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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 study the regulatory effect of miR-142-3p lentiviral vector on TET2 and its effect on the proliferation of SKOV3 cells. Methods: The pSicoR-miR-142-3p and pMIR-Report-TET2 overexpression vectors were constructed and packaged with recombinant lentivirus. Real-time quantitative PCR, Western blot and dual luciferase reporter assay were used to validate miR-142-3p and TET2 targeting role of the application of MTT assay of miR-142-3p lentivirus particles on SKOV3 cell proliferation. Results: pSicoR-miR-142-3p and pMIR-Report-TET2 recombinant plasmids and lentivirus particles that can carry miR-142-3p were successfully constructed. The overexpression of miR-142-3p significantly inhibited the expression of TET2 protein and mRNA (P <0.05), while the inhibition of miR-142-3p expression significantly increased the expression of TET2 protein and mRNA (P <0.05). MTT assay confirmed that miR- -3p significantly inhibited SKOV3 cell proliferation. Conclusion: Overexpression of miR-142-3p can effectively inhibit the expression of TET2 mRNA and protein in SKOV3 cells and inhibit the proliferation of SKOV3 cells.