(16): 7249-7258
2 Sanchez Elsner T, Botella L M, Velasco B et al . Synergistic cooperation between hypoxia and transforming growth factor-β pathways on human vascular endothelial growth factor gene expression .J Biol Chem, 2001, 276: 38527-38535
3 Lee JW, Bae SH, Jeong JW, et a1. Hypoxia-induciblefactor (HIF-1)alpha:its protein stability and biological functions [J]. Exp Mol Med, 2004, 36(1):1- 12
4 王燕,汪睿等.缺氧诱导因子-1 与肿瘤治疗研究进展. Medical Recapitulate 2004, 10(1): 5-9
5 Yang QC, Zeng BF, Shi ZM Inhibition of hypoxia-induced angiogenesis by trichostatin A via sup-pression of HIF-lα activity in human osteo sarcoma. J Exp Clin Cancer Res. 2006, 25(4): 593-599
6 Na X, Wu G Ryan CK, et al. Overproduction of vascular endothelial growth factor related to von Hippel- lindautumor suppressor gene mutations and hypoxia- inducible factor-1 alpha expression in renal cell carcinomas .J Urol, 2003, 170(2 Pt 1): 588-592
7 Lidgren A, Hedberg Y, Grankvist K ,et al. The expression of hypoxia-inducible factor 1alpha is a favorable independent prognostic factor in renal cell carcinoma. Clin Cancer Res,2005, 11(3):1129-1135
8 Mizukami Y Li J, Zhang X, et al, Hypoxia-inducible factor-1 independent regulation of vascular endothelial growth factor by hypoxia in colon cancer. Cancer Res, 2004,64(5): 1765-1772
9 Bos R, van der Groep P, Greijer AE, et al. Levels of hypoxia-inducible factor 1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. Cancer, 2003, 97(6): 1573-1581
10 Buchler P, Reber HA, Buchler M, et al. Hypoxia-inducible factor 1 regulates vascular endothelial growth factor expression in human pancreatic cancer, Pancreas, 2003, 26(1): 56-64
11 Sivridis E, Giatromanolaki A, Gatter KC, et al. Association of hypoxia-inducible factor 1alpha and 2alpha with activated angiogenic pathways and prognosis in patients with endometrial carcinoma. Cancer, 2002, 95(5): 1055-1063
12 Koukourakis MI, Giatromanolaki A, Sivridis E, et al. Hypoxia-inducible factor (HIF1αand HIF2α), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer. Int J Radiat Oncol Biol Phys, 2002, 53(5): 1192-1202
13 Nakayama K, Kanzaki A, Hata K, et al. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) gene expression in human ovarian carcinoma. Cancer Lett, 2002, 176(2): 215-223
14 Huang GW, Yang LY, Lu WQ. Expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor in hepatocellular carcinoma: Impact on neovascul- arization and survival. World J Gastroenterol , 2005, 11(11): 1705-1708
15 Matsuyama T, Nakanishi K, Hayashi T, et al. Expression of hypoxia-inducible factor-1 alpha in esophageal squamous cell carcinoma. Cancer Sci, 2005, 96(3): 176-182
16 Lee CH, Lee MK, Kang CD, et al. Differential expression of hypoxia-inducible factor 1 alpha and tumor cell proliferation between squamous cell carcinomas and adenocarcinomas among operable non small cell lung carcinomas[J]. J Korean Med Sei, 2003, 18(2): 196-203
17 丁磊, 陈孝平, 王海平. 缺氧诱导因子-1α 蛋白在肝癌组织中的表达及临床意义. 中华肝胆病杂志, 2004, 12(11): 656-659
18 江从庆, 刘志苏, 钱群, 等. 大肠腺瘤和腺癌组织中缺氧诱导因子-1α 的表达及其与 VEGF、微血管密度的关系.癌症, 2003, 22(11): 1170-1174
19 裘正军, 蒋奕, 胡文慧, 等. HIF-1α、VEGF 在胰腺癌组织中的表达及其与血管新生的关系, 肿瘤, 2005, 25(2): 155-157
20 Sivridis E, Giatromanolaki A, Gatter K C, et al. Association of hypoxia-inducible factors 1 alpha and 2 alpha with activated angiogenic pathways and prognosis in patients with endometrial carcinoma [J]. Cancer, 2002, 95(5):1055
