RhoC慢病毒干扰载体的构建及其对卵巢癌SKOV3细胞中RhoC mRNA表达的影响  

Construction of RhoC lentivirus interference vector and its effect on expression of RhoC mRNA in ovarian cancer SKOV3 cells

作　　者：王珂[1] 邹积艳[1] 汪丽[2] 杜珍武[3] 王俊容[1] 叶聪[1] 潘颖[1] 

机构地区：[1]吉林大学中日联谊医院妇产科,吉林长春130033 [2]吉林大学中日联谊医院研究生管理办公室,吉林长春130033 [3]吉林大学第二医院中心实验室,吉林长春130041

出　　处：《吉林大学学报（医学版）》2015年第2期304-306,I0004,I0005,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(20090747)

摘　　要：目的:构建慢病毒介导的能够稳定沉默卵巢癌SKOV3细胞中RhoC基因的干扰载体,观察其对卵巢癌SKOV3细胞中RhoC基因表达的抑制作用。方法:人卵巢癌SKOV3细胞分为psiHIV-RhoC1组(转染Lenti-shRhoC1)、psiHIV-RhoC2组(转染Lenti-shRhoC2)、阴性对照组(转染Lenti-NC)和空白对照组(未经转染)。观察各组SKOV3细胞中绿色荧光蛋白的表达;采用RT-PCR法检测各组SKOV3细胞中RhoC mRNA的表达水平。结果:psiHIV-RhoC1、psiHIV-RhoC2和阴性对照组SKOV3细胞中均可见绿色荧光蛋白的表达。成功构建了特异性干扰载体,将其转染至SKOV3细胞后,psiHIV-RhoC1和psiHIV-RhoC2组SKOV3细胞中RhoC mRNA的表达水平较空白对照组明显降低。结论:构建的重组慢病毒RhoC shRNA干扰载体能有效抑制SKOV3细胞中RhoC mRNA的表达。Objective To construct the interference vector stably silencing Rhoc gene in the ovarian cancer SKOV3 cells induced by lentivirus,and to observe its inbibitory effect on the expression of RhoC gene in the ovary cancer SKOV3 cells.Methods The ovary cancer SKOV3 cells were divided into psiHIV-RhoC1group（transfected with Lenti-shRhoC1）,psiHIV-RhoC2group（transfected with Lenti-shRhoC2）,negative control group（transfected with Lenti-NC）and blank control group（with no transfection）.The expressions of green fluorescence protein in the SKOV3 cells in various groups were detected;the expression levels of RhoC mRNA in the SKOV3 cells in various groups were detected by RT-PCR method.Results There were the expressions of the green fluorescence protein in the SKOV3 cells in psiHIV-RhoC1,psiHIV-RhoC2,and negative control groups. The specific interference vector was constructed and transfected into SKOV3 cells successfully.Compared with bank control group,the expression levels of RhoC mRNA in the SKOV3 cells in psiHIV-RhoC1,psiHIV-RhoC2,and negative control groups were decreased.Conclusion The recombinant lentivirus RhoC shRNA interference vector can suppress the expression of RhoC mRNA in SKOV3 cells effectively.

关 键 词：RNA干扰 慢病毒 卵巢肿瘤 RHOC基因 

分 类 号：R711.75[医药卫生—妇产科学]