干扰PGI调控HIF-1α的表达以及促进乳腺癌细胞凋亡

发布时间：2018-12-10 22:23

【摘要】：本实验探究乳腺癌MCF-7细胞中干扰PGI的表达对HIF-1α的调控以及对细胞凋亡的影响。首先,使用慢病毒质粒转染构建干扰PGI(PGI-si RNA)的稳转细胞株,然后使用Hoechst33258染色和流式细胞术(FCM)检测细胞的凋亡,随后,采用RT-PCR检测PGI、HIF-1α和LDHA基因的表达,Western blotting检测Bcl-2、Bax、PGI、LDHA、HIF-1α和ERK蛋白的表达。成功构建PGI-si RNA的MCF-7稳转细胞株后,Hoechst33258染色实验结果显示,PGI-si RNA稳转细胞株中出现核固缩、染色质凝集等形态学改变,FCM结果亦表明PGI-si RNA稳转细胞株发生细胞凋亡。RT-PCR结果显示,PGI-si RNA稳转细胞株中PGI、LDHA和HIF-1α基因表达下调;Western blotting结果显示,PGI-si RNA稳转细胞株中LDHA、HIF-1α、ERK和Bcl-2蛋白表达下调,Bax蛋白表达上调。以上实验结果表明:在乳腺癌MCF-7细胞中,干扰PGI可抑制HIF-1α的表达,同时促进MCF-7细胞凋亡。
[Abstract]:The aim of this study was to investigate the effect of interfering PGI expression on HIF-1 伪 and apoptosis in breast cancer MCF-7 cells. First, the stable cell lines interfering with PGI (PGI-si RNA) were constructed by using lentivirus plasmid transfection, then apoptosis was detected by Hoechst33258 staining and flow cytometry (FCM), and then the expression of PGI,HIF-1 伪 and LDHA genes was detected by RT-PCR. The expression of Bcl-2,Bax,PGI,LDHA,HIF-1 伪 and ERK protein was detected by Western blotting. The results of Hoechst33258 staining showed that nuclear pyknosis, chromatin agglutination and other morphological changes were observed in PGI-si RNA stable transformed cell lines after the successful construction of MCF-7 stably transformed cell lines of PGI-si RNA. FCM results also showed that apoptosis occurred in PGI-si RNA stable cell lines. RT-PCR showed that the expression of PGI,LDHA and HIF-1 伪 genes were down-regulated in PGI-si RNA stably transformed cell lines. Western blotting results showed that the expression of LDHA,HIF-1 伪, ERK and Bcl-2 protein was down-regulated and Bax protein was up-regulated in PGI-si RNA stable cell line. The results showed that interfering with PGI could inhibit the expression of HIF-1 伪 and promote the apoptosis of MCF-7 cells in breast cancer MCF-7 cells.
【作者单位】：重庆医科大学干细胞与组织工程研究室重庆医科大学;
【基金】：国家自然科学基金(No.31271368)资助
【分类号】：R737.9

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