21 Liu LX, Lu H. Stabilization of vascular endothelial growthfactor mRNA by hypoxia-inducible factor 1. Biochem Biophys Res Commun, 2002, 291(4): 908-14
22 Buchler P, Reber HA, Buchler MW, et al. Antiangiogenic activity of genistein in pancreatic carcinoma cells is mediated by the inhibition of hypoxia-inducible factor-1 and the down-regu-lation of VEGF gene expression [J]. Cancer, 2004, 100(1):201-210
23 丁磊, 陈孝平, 王海平. 封闭缺氧诱导因子作用位点对人肝癌细胞生长抑制的研究. 华中科技大学学报(医学版), 2005,34(6): 751-753
24 Sun X, Kanwar JR , Leung E, et al .Gene transfer of antisense hypoxia inducible factor-1 alpha enhances the therapeutic efficacy of cancer immunotherapy . Gene Ther , 2001, 8 (8) : 638-645
25 顾永平, 谢芳, 何杨, 等. 胰腺癌细胞在低氧状态中HIF-1、VEGF 蛋白的表达及可溶性 VEGF 水平变化的生物学意义. 中国现代医药杂志, 2006, 11(8): 1-3
26 Yeo EJ, Chun YS, Park JW. New anticancer strategies targeting HIF-1[J]. Biochem Pharmacol, 2004, 68(6): 1061- 1069
27 Welsh S, Williams R, Kirkpatrick L, et al. Antitumor activity and pharmacodynamic properties of PX-478 , an inhibitor of hypoxia-inducible factor-1alpha . Mol Cancer Ther , 2004 , 3 ( 3 ) : 233-244
28 Yeo EJ, Chun YS, Cho YS, et al. YC-1: a potential anticancer drug targeting hypoxia-inducible factor 1. J Natl Cancer Inst2003, 95: 516-525
29 Chen CJ, Hsu MH, Huang LJ, et al. Anticancer mechanisms of YC-1 in human lung cancerc ell line, NCI-H226. Bioch- emical pharmacology, 2008, 75(2): 360-368
30 Wang SW, Pan SL, Guh JH, Chen HL, Huang DM,Chang YL, et al. YC-1 [3-(5'-hydroxymethyl-2'-furyl) -1- benzli- ndazole] exhibits a novel antiproliferative effect and arrests the cell cycle in G0-G1 in human hepatocellular carcinoma cells. J Pharmacol Exp Ther 2005; 312: 917-25
31 Yeo EJ, Ryu JH, Chun YS, Cho YS, Jang IJ, Cho H, et al. YC-1 induces S cell cycle arrest and apoptosis by activating checkpoint kinases. Cancer Res 2006; 66: 6345-52
32 Pan SL, Guh JH, Peng CY, Wang SW, Chang YL, Cheng FC, et al. YC-1 [3-(5'-hydroxymethyl-2'-furyl) -1- benzli- ndazole] inhibits endothelial cell functions induced by angiogenic factors in vitro and angiogenesis in vivo models. J Pharmacol Exp Ther 2005; 314: 35-42
33 Pan SL, Guh JH, Peng CY, Chang YL, Cheng FC, Chang JH, et al. A potential role of YC-1 on the inhibition of cytokine release in peripheral blood mononuclear leukocytes and endotoxemic mouse models. Thromb Haemost 2005; 93: 940-8
34 Liu YN, Pan SL, Peng CY, Guh JH, Huang DM, Chang YL, et al. YC-1 [3-(5'-hydroxymethyl-2'-furyl) -1- benzli-ndazole] inhibits neointima formation in balloon-injured rat carotid through suppression of expressions and activities of matrixmetal-oproteinases 2 and 9. J Pharmacol Exp Ther 2006; 316:35-41
35 Huang YT, Pan SL, Guh JH, Chang YL, Lee FY, Kuo SC, et al. YC-1 suppresses constitutive nuclear factor- kappa B activation and induces apoptosis in human prostate cancer cells. Mol Cancer Ther 2005; 4: 1628-35
1 Lee JW, Bae SH, Jeong JW, et a1. Hypoxia-inducible factor(HIF-1) alpha: its protein stability and biological functions[J]. Exp Mol Med, 2004,36(1):1-12
2 Huang LE, Gu J, Schau M, et al. Regulation of hypoxia inducible factor-1 alpha is mediated by an O2 dependent degradation domain via the ubiquitin-proteasome pathway [J]. Proc Natl Acad Sci USA, 1998,95(14):7987~7992
