血管生成素样蛋白4(ANGPTL4)敲低抑制宫颈癌SiHa细胞增殖并促进其凋亡  

Knockdown of angiopoietin-like protein 4 inhibits proliferation and promotes apoptosis in cervical cancer Si Ha cells

作　　者：聂丹[1] 刘玲[1] 夏纪毅 汪春燕[1] 詹平[1] 毛熙光[1] 

机构地区：[1]泸州医学院附属医院妇科 [2]四川医科大学药物与功能性食品研究中心,四川泸州646000

出　　处：《细胞与分子免疫学杂志》2016年第4期488-492,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：四川省卫生厅科研课题(130374);泸州市科技计划项目[2013-S-48(15/30)];泸州医学院青年基金项目(2014QN-06)

摘　　要：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 effect of lentivirus-mediated shRNA silencing of angiopoietin-like protein 4（ ANGPTL4） on the proliferation and apoptosis of cervical cancer Si Ha cells. Methods The ANGPTL4 lentiviral vectors were used to transfect Si Ha cells. Real-time quantitative PCR（ qRT-PCR） and Western blotting were respectively used to detect ANGPTL4 expression at mRNA and protein levels. The proliferation ability of Si Ha cells after transfection was assessed by MTT assay and colony formation assay. The cell cycle was examined by flow cytometry. The annexin V-phycoerythrin /7-aminoactinomycin D（ annexin Ⅴ-PE /7-AAD） staining combined with flow cytometry was used to examine the effect of ANGPTL4 silence on the apoptosis of Si Ha cells. Results After the ANGPTL4 lentiviral vectors were transfected into Si Ha cells,qRT-PCR and Western blotting showed that the expression of ANGPTL4 mRNA and protein were significantly inhibited in LV3-ANGPTL4 group. The MTT assay showed that the proliferation ability of Si Ha cells in LV3-ANGPTL4 group was also inhibited. Colony formation assay revealed that the colony number in LV3-ANGPTL4 group was reduced. The cells in G0 / G1 phase and the apoptosis rate increased in LV3-ANGPTL4 group. Conclusion The lentivirus-mediated ANGPTL4 shRNA can inhibit the proliferation,induce the cell cycle arrest in G0 / G1 phase,and promote the apoptosis in Si Ha cells.

关 键 词：血管生成素样蛋白4 慢病毒 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤] R392-33[医药卫生—临床医学]