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首页> 外文期刊>Journal of Huazhong University of Science and Technology >Effects of shRNA Targeting VEGF on VEGF mRNA Expression in Gastric Cancer Cells
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Effects of shRNA Targeting VEGF on VEGF mRNA Expression in Gastric Cancer Cells

机译:靶向VEGF的shRNA对胃癌细胞VEGF mRNA表达的影响

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摘要

In order to investigate the inhibitory effect of plasmid-mediated short hairpin RNA targeting vascular endothelial growth factor (VEGF) on the expression of VEGF mRNA in human gastric cancer cells, a plasmid vector for transcribing specific short hairpin RNA targeting VEGF (pU6-VEGF) was constructed, and then transfected into human gastric cancer cells using Lipofec-tamine2000. The VEGF mRNA expression level was detected by RT-PCR. RPMI1640 was used for blank control, and pSilencer 1.0-U6 empty plasmid for the negative control. Results showed the clone and sequence analysis revealed that the recombinant plasmid vector of pU6-VEGF was successfully constructed. The VEGF mRNA expression levels in blank control group, experimental group (pU6-VEGF) and negative control group (pSilencer1.0-U6) were 100%, 49% and 94%, respectively, indicating VEGF mRNA expression in the cells transfected with pU-VEGF vector was inhibited significantly as compared with blank control group and negative control group. It was concluded that the short hairpin RNA could significantly inhibit the expression of VEGF mRNA, which provided an experimental basis for treating human cancer with anti-angiogenesis.
机译:为了研究靶向于血管内皮生长因子(VEGF)的质粒介导的短发夹RNA对人胃癌细胞中VEGF mRNA表达的抑制作用,用于转录靶向VEGF的特异性短发夹RNA(pU6-VEGF)的质粒载体进行构建,然后使用Lipofec-tamine2000将其转染到人胃癌细胞中。通过RT-PCR检测VEGF mRNA的表达水平。 RPMI1640用于空白对照,pSilencer 1.0-U6空质粒用于阴性对照。结果表明,克隆和序列分析表明已成功构建pU6-VEGF重组质粒载体。空白对照组,实验组(pU6-VEGF)和阴性对照组(pSilencer1.0-U6)的VEGF mRNA表达水平分别为100%,49%和94%,表明在pU转染的细胞中VEGF mRNA的表达。与空白对照组和阴性对照组相比,-VEGF载体被显着抑制。结论:短发夹RNA可显着抑制VEGF mRNA的表达,为抗人血管生成治疗人类癌症提供了实验依据。

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