3 Semenza GL, Wang GL.A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation. Mol Cell Biol, 1992, 12(12):5447-54
4 Cassmann M , Chilov D , Wenger RH . Regulation of the hypoxia-inducible factor -1 alpha . ARNT is not necessary for hypoxic induction of HIF-1 alpha in the nucleus [J]. Adv Exp Med Biol , 2000, 475 : 87-99
5 Maxwell PH, Dachs GU, Gleadle JM, et al. Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogene-sis and tumor growth. Proc Natl Acad Sci USA, 1997, 94 (15): 8104-9
6 Goda N, Dozier SJ, Johnson RS. HIF-1 in Cell Cycle Regulation, Apoptosis, and Tumor Progression. Antioxid Redox Signal, 2003, 5: 467-473
7 Lorenz Poellinger, Randall S J. HIF-1 and hypoxic response:the plot thickens[J]. Genetics & Development, 2004, 14:81-85
8 Knowles HJ, Raval RR, Harris AL, et al. Effect of ascorbate on the activity of hypoxia-inducible factor in cancer cells. Cancer Res, 2003, 63: 1764-1768
9 Berra E, Benizri E, Ginouves A, et al. HIF prolyl-hydroxylase 2 is the key oxygen sensor setting low steady-state levels of HIF-1alpha in normoxia[J]. EMBO J, 2003,22:4082-4090
10 Ang S O, Chen H, Hirota K, et al. Disruption of oxygen homeostasis underlies congenital Chuvash polycythemia[J]. Nat Genet, 2002,32:614-621
11 Pugh CW, Ratcliffe PJ. Regulation of angiogenesis by hypoxia: role of the HIF system. Nat Med, 2003, 9:677-684.
12 Mazure NM, Brahimi-Horn MC, Pouyssegur J. Protein kinases and the hypoxia-inducible factor-1, two switches in angiogenesis. Curr Pharm Des, 2003, 9: 531-541
13 Hirota K. Hypoxia-inducible factor 1, a master transcription factor of cellular hypoxic gene expression. J Anesth, 2002, 16:150-159
14 Hirota K. Hypoxia-inducible factor 1, a master transcription factor of cellular hypoxic gene expression. J Anesth, 2002, 16:150-159
15 Liu Y, Cox SR, Morita T, et al. Hypoxia regulates vascular endothelial growth factor gene expression in endothelial cells: identification of a 5 enhancer [J]. Circ Res, 1995, 77 (3):638-643
16 Liu L X, L u H, L uo Y, et al. Stabilization of vascular endothelial growth factor mRNA by hypoxia-inducible factor 1. Biochem Biophys Res Commun, 2002, 291: 908-914
17 Stein I, Itin A, Einat P, et al. Translation of vascular endothelial growth factor mRNA by internal ribosome entry: implications for translation under hypoxia. MolCell Biol, 1998, 18: 3112-3119
18 鲍永霞,吕福祯,马迎军,等。低氧对小鼠低氧诱导因子-1α和血管内皮生长因子表达的影响。中国实用内科杂志,2006,26(14):1066-1068
19 Gerber HP, Condorelli F, Park J, et al. Differential transcriptional regulation of the two vascular endothelial growth factor receptor genes. Flt-1, but not Flk-1/KDR, is up-regulated by hypoxia. J Biol Chem, 1997,272: 23659-23667
20 Du JR, Jiang Y, Zhang YM, et al. VEGF and microvasgular density in esophageal and gastric carcinoimas[J]. World J Gastroenterol, 2003,9:1 604~1 606
21 Yang QC, Zeng BF, Shi ZM Inhibition of hypoxia-induced angiogenesis by tirchostatin A via suppression of HIF-la activity inhuman osteosarcoma.J Exp Clin Cancer Res 2006; 25(4): 593-599
22 Jiang DH, Guo DY, Li HZ. Inhibited proliferation and expression of human nasopharyngeal carcinoma line induced by celecoxib in vitro Zhonghua Er Bi Yan Hou TonJing Wai Ke Za Zhi. 2006,41(11):809-812
23 Mohamed KM, Le A, Duong H, Correlation between VEGF and HIF-lalpha expression in human oral squamous cell carcinoma. Exp Mol Pathol. 2004,76(2):143-152
