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目的:探讨长链非编码RNA MEG3在胃癌组织及细胞系中的表达水平,以及过表达MEG3对胃癌细胞增殖能力的影响,并探索其可能的作用机制。方法:用定量反转录PCR(qRT-PCR)技术检测胃癌组织及细胞系中MEG3的表达水平;通过转染pcDNA-MEG3上调MEG3的表达水平,并通过qRT-PCR检测转染效率。用MTT和克隆形成试验检测上调MEG3的水平对BGC-823细胞和MGC-803细胞的增殖能力的影响,Western blot检测这两种细胞中上调MEG3的水平对p53蛋白的表达水平的影响。结果:相比正常胃组织及细胞,在胃癌组织和细胞中MEG3的表达出现显著下调,SGC7901、MGC-803和BGC-823细胞中MEG3的表达水平分别是正常胃上皮细胞GES-1中的73.6%、42.0%和27.1%(P<0.05)。BGC-823细胞和MGC-803细胞中转染pcDNA-MEG3能显著上调MEG3的表达,MEG3的表达水平分别为对照组的252倍和311倍(P<0.05);MTT和克隆形成试验显示,上调MEG3的表达能降低BGC-823细胞和MGC-803细胞的增殖能力。Western blot实验显示,转染了pcDNA-MEG3的MGC-803和BGC-823细胞中p53的表达相较对照组中显著增加。结论:胃癌组织及细胞中MEG3的表达下调,且这可能通过抑制p53蛋白的激活从而促进胃癌细胞增殖,影响胃癌的发生和发展。
Objective: To investigate the expression of long-chain non-coding RNA MEG3 in gastric cancer tissues and cell lines, as well as the effect of MEG3 overexpression on the proliferation of gastric cancer cells and to explore its possible mechanism. Methods: The expression of MEG3 in gastric cancer tissues and cell lines was detected by quantitative reverse transcription PCR (qRT-PCR). The expression of MEG3 was up-regulated by pcDNA-MEG3 transfection and the transfection efficiency was detected by qRT-PCR. The effect of MEG3 up-regulation on the proliferation of BGC-823 cells and MGC-803 cells was detected by MTT assay and clone formation assay. Western blot was used to detect the effect of MEG3 upregulation on p53 protein expression. Results: Compared with normal gastric tissues and cells, MEG3 expression was significantly down-regulated in gastric cancer tissues and cells. The expression levels of MEG3 in SGC7901, MGC-803 and BGC-823 cells were 73.6 %, 42.0% and 27.1% (P <0.05). MEG3 expression was up-regulated in BGC-823 cells and MGC-803 cells transfected with pcDNA-MEG3, the expression levels of MEG3 were 252-fold and 311-fold higher than that of the control group (P <0.05); MTT and clonogenic assay showed that up- MEG3 expression can reduce the proliferation of BGC-823 cells and MGC-803 cells. Western blot showed that the expression of p53 in MGC-803 and BGC-823 cells transfected with pcDNA-MEG3 was significantly increased compared with the control group. Conclusion: The expression of MEG3 in gastric cancer tissues and cells is down-regulated, and this may promote the proliferation of gastric cancer cells by inhibiting the activation of p53 protein, and affect the occurrence and development of gastric cancer.