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

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。

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

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-RhoC1 group(transfected with Lenti-shRhoC1),psiHIV-RhoC2 group(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.