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

• 基础研究 • 上一篇    下一篇

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. 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. 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: 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