摘要
目的:研究miR-142-3p慢病毒载体对TET2的调控作用及其对SKOV3细胞增殖作用的影响。方法:构建pSicoR-miR-142-3p及pMIR-Report-TET2过表达载体,包装重组慢病毒,应用实时荧光定量PCR、Western blot及双荧光素酶报告基因系统检测方法验证miR-142-3p与TET2的靶向作用关系,应用MTT法检测miR-142-3p慢病毒颗粒对SKOV3细胞增殖的影响。结果:成功构建了pSicoR-miR-142-3p和pMIR-Report-TET2重组质粒,以及可携带miR-142-3p的慢病毒颗粒。miR-142-3p过表达明显抑制TET2蛋白及mRNA表达水平(P<0.05);抑制miR-142-3p表达则明显上调TET2蛋白及mRNA表达水平(P<0.05);MTT试验证实,miR-142-3p可显著抑制SKOV3细胞增殖。结论:过表达miR-142-3p可有效抑制SKOV3细胞中TET2 mRNA和蛋白水平表达,并抑制SKOV3细胞增殖。
Objective： To construct a recombinant lentivirus vector of miR-142-3p and verify its effect on ten-eleven translocation 2（ TET2） gene and the proliferation of SKOV3 cells.Methods： pSicoR-miR-142-3p、pMIR-Report-TET2 over-expression vector and the recombinant lentivirue vector were construsted. The targeted effect of miR-142-3p on TET2 gene was verified by real-time PCR test,Western blot and relative luciferase assay. The proliferation effects of miR-142-3p in SKOV3 cell lines were analyzed by MTT assay. Result： The recombinant plasmids of pSicoR-miR-142-3p and pMIR-Report-TET2 3 ＇-UTR were constructed successfully. Over-expression of miR-142-3p suppressed the mRNA and protein expression level of TET2 significantly（ P〈0. 05）. When the expression of miR-142-3p was suppressed,the expression levels of TET2 mRNA and protein were rose significantly（ P 〈0. 05）. MTT assay showed that the lentivirus vector of miR-142-3p could significantly inhibit the proliferation of SKOV3 cells（ P〈0. 05）. Conclusions： Over-expression of miR-142-3p could restrain the expression of the mRNA and protein level of TET2 gene in SKOV3 cells and effected on proliferation in human ovarian cancer cell line SKOV3
出处
《现代妇产科进展》
CSCD
北大核心
2016年第10期721-725,共5页
Progress in Obstetrics and Gynecology
基金
青年科学基金项目(No:31501049)
