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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。
Objective To investigate the effect of neuropilin 1 (NRP1) gene RNA interference plasmid on proliferation and apoptosis of gastric adenocarcinoma cell line SGC7901. Methods NRP1 gene-specific shRNA expression plasmid was constructed and transfected into human gastric adenocarcinoma SGC7901 cells by liposome. The expression of NRP1 protein was detected by Western blot. The proliferation and apoptosis of cells were detected by flow cytometry and Annexin-V-FITC / PI. Results After restriction analysis and sequencing, the shRNA-NRP1 plasmid was constructed correctly. After transfection of SGC7901 cells for 48 h, the expression of NRP1 protein was effectively inhibited. The percentage of cells in G0 / G1 phase was significantly increased, the number of S phase cells was significantly decreased and apoptosis Rate increased. Conclusion shRNA-NRP1 plasmid can effectively inhibit the expression of NRP1 protein in gastric adenocarcinoma SGC7901 cell line and arrest the cell cycle in G0 / G1 phase, thereby reducing the cell proliferation and inducing the cell apoptosis.