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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。
Objective: To investigate the effect of cyclin-dependent kinase 7 (CDK7) on the proliferation of ovarian cancer cells. Methods: The expression of CDK 7 gene in ovarian cancer OVCA433, TOV-112D and IGROV1 cells was silenced by specific siRNA. CCK-8 method was used to detect the cell proliferation. The ovarian cancers HEY, OVCA420 and OVCA433 were knocked down by using the clustered regularly interspaced short palindromic repeat (CRISPR) and its associated 9 (CRISPR-associated endonuclease 9) gene editing system. And CDK7 gene in IGROV1 cells, clone formation assay was used to examine cell clonality. After treatment of TOV-112D, IGROV1, OVCA433, OVCAR8, OV90, SKOV3 and COV413B cells with CDK7 inhibitor THZ1, the proliferation of TOV-112D, IGROV1, OVCA433, OVCAR8 and OV90 cells were detected by CCK- The ability of CDK7 protein expression and RNA polymerase II phosphorylation in IGROV1, OVCA433, SKOV3 and COV413B cells were detected by Western blotting. Results: The proliferation and clonogenic capacity of ovarian cancer cells were significantly decreased after silencing or knocking out CDK 7 gene (all P <0.05). THZ1 treatment inhibited the proliferation and colony formation of ovarian cancer cells and down-regulated the expression of CDK7 and phosphorylated RNA polymerase Ⅱ (all P <0.05). Conclusion: CDK7 may promote the proliferation of ovarian cancer cells by regulating the phosphorylation of RNA polymerase Ⅱ.