细胞周期蛋白依赖性激酶7基因沉默或敲除抑制卵巢癌细胞增殖及其机制探讨

发布时间：2018-06-15 05:46

  本文选题：卵巢肿瘤 + 细胞增殖　； 参考：《肿瘤》2017年11期

【摘要】：目的 :探讨细胞周期蛋白依赖性激酶7(cyclin-dependent kinase 7,CDK7)对卵巢癌细胞增殖的作用。方法:应用特异性siRNA转染法沉默卵巢癌OVCA433、TOV-112D和IGROV1细胞中CDK 7基因表达后,采用CCK-8法检测细胞增殖能力。应用成簇的规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeat,CRISPR)及其相关核酸内切酶9(CRISPR-associated endonuclease 9,Cas9)基因编辑系统敲除卵巢癌HEY、OVCA420、OVCA433和IGROV1细胞中的CDK 7基因后,采用克隆形成实验检测细胞克隆形成能力。用CDK7抑制剂THZ1处理卵巢癌TOV-112D、IGROV1、OVCA433、OVCAR8、OV90、SKOV3和COV413B细胞后,采用CCK-8法和克隆形成实验检测TOV-112D、IGROV1、OVCA433、OVCAR8和OV90细胞增殖和克隆形成能力,蛋白质印迹法检测IGROV1、OVCA433、SKOV3和COV413B细胞中CDK7蛋白表达水平和RNA聚合酶Ⅱ磷酸化水平。结果:沉默或敲除CDK 7基因后,卵巢癌细胞的增殖和克隆形成能力均明显减弱(P值均0.05)。THZ1处理可抑制卵巢癌细胞的增殖和克隆形成,并下调CDK7蛋白表达和RNA聚合酶Ⅱ磷酸化(P值均0.05)。结论:CDK7可能通过调控RNA聚合酶Ⅱ磷酸化,促进卵巢癌细胞的增殖。
[Abstract]:Aim: to investigate the effect of cyclin dependent kinase (7(cyclin-dependent kinase 7) CDK7 on the proliferation of ovarian cancer cells. Methods: the expression of CDK7 gene in OVCA433TOV-112D and IGROV1 cells was silenced by specific siRNA transfection, and the proliferation of OVCA433TOV-112D and IGROV1 cells was detected by CCK-8 method. The CDK7 gene of OVCA420OVCA433 and IGROV1 cells of ovarian cancer was knocked out by cluster regular interval short palindrome repeat regularly interspaced short palindromic repeat CRISPRR and its associated endonuclease 9 CRISPR-associated endonuclease 9 cas9 gene editing system. Clone formation assay was used to detect the ability of cell clone formation. The proliferation and clone formation of OVCA433OVCA8 OVCAR90OV90 SKOV3 and COV413B cells were detected by CCK-8 method and clone formation assay after treated with THZ1, a CDK7 inhibitor, and OVCA433OVCAR8 and OV90 cells were treated with THZ1, a CDK7 inhibitor, respectively, and the proliferation and cloning ability of OVCA433OVCAR8 and OV90 cells were detected by using CCK-8 assay and clone forming assay. The expression of CDK7 protein and the phosphorylation of RNA polymerase 鈪，

本文编号：2020925