慢病毒介导的VEGF-RNAi对宫颈癌细胞C33A凋亡的影响  

Effect of lentivirus-mediated VEGF-RNAi on apoptosis of cervical cancer cell line C33A

作　　者：李茸 王娟[1] LI Rong;WANG Juan(Department of Tumor Radiotherapy,First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院肿瘤放疗科,西安710061

出　　处：《山西医科大学学报》2022年第11期1402-1407,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81602676)。

摘　　要：目的构建针对血管内皮生长因子(VEGF)的慢病毒干扰载体,检测沉默VEGF基因后对宫颈癌细胞C33A凋亡的影响。方法通过对细胞感染复数(MOI)和病毒感染效率的测定,探索感染C33A细胞的最佳病毒浓度和促感染药物聚凝胺(Polybrene)的使用剂量,遂按照此最佳感染条件,将处于对数生长期的C33A细胞分为3组:空白对照组、阴性对照组(使用含有无意义链shRNA-scr的慢病毒载体感染)和实验组(使用含有VEGF-shRNA的慢病毒载体感染),然后检测C33A细胞中VEGF蛋白表达的差异,流式细胞术检测干扰后C33A细胞的凋亡情况。结果C33A细胞的慢病毒MOI为50,且使用5μg/ml的Polybrene时,慢病毒感染对C33A细胞无明显细胞毒性并达到最佳感染效果。成功感染慢病毒后,实验组C33A细胞VEGF蛋白的相对表达量较阴性对照组明显降低(23.3%±1.2%vs 96.7%±1.8%,P<0.05)。实验组C33A细胞凋亡率明显高于阴性对照组(46.03%±0.83%vs 15.26%±0.87%,P<0.05)。结论沉默宫颈癌细胞C33A的VEGF基因可下调VEGF蛋白的表达导致C33A细胞的凋亡。Objective To construct a lentivirus interference vector targeting VEGF and explore the effect of silencing VEGF gene on the proliferation of C33A cells.Methods Based on the multiplicity of infection(MOI)and the efficiency of virus infection,the optimal concentration of virus infecting C33A cells and the dosage of infection promoting drug(Polybrene)were screened.According to the best infection conditions,C33A cells in logarithmic growth phase were divided into three groups:blank control group,negative control group(transfected with lentiviral vector containing nonsense strand shRNA scr),experimental group(transfected with lentiviral vector containing VEGF shRNA).Then VEGF protein expression was detected in C33A cells,and the apoptosis of C33A cells was detected after interference in different groups by flow cytometry.Results When the MOI of lentivirus infection was 50 and the dosage of Polybrene was 5μg/ml,the lentivirus infection had no obvious cytotoxicity to C33A cells and the infection efficacy was the best.After successful infection with lentivirus,the relative expression of VEGF protein in experimental group was significantly lower than that in negative control group(23.3%±1.2%vs 96.7%±1.8%,P<0.05).The apoptosis rate in experimental group was significantly higher than that in negative control group(46.03%±0.83%vs 15.26%±0.87%,P<0.05).Conclusion Silencing of VEGF gene can induce the apoptosis of C33A cells by downregulating the expression of VEGF protein.

关 键 词：宫颈癌 慢病毒 血管内皮生长因子 细胞增殖 细胞凋亡 

分 类 号：R737.33[医药卫生—肿瘤]