shRNA干扰HIF-1α对人宫颈癌细胞黏附分子表达的影响  

Effect of shRNA interference targeting hypoxia inducible factor-1α on expression of adhesion molecules in human cervical carcinoma HeLa cells

作　　者：董兰兰[1] 袁响林[1] 于世英[1] 谢涛[1] 杨彬[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤科,武汉430030

出　　处：《中国肿瘤生物治疗杂志》2008年第1期46-50,共5页Chinese Journal of Cancer Biotherapy

基　　金：湖北省卫生厅重点资助项目(NoJX1A06)~~

摘　　要：目的:在细胞和移植瘤水平研究shRNA干扰低氧诱导因子-1α(hypoxia inducible factor-1,HIF-1α)对宫颈腺癌HeLa细胞中黏附分子CD44v6及上皮型钙黏素(E-cadherin)表达的影响。方法:将人宫颈癌Hela细胞株(记为H0细胞)、转染pGenesil-1空白质粒的HeLa细胞株(记为H1细胞)及转染HIF-1α-shRNA质粒的HeLa细胞株(记为H2细胞)分别行常氧培养及低氧(1%O2)培养,应用Western blotting检测每组HeLa细胞中HIF-1α、CD44v6、E-cadherin及β连接素(β-catenin)在细胞水平的表达情况,应用免疫组织化学法检测HIF-1α、CD44v6、E-cadherin及β-catenin在上述3种HeLa细胞裸鼠移植瘤模型瘤组织水平的表达情况。结果:Western blotting分析显示,H0和H1细胞低氧组检测到HIF-1α、β-catenin、CD44v6蛋白呈强阳性,而E-cadherin表达减弱;常氧状态下H0、H1和H2组及低氧下H2组HIF-1α无表达、E-cadherin表达呈阳性。免疫组化分析发现H0及H1组移植瘤组织中HIF-1α、E-cadherin、β-catenin及CD44v6的表达分别为0.651±0.057、0.198±0.015、0.465±0.061及0.687±0.102,0.671±0.069、0.201±0.020、0.452±0.038及0.701±0.071,H2组分别为0.194±0.023、0.548±0.045、0.421±0.052及0.187±0.057,H2组与H0和H1组间HIF-1α、E-cadherin、及CD44v6表达的差异有统计学意义(P<0.05);相关性分析提示HIF-1α与E-cadherin、CD44v6的表达有一定相关性。结论:通过shRNA干扰抑制人宫颈癌HeLa细胞HIF-1α,可一定程度上下调低氧环境中宫颈癌HeLa细胞黏附分子CD44v6及上调E-cadherin的表达。Objective： To investigate the effect of RNA interference （RNAi） targeting hypoxia inducible factor-1α（HIF-1α） on the expression of cell adhesion molecules （CD44v6, E-cadherin and β-catenin ） in human cervical carcinoma HeLa cells. Methods：Human cervical carcinoma He1a cell line （marked as H0 cell） , transfection pGenesil-1 Hela cell line （marked as H1 cell） and transfection HIF-1α-shRNA Hela cell line （marked as H2 cell） were subjected to hypoxia and normal cultivation for different time periods. The expression of HIF-1α, CD44v6, E-cadherin and β-catenin proteins were measured by Western blotting in each group. The expression of HIF-1α, CD44v6, E-cadherin and β-catenin in the above 3 groups were detected in transplantation tumors in nude mice by immunohistochemical method. Results： Western blotting showed that the expression of HIF-1α, β-catenin and CD44v6 protein was increased and the expression of E-cadherin were decreased in H0 and H1 groups under hypoxic condition. No HIF-1α expression were found in H0,H1 and H2 groups under normal condition and in H2 group under hypoxic condition. E-cadherin expression was noticed. Immunohistochemical study showed that the expression of HIF-1α, E-cadherin, β-catenin and CD44v6 in the H0 group were （0.651±0.057）, （0.198±0.015 ）, （0.465±0.061 ）, and （ 0.687±0.102）, respectively; those in the H1 group were （0.671±0.069） , （0.201±0.020）, （0.452±0.038）, and （0.701±0.071 ）, respectively; and those in the H2 group were （0.194±0.023 ） , （0.548±0.045 ）, （0.421±0.052 ）, and （0.187±0.057 ）, respectively. HIF-1α, E-cadherin, β-catenin and CD44v6 expression in H2 group were significantly different from those in the H0 and H1 groups （P 〈 0.05 ）. We also noticed that HIF-1α expression were correlated with E-cadherin and CD44v6 expression. Conclusion ： shRNA targeting HIF-1α in human cervical carcinoma HeLa cells can down-regulate CD44v6 and up-regulate E-cadherin expression un

关 键 词：低氧诱导因子-1Α 黏附分子 宫颈癌细胞 肿瘤转移 短发夹RNA 

分 类 号：R737.33[医药卫生—肿瘤] R730.54[医药卫生—临床医学]