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目的研究慢病毒介导短发夹RNA(shRNA)沉默血管生成素样蛋白4(ANGPTL4)基因对人宫颈癌Si Ha细胞增殖和凋亡的影响。方法用ANGPTL4 shRNA慢病毒感染Si Ha细胞,实时荧光定量PCR检测各组细胞ANGPTL4 mRNA水平,Western blot法检测ANGPTL4蛋白水平;MTT法和软琼脂克隆形成实验检测Si Ha细胞增殖能力;流式细胞术检测Si Ha细胞周期。藻红蛋白标记的annexin V和7-氨基放线菌素D联合染色(annexin V-PE/7-AAD)结合流式细胞术检测慢病毒感染后Si Ha细胞的凋亡。结果重组慢病毒感染Si Ha细胞后,LV3-ANGPTL4组Si Ha细胞中ANGPTL4 mRNA和蛋白表达水平降低;MTT结果显示,LV3-ANGPTL4组细胞增殖受抑制;软琼脂克隆形成实验显示,LV3-ANGPTL4组细胞克隆形成数减少;流式细胞术结果显示,LV3-ANGPTL4组G0/G1期细胞所占比例增加,细胞凋亡率增高。结论下调Si Ha细胞ANGPTL4的水平可抑制细胞增殖,使细胞周期阻滞于G0/G1期,诱导细胞凋亡。
Objective To investigate the effects of lentivirus-mediated short hairpin RNA (shRNA) silencing angiogenin-like protein 4 (ANGPTL4) on proliferation and apoptosis of human cervical carcinoma Si Ha cells. Methods ANGPTL4 shRNA lentivirus was used to infect Si Ha cells. ANGPTL4 mRNA level was detected by real-time fluorescence quantitative PCR. The ANGPTL4 protein level was detected by Western blot. The proliferation of Si Ha cells was detected by MTT assay and soft agar colony formation assay. Flow cytometry Si Ha cell cycle was examined. Phycoerythrin-labeled annexin V and annexin V-PE / 7-AAD combined with flow cytometry were used to detect Si ha cells apoptosis after lentivirus infection. Results The expression of ANGPTL4 mRNA and protein in Si3 + cells was decreased after infection with Si ha cells. MTT assay showed that the proliferation of cells in LV3-ANGPTL4 group was inhibited. The results of soft agar colony formation assay showed that LV3-ANGPTL4 group The results of flow cytometry showed that the proportion of cells in G0 / G1 phase increased and the apoptosis rate increased in LV3-ANGPTL4 group. Conclusion The down-regulation of ANGPTL4 in Si Ha cells can inhibit cell proliferation, arrest the cell cycle in G0 / G1 phase and induce apoptosis.