24 Zhao L, Han B, Liu J, et al, Expression and significance of vascular endothelial growth factor in human esophageal cardinoma[j]. China journal of Modern Medicine, 2004, 14(2):59-61, 66. Chinese
25 Buchler P, Reber HA, Buchler M, et al. Hypoxia-inducible factor 1 regulates vascular endothelial growth factor expression in human pancreatic cancer, Pancreas, 2003, 26(1): 56-64
26 Ito A, Koshikawa N, Mochizuki S. Hypoxia-inducible factor-1 mediates the expression of DNA polymerase iota in human tumor cells. Biochem Biophys Res Commun. 2006,351(1): 306-311
27 Yasuda S, Arii S, Mori A, Isobe N, et al. Hexokinse Ⅱ and VEGF expression in liver tumors: correlation with hypoxia-inducible factor 1 alpha and its significance[J] J Hepatol. 2004,40(1):117-123
28 王燕,汪睿等.缺氧诱导因子-1 与肿瘤治疗研究进展. Medical Recapitulate 2004, 10(1): 5-9
29 Yang QC, Zeng BF, Shi ZM Inhibition of hypoxia-induced angiogenesis by trichostatin A via sup-pression of HIF-lα activity in human osteo sarcoma. J Exp Clin Cancer Res. 2006, 25(4):593-599
30 丁磊,陈孝平,王海平。肝癌及癌旁组织中 HIF-1α基因 蛋白的表达及其临床意义。肝胆外科杂志, 2004, 12(1):32-35
31 Semenza GL. O2-regulated gene expression: transcript- tional control of cardiorespiratory physiology by HIF-1[J]. J Appl Physiol,2004,96(3):1173-1177
32 Na X, Wu G Ryan CK, et al. Overproduction of vascular endothelial growth factor related to von Hippel- lindautumor suppressor gene mutations and hypoxia- inducible factor-1 alpha expression in renal cell carcinomas .J Urol, 2003, 170(2 Pt 1): 588-592
33 Lidgren A, Hedberg Y, Grankvist K ,et al. The expression of hypoxia-inducible factor 1alpha is a favorable independent prognostic factor in renal cell carcinoma. Clin Cancer Res, 2005, 11(3):1129-1135
34 Mizukami Y Li J, Zhang X, et al, Hypoxia-inducible factor-1 independent regulation of vascular endothelial growth factor by hypoxia in colon cancer. Cancer Res, 2004,64(5): 1765-1772
35 Bos R, van der Groep P, Greijer AE, et al. Levels of hypoxia-inducible factor 1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. Cancer, 2003, 97(6): 1573-1581
36 Sivridis E, Giatromanolaki A, Gatter KC, et al. Association of hypoxia-inducible factor 1alpha and 2alpha with activated angiogenic pathways and prognosis in patientswith endometrial carcinoma. Cancer, 2002, 95(5): 1055-1063
37 Koukourakis MI, Giatromanolaki A, Sivridis E, et al. Hypoxia-inducible factor (HIF1αand HIF2α), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer. Int J Radiat Oncol Biol Phys, 2002, 53(5): 1192-1202
38 Nakayama K, Kanzaki A, Hata K, et al. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) gene expression in human ovarian carcinoma. Cancer Lett, 2002, 176(2): 215-223
39 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(11): 1705-1708
40 Lee CH, Lee MK, Kang CD, et al. Differential expression of hypoxia-inducible factor 1 alpha and tumor cell proliferation between squamous cell carcinomas and adenocarcinomas among operable non small cell lung carcinomas[J]. J Korean Med Sei, 2003, 18(2): 196-203
41 王劲红,李福琴,艾丽敏等.宫颈癌中缺氧诱导因子-1α 的表 达 和 血 管 生 成 的 关 系 . 中 国 肿 瘤 与 临 床 , 2004, 24:1426-1427
42 Yu EZ, Li YY, Liu XH, et al, Antiapoptotic action of hypoxia-inducible factor-1 alpha in human endothelial cells [J]. Lab invest 2004, 84950:553-516
43 王晗,韩忠朝. 缺氧诱导因子-1 与细胞凋亡[J]. 国外医学生理、病理科学与临床分册,2005,1(15):52-55
