RAD51AP1通过CDK1/PI3K/AKT通路影响卵巢癌细胞增殖、细胞周期和凋亡  

RAD51AP1 affects the proliferation,cell cycle and apoptosis of ovarian cancer cell through CDK1/PI3K/AKT pathway

作　　者：林丽丽[1] 莫新宇[1] 刘莉[1] 莫芳 LIN Lili;MO Xinyu;LIU Li;MO Fang(Department of Obstetrics and Gynecology,Nanxishan Hospital of Guangxi Zhuang Autonomous Region,The Second People’s Hospital of Guangxi Zhuang Autonomous Region,Guilin,Guangxi 541002,China)

机构地区：[1]广西壮族自治区南溪山医院(广西壮族自治区第二人民医院)妇产科,广西桂林541002

出　　处：《中国优生与遗传杂志》2024年第9期1812-1819,共8页Chinese Journal of Birth Health & Heredity

基　　金：桂林市科学研究与技术开发计划项目(20210227-9-1)。

摘　　要：目的探讨RAD51相关蛋白1(RAD51AP1)通过调控细胞周期蛋白依赖性激酶1(CDK1)/磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)通路对卵巢癌细胞(OVCAR3)增殖、细胞周期和凋亡的影响及其分子机制。方法通过生信数据库筛选差异表达基因,及对RAD51AP1在卵巢癌患者中的表达和总生存期进行分析,qRT-PCR和Western blot检测卵巢癌细胞系中RAD51AP1表达;体外培养OVCAR3细胞,分为Ctrl组、NC组、RAD51AP1-KD组、RAD51AP1-KD+CDK1组,qRT-PCR和Westernblot检测RAD51AP1、CDK1和PI3K/AKT通路相关蛋白表达,CCK8和平板克隆形成实验检测细胞增殖,细胞周期和凋亡由流式细胞术测定。结果差异表达基因RAD51AP1在卵巢癌组织和细胞系中显著上调(P<0.05),且其高表达患者的总生存期时间短(P<0.05);敲低RAD51AP1显著降低了OVCAR3细胞活力和克隆形成能力,阻滞细胞在G0/G1期,促进凋亡,下调CDK1、PI3K和P-AKT的表达(P<0.05),过表达CDK1则部分逆转上述指标(P<0.05)。结论RAD51AP1在卵巢癌细胞中高表达,敲低RAD51AP1可通过调节CDK1/PI3K/AKT通路抑制卵巢癌细胞增殖,诱导细胞周期阻滞和凋亡。Objective To investigate the effect and molecular mechanism of RAD51-associated protein 1(RAD51AP1)on the proliferation,cell cycle and apoptosis of ovarian cancer(OC)cell(OVCAR3)by regulating cyclin dependent kinase 1(CDK1)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods The differentially expressed genes were screened by bioinformatics database,and the expression of RAD51AP1 and overall survival(OS)in OC patients were analyzed.The expression of RAD51AP1 in OC cell lines was detected by qRT-PCR and Western blot.OVCAR3 cells were treated according to Ctrl group,NC group,RAD51AP1-KD group and RAD51AP1-KD+CDK1 group.qRT-PCR and Western blot were used to detect the expressions of RAD51AP1,CDK1 and PI3K/AKT pathway related proteins.CCK8 and colony formation assay were used to assess cell proliferation,and flow cytometry was used to detect cell cycle and apoptosis.Results RAD51AP1 was significantly up-regulated in OC tissues and cell lines(P<0.05),and the OS time of patients with high RAD51AP1 expression was shorter(P<0.05).Knockdown of RAD51AP1 significantly down-regulated the expression of CDK1,PI3K and P-AKT,decreased the viability and colony formation ability of OVCAR3 cells,arrested the cells in G0/G1 phase,promoted apoptosis(P<0.05),while overexpression of CDK1 partially reversed the above indicators(P<0.05).Conclusion RAD51AP1 is highly expressed in OC cells.Knockdown RAD51AP1 could inhibit the proliferation,induce cell cycle arrest and apoptosis by regulating CDK1/PI3K/AKT pathway in OC cell.

关 键 词：RAD51相关蛋白1(AP1) 细胞周期蛋白依赖性激酶1(CDK1) 磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路 卵巢癌 细胞周期 

分 类 号：R737.31[医药卫生—肿瘤]