發(fā)布時(shí)間：2018-04-30 12:20

  本文選題：BGC-823細(xì)胞 + 缺氧誘導(dǎo)因子��； 參考：《新疆醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:檢測(cè)HIF-1α、HIF-2α、GLUT1、PKM2在胃癌BGC-823細(xì)胞中的表達(dá)情況,研究siRNA技術(shù)特異性沉默人胃癌BGC-823細(xì)胞中缺氧誘導(dǎo)因子(HIF-1α、HIF-2α)后葡萄糖載體蛋白-1(GLUT1)、M2型丙酮酸激酶(PKM2)的蛋白及mRNA表達(dá)情況,探索HIF-1α、HIF-2α對(duì)胃癌細(xì)胞內(nèi)GLUT1、PKM2的調(diào)控作用。方法:實(shí)驗(yàn)總共分為4組,分別是對(duì)照組、轉(zhuǎn)染Control siRNA組、轉(zhuǎn)染HIF-1αsiRNA組、轉(zhuǎn)染HIF-2αsiRNA組,利用實(shí)時(shí)熒光定量RT-PCR法及Western-blot法檢測(cè)轉(zhuǎn)染HIF-1αsiRNA、HIF-2αsiRNA及Control siRNA后細(xì)胞內(nèi)GLUT1、PKM2蛋白及mRNA的表達(dá)情況。結(jié)果:利用RNA干擾技術(shù)能有效沉默BGC-823細(xì)胞中的HIF-1α、HIF-2α蛋白及mRNA的表達(dá)。轉(zhuǎn)染HIF-1αsiRNA、HIF-2αsiRNA后細(xì)胞內(nèi)GLUT1、PKM2在mRNA及蛋白表達(dá)水平均下降(P0.05),其中siRN A抑制HIF-1α后PKM2下調(diào)較siRNA抑制HIF-2α更明顯(P0.05),HIF-1α、HIF-2α對(duì)GLUT1的調(diào)控比較差異結(jié)果無明顯統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:研究結(jié)果提示HIF-1α和HIF-2α均可在轉(zhuǎn)錄及翻譯水平參與調(diào)控GLUT1、PKM2的表達(dá),其中HIF-1α對(duì)PKM2起到主要調(diào)控作用,HIF-1α、HIF-2α與GLUT1、PKM2共同作用,在促進(jìn)胃癌細(xì)胞的Warburg效應(yīng)和促進(jìn)腫瘤惡性表型方面起重要作用。
[Abstract]:Objective: to detect the expression of HIF-1 偽 -HIF-2 偽 -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 偽 (HIF-1 偽 HIF-2 偽) by siRNA technique. To investigate the regulatory effect of HIF-1 偽 -HIF-2 偽 on intracellular GLUT1 PKM2 in gastric cancer cells. Methods: the experiment was divided into four groups: control group, Control siRNA group, HIF-1 偽 siRNA group and HIF-2 偽 siRNA group. The expression of GLUT1PKM2 protein and mRNA in the transfected HIF-1 偽 siRNAHIF-2 偽 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 偽 -HIF-2 偽 and mRNA in BGC-823 cells could be effectively silenced by RNA interference technique. After transfection of HIF-1 偽 siRNA-HIF-2 偽 siRNA, the expression level of GLUT1PKM2 in mRNA and protein was decreased. The down-regulation of PKM2 after siRN A inhibition of HIF-1 偽 was more obvious than that of siRNA inhibition of HIF-2 偽. There was no significant difference in the regulation of GLUT1 between siRN A and siRNA. Conclusion: both HIF-1 偽 and HIF-2 偽 are involved in the regulation of GLUT1PKM2 expression at the transcriptional and translation levels, and HIF-1 偽 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.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

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本文編號(hào)：1824534