闫杨, 付浩, 滑君, 张瑞秀, 陈丹琦, 孙开来, 孙秀菊. RNA干扰抑制RPS12基因表达对胃癌细胞凋亡的影响及分子机制探讨[J]. 中国肿瘤临床, 2008, 35(24): 1415-1418.
引用本文: 闫杨, 付浩, 滑君, 张瑞秀, 陈丹琦, 孙开来, 孙秀菊. RNA干扰抑制RPS12基因表达对胃癌细胞凋亡的影响及分子机制探讨[J]. 中国肿瘤临床, 2008, 35(24): 1415-1418.
YAN Yang, FU Hao, HUA Jun, ZHANG Rui-xiu, CHEN Dan-qi, SUN Kai-lai, SUN Xiu-ju. The Effects of Silencing the RPS12 Gene by RNAi on Apoptosis of Gastric Cancer Cells and the Possible Molecular Mechanism[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2008, 35(24): 1415-1418.
Citation: YAN Yang, FU Hao, HUA Jun, ZHANG Rui-xiu, CHEN Dan-qi, SUN Kai-lai, SUN Xiu-ju. The Effects of Silencing the RPS12 Gene by RNAi on Apoptosis of Gastric Cancer Cells and the Possible Molecular Mechanism[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2008, 35(24): 1415-1418.

RNA干扰抑制RPS12基因表达对胃癌细胞凋亡的影响及分子机制探讨

The Effects of Silencing the RPS12 Gene by RNAi on Apoptosis of Gastric Cancer Cells and the Possible Molecular Mechanism

  • 摘要: 目的: 探讨RPS12基因特异的RNA干扰对胃癌BGC823细胞凋亡的影响及机制。 方法: 设计并构建RPS12特异性shRNA表达载体(RPS12-shRNA),以Lipofectamine2000介导将RPS12特异性shRNA及Scram-ble-shRNA表达载体分别转染胃癌细胞。RT-PCR方法检测BGC823细胞中RPS12基因及凋亡相关基因Bcl-2和Bax mRNA的表达,流式细胞术(flow cytometry,FCM)分析RNA干扰对细胞凋亡的影响。 结果: RT-PCR结果显示,RPS12-shRNA表达载体转染第3、5、7天后,RPS12基因表达低于对照组,其中第5天下调最明显。RPS12特异性RNA干扰第5天、第7天后,Bcl-2 mRNA表达明显下调,BaxmRNA表达无明显改变。流式细胞术分析结果显示,RNA干扰RPS12基因第7天后,胃癌细胞凋亡率(17.84%)明显高于对照组(1.89%)。 结论: RNA干扰RPS12基因促进胃癌细胞凋亡作用可能是其作为上游基因通过下调Bcl-2基因表达而实现的。

     

    Abstract: Objective : To observe the influence of RPS12-specific RNA interference on the apoptosis of BGC823gastric cancer cells and to determine a possible mechanism. Methods : The RPS12-specific shRNA expres-sion vector was designed, constructed and transfected into gastric cancer cells using lipofectamine 2000.Scramble-shRNA was used as the control. The expression of RPS12 and apoptosis-associated genes Bcl-2and Bax was examined by RT-PCR. Apoptosis of gastric cancer cells was detected by flow cytometry. Results :RT-PCR showed that RPS12 mRNA expression was markedly downregulated on day 3, 5, and 7 after trans-fection of the RPS12-shRNA expression vector, especially on day 5. The expression of Bcl-2 mRNA was sig-nificantly downregulated on day 5 and 7 after transfection of the RPS12-shRNA vector. No changes werefound in Bax mRNA expression. On day 7 after transfection, the apoptosis rate of the transfected cells(17.84%) was higher than that of the control cells (1.89%). Conclusion : Suppression of the RPS12 gene withRNAi can increase apoptosis of gastric cancer cells, possibly by downregulating the upstream expression ofBcl-2.

     

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