摘要
为了探究AKAP12对卵巢癌(ovarian cancer,OV)细胞恶性行为的特异性作用,并基于枢纽基因(hub-Genes)构建OV预后风险预测模型,应用生物信息学技术,将“ovarian cancer”作为关键词,从GeneCards数据库得到OV相关基因,通过基因表达综合数据库(GEO)筛选具有显著差异的OV相关枢纽基因并进行功能分析.通过单因素Cox回归分析筛选出与OV样本总生存期显著相关的hub-Genes;用LASSO算法构建卵巢癌预后风险预测模型,并通过Kaplan-Meier(KM)生存曲线、单因素多因素Cox回归分析以及ROC曲线检测该模型对OV患者的预后预测能力,通过SangerBox在线分析AKAP12与OV患者预后的相关性.细胞转染实现AKAP12过表达后,通过MTT实验检测细胞增殖能力,流式细胞术检测细胞凋亡能力,Transwell检测细胞侵袭能力,Western blot检测AMPK信号通路相关蛋白的表达.结果显示:①从GeneCards数据库中获得8378个OV相关基因,其中82个是在正常组织与OV组织中存在显著差异的hub-Genes;②成功构建了基于5个hub-Genes(BIRC5、EPCAM、AKAP12、PDGFRA、CLDN4)组成的OV风险预测模型,该模型能较好地预测OV患者的预后情况;③SangerBox在线分析结果表明,AKAP12高表达患者的预后更差;④细胞实验证明,AKAP12过表达可明显促进细胞的增殖和侵袭,抑制细胞的凋亡;⑤过表达AKAP12可显著降低AMPK磷酸化水平,明显提高mTOR磷酸化水平.总之,本研究基于5个hub-Genes构建的特征风险模型能有效预测OV患者的预后情况,AKAP12可能通过AMPK/mTOR信号通路诱导OV细胞的增殖和侵袭,抑制OV细胞凋亡.
In order to explore the specific effect of AKAP12 on malignant behavior of ovarian cancer(OV)cells and construct a prognostic risk prediction model of OV based on hub-Genes,bioinformatics technology was employed,and ovarian cancer was used as a keyword to obtain OV-associated genes from the GeneCards database.OV-associated hub-Genes with significant differ-ences were screened through the comprehensive gene expression database(GEO)and their functions were analyzed.The hub-Genes which were significantly related to the overall survival of OV samples were screened by univariate Cox regression analyRole and mechanism of AKAP12 regulation on AMPK/mTOR signaling pathway mediating cell proliferation,apoptosis and invasion in ovarian cancer cells WANG Yinghong1,YUAN Min2,TIE Hailong3,LIU Xiaoshan2,ZHU Changjun4,XIONG Tingchuan2(1.Institute of Medical Sciences of Xinjiang Medical University,Center of Heath Management,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;2.Department of Gynecologic Surgery,The 3rd Affiliated Teaching Hospital of Xinjiang Medical University(Affiliated Cancer Hospital),Urumqi 830011,China;3.Department of Science and Technology Management,The 3rd Affiliated Teaching Hospital of Xinjiang Medical University(Affiliated Cancer Hospital),Urumqi 830011,China;4.Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin 300387,China)Abstract:In order to explore the specific effect of AKAP12 on malignant behavior of ovarian cancer(OV)cells and construct a prognostic risk prediction model of OV based on hub-Genes,bioinformatics technology was employed,and ovarian cancer was used as a keyword to obtain OV-associated genes from the GeneCards database.OV-associated hub-Genes with significant differ-ences were screened through the comprehensive gene expression database(GEO)and their functions were analyzed.The hub-Genes which were significantly related to the overall survival of OV samples were screened by univariate Cox regression analy-sis.LASSO algorithm was used to construct a prognostic risk prediction model for ovarian cancer.Kaplan-Meier(KM)survival curve,univariate and multivariate Cox regression analysis and ROC curve were used to test the predictive ability of this model for OV patients,and SangerBox was used to analyze the correlation between AKAP12 and OV patients′prognosis online.After AKAP12 overexpression was achieved,MTT assay was used to detect cell proliferation,flow cytometry was used to detect cell apoptosis,transwell was used to detect cell invasion,and Western blot was used to detect the expression of AMPK signaling pathway-related proteins.The results were as follows:①8378 OV-related genes were obtained from GeneCards data base,among which 82 OV-related hub-Genes had significant differences between normal and OV tissues;②An OV risk prediction model based on five hub-Genes(BIRC5,EP-CAM,AKAP12,PDGFRA,CLDN4)was successfully constructed,which could predict the prognosis of patients with OV;③SangerBox online analysis showed that patients with high AKAP12 expres-sion had worse prognosis;④The overexpression of AKAP12 can significantly promote cell proliferation and invasion,and in-hibit cell apoptosis;⑤Overexpression of AKAP12 significantly decreased AMPK phosphorylation and significantly increased mTOR phosphorylation.In conclusion,in this study,the characteristic risk model constructed based on five hub-Genes can effectively predict the prognosis of OV patients.AKAP12 can induce the proliferation and invasion of OV cells through AMPK/mTOR signaling pathway,and inhibit the apoptosis of OV cells.
作者
王迎洪
袁敏
铁海龙
刘小山
朱长军
熊廷川
WANG Yinghong;YUAN Min;TIE Hailong;LIU Xiaoshan;ZHU Changjun;XIONG Tingchuan(Institute of Medical Sciences of Xinjiang Medical University,Center of Heath Management,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;Department of Gynecologic Surgery,The 3rd Affiliated Teaching Hospital of Xinjiang Medical University(Affiliated Cancer Hospital),Urumqi 830011,China;Department of Science and Technology Management,The 3rd Affiliated Teaching Hospital of Xinjiang Medical University(Affiliated Cancer Hospital),Urumqi 830011,China;Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin 300387,China)
出处
《天津师范大学学报(自然科学版)》
CAS
北大核心
2024年第6期13-19,共7页
Journal of Tianjin Normal University:Natural Science Edition
基金
新疆医科大学医学科学研究所开放课题资助项目(YXYJ20230206)
新疆维吾尔自治区自然科学基金资助项目(2023D01C128).