RNAi沉默survivin和HIF-1α基因对胃癌BGC-823细胞体外增殖和凋亡的影响

doi:10.13481/j.1671-587x.20180411

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (04): 753-758.doi: 10.13481/j.1671-587x.20180411

RNAi沉默survivin和HIF-1α基因对胃癌BGC-823细胞体外增殖和凋亡的影响

王淼¹, 母润红², 李明成¹, 李洪杰²

1. 北华大学医学检验学院免疫教研室, 吉林吉林 132013;
2. 北华大学基础医学院免疫教研室, 吉林吉林 132013

收稿日期:2017-11-19 出版日期:2018-07-28 发布日期:2018-07-27
通讯作者: 母润红,副教授,硕士研究生导师(Tel:0432-64608107,E-mial:murunhong@126.com) E-mail:murunhong@126.com
作者简介:王淼(1976-),女,吉林省吉林市人,副教授,医学硕士,主要从事肿瘤分子生物学方面的研究。
基金资助:
吉林省教育厅科研基金资助课题（2014第202）；吉林省吉林市科技局科研基金资助课题（201339052）

Effects of silencing survivin and HIF-1α genes with RNAi on proliferation and apoptosis of human gastric cancer BGC-823 cells

WANG Miao¹, MU Runhong², LI Mingcheng¹, LI Hong Jie²

1. Department of Immunology, School of Laboratory Medicine, Beihua University Jilin 132013, China;
2. Department of Immunology, School of Basic Medical Sciences, Beihua University, Jilin 132013, China

Received:2017-11-19 Online:2018-07-28 Published:2018-07-27

摘要/Abstract

摘要： 目的：应用RNA干扰（RNAi）技术抑制胃癌BGC-823细胞中survivin和HIF-1α基因的表达，探讨其对胃癌BGC-823细胞增殖和凋亡的影响。方法：分别设计并合成经过化学修饰的靶向survivin和HIF1α mRNA的小干扰RNA （siRNAs），同时合成错义RNA （SCR），采用HifectinⅡ真核体外转染胃癌BGC-823细胞，将转染siRNA-survivin、siRNA-HIF-1α和SCR的各组细胞分别命名为sis组、siH组和SCR组，同时设空白对照组（无血清培养基），RT-PCR法检测各组细胞中survivin和HIF-1αmRNA表达水平。将胃癌BGC-823细胞分为单干扰组（survivin-siRNA，sis组）、联合干扰组（survivin-siRNA+HIF1α-siRNA，sis+siH组）、非靶向特异性组（SCR组）和空白对照组（无血清培养基），MTT法检测各组细胞增殖活性，Western blotting法检测各组细胞中survivin和HIF-1α蛋白表达水平，流式细胞术检测各组细胞凋亡率。结果：与空白对照组比较，sis组细胞中survivin mRNA表达水平和siH组细胞中HIF-1αmRNA表达水平明显降低（P<0.05或P<0.01）。与空白对照组比较，SCR组细胞增殖活性无明显变化（P>0.05），sis组和sis+siH组细胞增殖活性明显降低（P<0.05）；与sis组比较，sis+siH组细胞增殖活性明显降低（P<0.05）。与空白对照组比较，sis+siH组细胞中surviving和HIF-1α蛋白表达水平明显降低（P<0.05或P<0.01），细胞凋亡率明显升高（P<0.01）。结论：联合靶向沉默survivin和HIF-1α基因可下调靶基因的表达，抑制胃癌BGC-823细胞增殖，促进细胞凋亡，RNAi基因沉默技术有望成为治疗胃癌的新方法。

关键词: 小干扰RNA, 缺氧诱导因子1α, 人胃癌BGC-823细胞, 细胞增殖, survivin, 细胞凋亡

Abstract: Objective:To inhibit the expressions of survivin and HIF-1α using RNA interference(RNAi) technique,and to explore their effects on the proliferation and apoptosis of gastric cancer BGC-823 cells. Methods: Two pairs of siRNA were designed against the human survivin and HIF-1α separately.The human gastric cancer BGC-823 cells were transfected with siRNA-survivin,siRNA-HIF-1α and SCR and used as sis group,siH group,and scrambled siRNA (SCR) group;at the same time, blank control group(serum-free medium) was set up.RT-PCR was used to detect the expression levels of survivin and HIF-1α mRNA in the BGC-823 cells in various groups.The BGC-823 cells were transfected with siRNAs using HifectinⅡ.The BGC-823 cells were divided into sis group,sis+siH group,SCR group and blank control group.MTT assay was performed to detect the proliferation activity of cells.The expression levels of survivin and HIF-1α protein in the cells in various groups were detected by Western blotting method.Flow cytometry was used to detect the apoptotic rates of BGC-823 cells in various groups. Results:Compared with blank control group,the expression level of survivin mRNA in sis group was significantly desreased (P<0.05),and the expression level of HIF-1α mRNA in siH group was significantly desreased (P<0.05).Compared with blank control group,the proliferation ability of cells in SCR group had no significant change (P>0.05),and the proliferation abilities of cells in sis group and sis+siH group were significantly decreased(P<0.05);compared with sis group,the proliferation ability of cells in sis+siH group was significantly decreased(P<0.05).Compared with blank control group,the expression levels of survivin and HIF-1α in sis+siH group were significantly decreased (P<0.05 or P<0.01),and the apoptotic rates of BGC-823 cells were significantly increased(P<0.01). Conclusion:The combined targeting silencing surviving and HIF-1α genes can down-regulate the expressions of targeted genes,inhibit the proliferation of gastric cancer BGC-823 cells,and promote the apopotosis,indicating RNAi gene silencing may be a new method to treat gastric cancer.

Key words: human gastric cancer BGC-823 cells, hypoxia inducible factor 1α, apoptosis, cell proliferation, siRNA, survivin

中图分类号:

R735.2

王淼, 母润红, 李明成, 李洪杰. RNAi沉默survivin和HIF-1α基因对胃癌BGC-823细胞体外增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 753-758.

WANG Miao, MU Runhong, LI Mingcheng, LI Hong Jie. Effects of silencing survivin and HIF-1α genes with RNAi on proliferation and apoptosis of human gastric cancer BGC-823 cells[J]. Journal of Jilin University Medicine Edition, 2018, 44(04): 753-758.

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RNAi沉默survivin和HIF-1α基因对胃癌BGC-823细胞体外增殖和凋亡的影响

Effects of silencing survivin and HIF-1α genes with RNAi on proliferation and apoptosis of human gastric cancer BGC-823 cells

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