RNA干涉对人胃癌细胞端粒酶活性影响的研究

Study on Influence of RNA Interference in Activation of Telomerase Activity in Stomach Tumor Cell

目的:研究RNA干涉技术对人胃癌细胞株MGC80-3和BGC-823端粒酶活性表达的影响,以找到抑制端粒酶活性的简便方法。方法:根据RNA干涉原理,针对端粒酶基因序列设计了抑制端粒酶表达的核苷酸序列,克隆进真核表达载体中;并以脂质体转入到胃癌细胞系的MGC80-3和BGC-823细胞中,以空质粒为对照,48 h后收集细胞的总RNA,逆转录成cDNA后,通过荧光差异显示法检测基因表达的变化。结果:检测到一些基因的表达发生改变,端粒酶活性明显下降。结论:可以通过RNA干涉影响端粒酶活性。

Objective ： To detect the alteration of genes expression after RNA interference of Telomerase. Methods： According to the principle of RNA interference, a nucleotide sequence was designed to inhibit telomerase gene expression, the sequence was cloned into a eukaryotic vector and transferred into the stomach tumor cell lines, MGC80-3 and BGC-823 cells, the primary vector as a control, after 48 hours, the total RNA of each test was extracted and was synthesized to cDNA, the alteration of genes expression was detected by fluorescence differential display. Results： The alteration of genes expression was observed after the telomerase activity was inhibited. Conclusion ： The methods were effective in detection of the changes of a series of genes expression after telomerase RNA interference, it will be helpful to understand the relationships of the genes and telomerase, and the affection to cell growth.