缺氧对宫颈癌Hela细胞缺氧诱导因子-1α、葡萄糖转运蛋白-1表达的影响及与细胞增殖的关系

摘要

Objective To investigate the change of expression of hypoxia-inducible factor la (HIF-lα) and glucose transporter-1 (GLUT-1) in cervical cancer Hela cell line under hypoxia environment, and to explore the influence of hypoxia environment upon the expression of HIF-la and GLUT-1 and the cell proliferation in vitro. Methods Hypoxia environment of Hela cell line was induced by CoCl2- MTT assay was used to monitor the cell proliferation in different time and situation, to test the effect of CoCl2-induced hypoxia on the growth activity of human cervicalcancer Hela cell line in vitro. Reverse transcription polymerase chain reaction analysis and im-munocytochemistry were used to examine the changes of HIF-la and GLUT-I in human cervical cancer Hela cell line in vitro under hypoxia in different time. Results The growth activity of human cervical cancer Hela cell line was enhanced when cells were cultured in 50 /imol/L and 100 /xmol/L CoC12 for 24 hours (P <0.05). The proliferation was inhibited by higher concentration of CoC12 (P <0.01). The mRNA expression of HIF-laafter hypoxia exposure for 72 h was higher than that of control group (P <0.01). The mRNA expression of GLUT-1 was significantly increased after hypoxia exposure (P <0.05). Immunofluorescence analysis results showed that both the HIF-la protein and the GLUT-1 protein were increased in hypoxia conditions compared with the normoxia control group (P <0.01). Conclusion Acute or mild hypoxia conditions may promote the proliferation of human cervical cancer Hela cell line. Hypoxia conditions can up-regulate the expressions of HIF-la and GLUT-1 in human cervical cancer Hela cell line and lead to hypoxia tolerance of Hela. The fact that the expression of GLUT-1 protein elevated later than HIF-la protein suggests GLUT-1 gene and protein expression are up-regulated by HIF-la protein.%目的 通过动态观察缺氧时宫颈癌Hela细胞增殖状况及缺氧诱导因子-1α(HIF-1α)、葡萄糖转运蛋白-1(GLUT-1)表达的变化,探讨缺氧对HIF-1α及GLUT-1表达的影响及其与细胞增殖的关系.方法 使用CoCl2人工模拟缺氧环境,四甲基偶氮唑蓝比色法监测缺氧不同程度下不同时间点细胞增殖状况,观察缺氧对Hela细胞生长的影响,实时荧光逆转录聚合酶链反应(QRT-PCR)及细胞免疫荧光共聚焦显微镜检测缺氧后不同时间点HIF-1α及GLUT-1 mRNA及蛋白表达的变化.结果 50 μmol/L和100 μmol/L CoCl2作用24 h,Hela细胞增殖力均明显高于对照组(P＜0.05)；150、200、250 μmol/LCoCl2作用下,Hela细胞增殖力均低于对照组(P＜0.01)；缺氧72 h,HIF-1α mRNA表达量较对照升高(P＜0.01),缺氧24、48、72 h,GLUT-1 mRNA表达量均较对照明显升高(P＜0.05)；缺氧24、48、72h,Hela细胞HIF-1α及GLUT-1蛋白表达量均较对照组升高(P＜0.01).结论 轻度缺氧在一定时期内可促进Hela细胞增殖；缺氧能上调HIF-1α、GLUT-1基因及蛋白在Hela细胞中的表达,使之进一步耐受缺氧；缺氧主要通过上调HIF-1α蛋白表达,介导GLUT-1基因及后续蛋白表达.