神经纤毛蛋白1基因RNA干扰质粒对胃腺癌细胞SGC7901增殖和凋亡的影响

目的探讨神经纤毛蛋白1(Neuropilin1,NRP1)基因RNA干扰质粒对胃腺癌细胞SGC7901增殖和凋亡的影响。方法构建NRP1基因特异性shRNA表达质粒,经脂质体介导转染入人胃腺癌SGC7901细胞。采用Western blot法检测细胞中NRP1蛋白的表达水平,应用流式细胞术和Annexin-V-FITC/PI法检测细胞的增殖水平和凋亡率。结果经酶切及测序鉴定证明,shRNA-NRP1质粒构建正确,转染SGC7901细胞48h后,能有效抑制NRP1蛋白的表达,G0/G1期细胞百分比显著升高,S期细胞显著减少,细胞凋亡率增加。结论 shRNA-NRP1质粒能有效抑制胃腺癌SGC7901细胞株NRP1蛋白的表达,细胞周期阻滞在G0/G1期,从而降低细胞的增殖水平,诱导细胞进入凋亡期。

Effect of Neuropilin1 Gene Specific RNA Interfering Plasmid on Proliferation and Apoptosis of Gastric Carcinoma Cells

Objective To investigate the effect of neuropilin1(NRP1)gene specific RNA interfering plasmid on the proliferation and apoptosis of gastric carcinoma cells.Methods NRP1 gene specific shRNA expression plasmid was constructed and transfected to human gastric carcinoma SGC7901 cells in mediation of liposome.The expression level of NRP1 in the cells was determined by Western blot,and the proliferation level and apoptosis rate of the cells by flow cytometry and Annexin-V-FITC/PI method.Results Both restriction analysis and sequencing proved that plasmid shRNA-NRP1 was constructed correctly.The expression of NRP1 in the SGC7901 cells 48 h after transfection was inhibited effectively.The percentage of cells at G0/G1 phase increased significantly,while those at S phase decreased significantly,and the apoptosis rate of cells increased.Conclusion Plasmid shRNA-NRP1 inhibited the expression of NRP1 in SGC7901 cells effectively and arrested the cell cycle at G0/G1 phase,thereby decreased the proliferation level and induced the apoptosis of the cells.