吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (02): 304-306.doi: 10.13481/j.1671-587x.20150219

• 基础研究 • 上一篇    下一篇

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

王珂1, 邹积艳1, 汪丽2, 杜珍武3, 王俊容1, 叶聪1, 潘颖1   

  1. 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
  • 收稿日期:2014-02-12 出版日期:2015-03-28 发布日期:2015-04-04
  • 通讯作者: 潘颖, 主任医师, 硕士研究生导师(Tel:0431-84995253, E-mail:panying424@163.com) E-mail:panying424@163.com
  • 作者简介:王珂(1988-), 女, 吉林省长春市人, 住院医师, 医学硕士, 主要从事妇产科疾病实验方面的研究。
  • 基金资助:

    吉林省科技厅科研基金资助课题(20090747)

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

WANG Ke1, ZOU Jiyan1, WANG Li2, DU Zhenwu3, WANG Junrong1, YE Cong1, PAN Ying1   

  1. 1. Department of Gynecology and Obstetrics, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
    2. Graduate Management Office, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
    3. Central Laboratory, Second Hospital, Jilin University, Changchun 130041, China
  • Received:2014-02-12 Online:2015-03-28 Published:2015-04-04

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。

关键词: RNA干扰, 慢病毒, 卵巢肿瘤, RhoC基因

Abstract:

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.

Key words: RNA interference, lentivirus, ovary neoplasms, RhoC gene

中图分类号: 

  • R711.75