44 Sasabe E, Tatemoto Y, Li D, et al. Mechanism of HIF-l alpha dependen Suppression of hypoxia-induced apoptosis in squamous cell carcinoma cells[J]. Cancer Sci, 2005, 96(7):394-402
45 Kim JY, Ahn HJ, Ryu JH, et al. BH3-only protein noxa is a mediator of hypoxic cell death induced by hypoxia- inducible factor 1 alpha [J]. J Exp Med, 2004, 199(1): 113-124
46 Lee MI, Kim JY, Suk K, et al. Identification of the hypoxia-inducible factor 1 alpha-responsive HGTD-P gene as a mediator in mitochondrial apoptotic pathway [J]. Mol Cell Biol, 2004, 24(9):3918-3127
47 Semenza GL. Targeting HIF-1 for cancer therapy[J]. Nat Rev Cancer, 2003, 3(10): 721-732
48 Xia S,Yu SY, Yuan XL,et al. Effects of hypoxia on expression of P-glycoprotein and multidrug resistance Protein in human lung adenocarcinoma A549 ecll line[J]. Chin Mde J,2004,84(8):663- 666
49 Wartenberg M, Ling FC,Muschen M,et al. Regulation of the multidrug resistance transporter P-glycoprotein in multicellular tumor spheroids by hypoxia-inducible factor 1(HIF-1)and reactive oxygen species[J]. FASEB J, 2003, 17(3): 503-505
50 Thorpe P. Antibody directed targeting of tumor vasculature.Proc Am Assoc Cancer Res, 1996, 37: 668
51 Mahendra G, KumarS, IsayevaT, et al. Antiangiogenic cancer gene therapy by adeno-associated virus 2-mediated stable expression of the soluble FMS-like tyrosine kinase-1 receptor. Cancer Gene Ther, 2004 Sep 10
52 Qian DZ, Wang X, Kachlmp SK, et al. The Histone Deacetylase Inhibitor NVP_LAQ824 Inhibits Angiogenesis and Has a Greater Antitumor Effect in Combination with the Vascular Endothelial Growth Factor Receptor Tyro-sine Kinase Inhibitor PTK787/ ZK222584. CancerRes, 2004, 64(18):6626-6634
53 Huckle WR, Roche RI. Post-transcriptional control of expression of sFlt-1, an endogenous inhibitor of vascular endothelial growth factor. J Cell Biochem, 2004, 93(1):120-132
54 Welsh SJ, Williams R, Kirkpatrick L, et al. Antitumor activity and pharmacodynamic properties of PX-478, an inhibitor of hypoxia-inducible factor-1 alpha. Mol Cancer Ther. 2004, 3(3):233-244
55 Mackenzie ED, Selak MA, Tennant DA, Cell permeating {alpha)-ketoglutarate derivatives alleviate pseudo-hypoxia in SDH deficient cells. Mol Cell Biol. 2007, 17(32):41-50
56 Rapisarda A, Aalek J, Hollinggshead M, et al. Schedule dependent inhibition of hypoxia-inducible factor-l alpha protein accumulation, angiogenesis, and tumor growth by topotecan in U251-HRE glioblastoma xenogragts. CancerRes, 2004, 64(19):6845-6848
57 Stoeltzing O, McCasrty MF, Wey JS, et al. Role of hypoxia-inducible factor l alpha in gastric cancer cell growth, angiogenesis, and vessel maturation. J Natl Cancer I, 2004, 96(12): 946-956
58 Yeo EJ, Chun YS, Cho YS, et al. YC-1: a potential anticancer drug targeting hypoxia-inducible factor 1. J Natl Cancer Inst 2003; 95: 516-525
59 Asc G, Xu X, Chu C, et al. Prognosic significance of erythropoieth expression in human endometrial carcinoma. Cancer, 2004, 100(11): 2376-2386
60 董典宁、孙平、智绪亭, 等, 反义 HIF-1α 基因治疗人肝癌裸鼠移植瘤的实验研究. 中国普通外科杂志,2006, 15(7):520-524
61 丁磊,陈孝平,王海平。封闭缺氧诱导因子作用位点对人肝癌细胞生长抑制的研究。华中科技大学学报(医学版),2005, 34(6):751-753
62 Li L, Lin X, Staver M, Evaluating hypoxia-inducible factor-l alpha as a cancer therapeutic target via inducible RNA interference in vivo. Cancer Res. 2005, 65(16): 7249-7258