胃癌BGC-823细胞中HIF-1α、HIF-2α对GLUT1、PKM2的调控作用

发布时间：2018-04-30 12:20

本文选题：BGC-823细胞 + 缺氧诱导因子　；参考：《新疆医科大学》2017年硕士论文

【摘要】：目的:检测HIF-1α、HIF-2α、GLUT1、PKM2在胃癌BGC-823细胞中的表达情况,研究siRNA技术特异性沉默人胃癌BGC-823细胞中缺氧诱导因子(HIF-1α、HIF-2α)后葡萄糖载体蛋白-1(GLUT1)、M2型丙酮酸激酶(PKM2)的蛋白及mRNA表达情况,探索HIF-1α、HIF-2α对胃癌细胞内GLUT1、PKM2的调控作用。方法:实验总共分为4组,分别是对照组、转染Control siRNA组、转染HIF-1αsiRNA组、转染HIF-2αsiRNA组,利用实时荧光定量RT-PCR法及Western-blot法检测转染HIF-1αsiRNA、HIF-2αsiRNA及Control siRNA后细胞内GLUT1、PKM2蛋白及mRNA的表达情况。结果:利用RNA干扰技术能有效沉默BGC-823细胞中的HIF-1α、HIF-2α蛋白及mRNA的表达。转染HIF-1αsiRNA、HIF-2αsiRNA后细胞内GLUT1、PKM2在mRNA及蛋白表达水平均下降(P0.05),其中siRN A抑制HIF-1α后PKM2下调较siRNA抑制HIF-2α更明显(P0.05),HIF-1α、HIF-2α对GLUT1的调控比较差异结果无明显统计学意义(P0.05)。结论:研究结果提示HIF-1α和HIF-2α均可在转录及翻译水平参与调控GLUT1、PKM2的表达,其中HIF-1α对PKM2起到主要调控作用,HIF-1α、HIF-2α与GLUT1、PKM2共同作用,在促进胃癌细胞的Warburg效应和促进肿瘤恶性表型方面起重要作用。
[Abstract]:Objective: to detect the expression of HIF-1 伪 -HIF-2 伪 -GLUT1PKM2 in BGC-823 cells of gastric cancer, and to study the expression of glucose carrier protein -1 (GLUT1) pyruvate kinase M2 (PKM2) and the expression of mRNA in human gastric cancer BGC-823 cells after the specific silencing of HIF-1 伪 (HIF-1 伪 HIF-2 伪) by siRNA technique. To investigate the regulatory effect of HIF-1 伪 -HIF-2 伪 on intracellular GLUT1 PKM2 in gastric cancer cells. Methods: the experiment was divided into four groups: control group, Control siRNA group, HIF-1 伪 siRNA group and HIF-2 伪 siRNA group. The expression of GLUT1PKM2 protein and mRNA in the transfected HIF-1 伪 siRNAHIF-2 伪 siRNA and Control siRNA cells were detected by real-time fluorescence quantitative RT-PCR assay and Western-blot assay. Results: the expression of HIF-1 伪 -HIF-2 伪 and mRNA in BGC-823 cells could be effectively silenced by RNA interference technique. After transfection of HIF-1 伪 siRNA-HIF-2 伪 siRNA, the expression level of GLUT1PKM2 in mRNA and protein was decreased. The down-regulation of PKM2 after siRN A inhibition of HIF-1 伪 was more obvious than that of siRNA inhibition of HIF-2 伪. There was no significant difference in the regulation of GLUT1 between siRN A and siRNA. Conclusion: both HIF-1 伪 and HIF-2 伪 are involved in the regulation of GLUT1PKM2 expression at the transcriptional and translation levels, and HIF-1 伪 plays a major role in the regulation of PKM2. It plays an important role in promoting Warburg effect and malignant phenotype of gastric cancer cells.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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