發(fā)布時間：2018-06-15 05:46

  本文選題：卵巢腫瘤 + 細(xì)胞增殖�。� 參考：《腫瘤》2017年11期

【摘要】：目的 :探討細(xì)胞周期蛋白依賴性激酶7(cyclin-dependent kinase 7,CDK7)對卵巢癌細(xì)胞增殖的作用。方法:應(yīng)用特異性siRNA轉(zhuǎn)染法沉默卵巢癌OVCA433、TOV-112D和IGROV1細(xì)胞中CDK 7基因表達(dá)后,采用CCK-8法檢測細(xì)胞增殖能力。應(yīng)用成簇的規(guī)律間隔短回文重復(fù)序列(clustered regularly interspaced short palindromic repeat,CRISPR)及其相關(guān)核酸內(nèi)切酶9(CRISPR-associated endonuclease 9,Cas9)基因編輯系統(tǒng)敲除卵巢癌HEY、OVCA420、OVCA433和IGROV1細(xì)胞中的CDK 7基因后,采用克隆形成實驗檢測細(xì)胞克隆形成能力。用CDK7抑制劑THZ1處理卵巢癌TOV-112D、IGROV1、OVCA433、OVCAR8、OV90、SKOV3和COV413B細(xì)胞后,采用CCK-8法和克隆形成實驗檢測TOV-112D、IGROV1、OVCA433、OVCAR8和OV90細(xì)胞增殖和克隆形成能力,蛋白質(zhì)印跡法檢測IGROV1、OVCA433、SKOV3和COV413B細(xì)胞中CDK7蛋白表達(dá)水平和RNA聚合酶Ⅱ磷酸化水平。結(jié)果:沉默或敲除CDK 7基因后,卵巢癌細(xì)胞的增殖和克隆形成能力均明顯減弱(P值均0.05)。THZ1處理可抑制卵巢癌細(xì)胞的增殖和克隆形成,并下調(diào)CDK7蛋白表達(dá)和RNA聚合酶Ⅱ磷酸化(P值均0.05)。結(jié)論:CDK7可能通過調(diào)控RNA聚合酶Ⅱ磷酸化,促進(jìn)卵巢癌細(xì)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，

本文編號：2020925