Adv-shRNA抑制NRP2表达对胃癌SGC7901细胞增殖的影响

Effect of Adv-shRNA inhibiting NRP2 expression on SGC7901 proliferation of gastric carcinoma

目的通过特异性抑制胃癌细胞株SGC7901中神经纤毛蛋白2(NRP2)基因的表达,检测RNA干扰片段对SGC7901细胞增殖能力的影响。方法将NRP2的特异性RNA干扰腺病毒载体转染至胃癌细胞SCG7901 72 h后荧光显微镜观察转染情况;采用RT-PCR法检测转染后NRP2基因mRNA表达情况;MTT法检测胃癌细胞增殖情况;Western印迹检测转染后NRP2蛋白表达情况。结果成功将NRP2-shRNA重组腺病毒载体转染至胃癌细胞SGC7901。各重组腺病毒转染组NRP2 mRNA水平均低于阴性对照组和空白组,其中以NRP2-shRNA-3抑制效应最强。阴性对照组与空白组细胞增殖迅速,两组间无显著差异(P>0.05),实验组经腺病毒载体转染后细胞增殖程度显著减少,与前两组相比有显著差异(P<0.05)。阴性对照组与空白组NRP2蛋白表达量明显高于实验组(P<0.01),而阴性对照组与空白组之间无显著差异(P>0.05)。结论重组腺病毒载体NRP2-shRNA可有效抑制胃癌细胞SCG7901中NRP2基因的mRNA和蛋白的表达,并在一定程度上抑制癌细胞增殖,可作为动物实验的理论基础和前期条件。

Objective To detect the influence of RNA interference fragment to ability of cell proliferation in SGC7901 cells with specific inhibition of NRP2 gene expression in gastric cancer cell line namely SGC7901. Methods NRP2 adenovirus vector was transfered into SCG7901 cells. After 72 hours, SCG7901 cells were observed, expression of mRNA in NRP2 gene was detected by RT-PCR ,proliferation of gastric cancer cell was detected by MTT and protein expression of NRP2 was detected after transfection by Western blot respectively. Results The transfection of NRP2-shRNA recombinant adenovirus vector into gastric cancer cell SGC7901 was successful. Level of NRP2 mRNA in transfection group was lower than that of blank and negative control groups and inhibitory effect of NRP2-shRNA-3 was the stron- gest. The cell proliferation was more rapidly in blank and negative control groups and there was no significant difference （ P 〉 0.05 ）. The cell proliferation was depressed in transfection group and there was significant difference compared to blank group or negative control group （ P 〈 0. 05 ）. Expression content in transfection group was higher than that of another two groups and there was significant difference （ P 〈 0.05 ） but there was no significant difference between blank and negative control groups （P 〉 0. 05）. Conclusions The expression of mRNA and protein of NRP2 in gastric cancer cell SGC7901 will be inhibited by NRP2-shRNA recombinant adenovirus vector and the same influence to cancer cell in some degree. It is theoretical basis and prophase conditions for animal experiment.