학술논문
沉默CDK7通过下调YAP表达诱导子宫内膜癌细胞凋亡 / Silencing CDK7 promotes the apoptosis of endometrial cancer cells by downregulating the expression of YAP
Document Type
Academic Journal
Author
Source
肿瘤药学 / Anti-Tumor Pharmacy. 13(6):729-734
Subject
Language
Chinese
ISSN
2095-1264
Abstract
目的 研究沉默细胞周期蛋白依赖性激酶7(CDK7)诱导Yes相关蛋白(YAP)表达下调对子宫内膜癌HEC-1-A细胞凋亡的影响.方法 利用siRNA干扰技术沉默HEC-1-A细胞中CDK7的表达,采用qPCR和Western blotting检测siRNA干扰后CDK7的表达水平,CCK-8实验检测沉默CDK7对HEC-1-A细胞活力的影响,流式细胞术检测沉默CDK7对HEC-1-A细胞凋亡的影响,Western blotting检测沉默CDK7对YAP和磷酸化YAP蛋白及其下游蛋白Cyr16、CTGF表达的影响,凋亡实验检测共转染si-CDK7和pcDNA3.1-YAP对细胞凋亡的影响.结果 转染si-CDK7后,HEC-1-A细胞中CDK7 mRNA和蛋白表达水平均显著下降(P<0.01),细胞活性显著降低(P<0.01),细胞凋亡率显著升高(P<0.01),YAP和磷酸化YAP蛋白及其下游蛋白Cyr16、CTGF表达水平明显下降(P<0.01);共转染pcDNA3.1-YAP可逆转沉默CDK7导致的细胞凋亡(P<0.01).结论 沉默CDK7可诱导YAP蛋白表达下调,促进子宫内膜癌HEC-1-A细胞凋亡.
Objective To investigate the effect of cyclin-dependent kinase 7(CDK7)on the apoptosis of endometrial cancer HEC-1-A cell by regulating of Yes-associated protein(YAP)expression.Methods The CDK7 expression was si-lenced in HEC-1-A cells with the siRNA interference technique.Then,the mRNA and protein expression levels of CDK7 were detected by qPCR and Western blotting respectively.The viability and apoptosis of HEC-1-A cells were detected by CCK-8 and flow cytometry after silencing CDK7.Expression levels of YAP,p-YAP protein and its downstream proteins Cyr16 and CTGF were detected by Western blotting after silencing CDK7.The effect of co-transfection with si-CDK7 and pcDNA3.1-YAP on the apoptosis of HEC-1-A cells was detected by apoptosis assay.Results After transfection with si-CDK7,the mRNA and protein expression levels of CDK7 in HEC-1-A cells were decreased significantly(P<0.01),the activity of HEC-1-A cells was significantly decreased(P<0.01),the apoptosis rate was significantly increased(P<0.01),and the expression levels of YAP,pYAP protein and its downstream proteins Cyr16 and CTGF were significantly decreased(P<0.01).Co-transfection of pcDNA3.1-YAP reversed the apoptosis caused by silenced CDK7(P<0.01).Conclusion Silencing CDK7 promoted the apoptosis of endometrial cancer HEC-1-A cells by downregulating the expression of YAP.
Objective To investigate the effect of cyclin-dependent kinase 7(CDK7)on the apoptosis of endometrial cancer HEC-1-A cell by regulating of Yes-associated protein(YAP)expression.Methods The CDK7 expression was si-lenced in HEC-1-A cells with the siRNA interference technique.Then,the mRNA and protein expression levels of CDK7 were detected by qPCR and Western blotting respectively.The viability and apoptosis of HEC-1-A cells were detected by CCK-8 and flow cytometry after silencing CDK7.Expression levels of YAP,p-YAP protein and its downstream proteins Cyr16 and CTGF were detected by Western blotting after silencing CDK7.The effect of co-transfection with si-CDK7 and pcDNA3.1-YAP on the apoptosis of HEC-1-A cells was detected by apoptosis assay.Results After transfection with si-CDK7,the mRNA and protein expression levels of CDK7 in HEC-1-A cells were decreased significantly(P<0.01),the activity of HEC-1-A cells was significantly decreased(P<0.01),the apoptosis rate was significantly increased(P<0.01),and the expression levels of YAP,pYAP protein and its downstream proteins Cyr16 and CTGF were significantly decreased(P<0.01).Co-transfection of pcDNA3.1-YAP reversed the apoptosis caused by silenced CDK7(P<0.01).Conclusion Silencing CDK7 promoted the apoptosis of endometrial cancer HEC-1-A cells by downregulating the expression of YAP.