CASC5通过BUB1促进乳腺癌发生发展的分子机制研究  

Molecular mechanism of CASC5 promoting breast cancer development through BUB1

作　　者：何贵省 陈恒余 吕小玲 宋韬 陈秀秀 符广华 吴煌福 陈运景 苏亚静 郑武平[1] HE Gui-sheng;CHEN Heng-yu;LYU Xiao-ling;SONG Tao;CHEN Xiu-xiu;FU Guang-hua;WU Huang-fu;CHEN Yun-jing;SU Ya-jing;ZHENG Wu-ping(Department of Breast and Thyroid Surgery,Second Affiliated Hospital of Hainan Medical College,Haikou Hainan 570100,China;Department of Otolaryngology,Second Affiliated Hospital of Hainan Medical College,Haikou Hainan 570100,China;Department of General Surgery,928th Hospital of Joint Logistics Support Force of PLA,Haikou Hainan 571199,China)

机构地区：[1]海南医学院第二附属医院乳甲外科,海南海口570100 [2]海南医学院第二附属医院耳鼻喉科,海南海口570100 [3]中国人民解放军联勤保障部队第九二八医院普外科,海南海口571199

出　　处：《局解手术学杂志》2023年第4期297-303,共7页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然科学基金(82203813);海南省卫生健康委员会基金项目(21A200211);2021年海南省重点研发计划项目(ZDYF2021SHFZ248)。

摘　　要：目的探究易感性癌症候选基因5(CASC5)对乳腺癌发生发展的作用及其相关机制。方法收集50例乳腺癌患者的肿瘤组织及其癌旁组织,采用qRT-PCR检测CASC5的表达水平,并分析其与患者临床病理特征的关系。通过脂质体转染法将si-CASC5、si-BUB1和si-NC转染至MDA-MB-231和MCF-7细胞,采用qRT-PCR和Western blot检测细胞CASC5、BUB1表达水平,CCK-8检测细胞增殖能力,Transwell检测细胞侵袭能力。利用String数据库分析CASC5相关蛋白,并在MCF7细胞中验证。通过免疫共沉淀检测CASC5与BUB1的结合情况,qRT-PCR验证BUB1在肿瘤组织和癌旁组织中的表达水平,及其与CASC5表达水平的相关性。结果与癌旁组织相比,肿瘤组织中CASC5表达水平升高(P<0.05),CASC5表达水平与患者TNM分期、淋巴结转移相关(P<0.05)。与si-NC组相比,si-CASC5组MDA-MB-231和MCF-7细胞中CASC5表达水平降低(P<0.05),细胞增殖能力及侵袭能力减弱(P<0.05)。生物信息学预测CASC5可能与BUB1、NSL1、NDC80和MIS12等相互作用。与si-NC组相比,si-CASC5组MDA-MB-231细胞中BUB1表达水平降低(P<0.05),NSL1、NDC80和MIS12表达水平无统计学差异(P>0.05)。CASC5免疫沉淀产物中可检测到BUB1蛋白,BUB1免疫沉淀产物中可检测到CASC5蛋白。与si-NC组相比,si-BUB1组MDA-MB-231细胞中BUB1表达水平降低(P<0.05),细胞增殖能力及侵袭能力减弱(P<0.05)。与癌旁组织相比,肿瘤组织中BUB1表达水平升高(P<0.05),CASC5表达水平与BUB1表达水平呈正相关(r=0.4815)。结论CASC5可通过与BUB1相互作用,调控BUB1蛋白表达从而促进乳腺癌发生发展。Objective To investigate the role of cancer susceptibility candidate 5(CASC5)in the development of breast cancer and its related mechanisms.Methods Tumor tissues and paracancerous tissues from 50 breast cancer patients were collected,and the expression level of CASC5 was detected by qRT-PCR and its relationship with clinicopathological features was analyzed.si-CASC5,si-BUB1 and si-NC were transfected into MDA-MB-231 and MCF-7 cells by liposome transfection.The expression levels of cellular CASC5 and BUB1 were detected by qRT-PCR and Western blot,the proliferation ability of cells was detected by CCK-8 and the invasion ability of cells was detected by Transwell.CASC5-related proteins were analyzed by the String database and verified in MCF7 cells.Binding of CASC5 to BUB1 was detected by immunoprecipitation.qRT-PCR was used to verify the expression levels of BUB1 in tumor tissues and paracancerous tissues,and its correlation with the expression level of CASC5.Results Compared with paracancerous tissues,the expression level of CASC5 in tumor tissues was increased(P<0.05),and the expression level of CASC5 was correlated with TNM stage and lymph node metastasis(P<0.05).Compared with si-NC group,the expression level of CASC5 in MDA-MB-231 and MCF-7 cells of si-CASC5 group was decreased(P<0.05),the cell proliferation ability and the cell invasion ability were both decreased(P<0.05).Bioinformatics predicted that CASC5 may interact with BUB1,NSL1,NDC80 and MIS12.Compared with si-NC group,the expression level of BUB1 in MDA-MB-231 cells of si-CASC5 group was decreased(P<0.05),and the expression levels of NSL1,NDC80 and MIS12 were not significantly different(P>0.05).BUB1 protein was detected in the immunoprecipitation product of CASC5,and CASC5 protein was detected in the immunoprecipitation product of BUB1.Com-pared with si-NC group,the expression level of BUB1 in MDA-MB-231 cells of si-BUB1 group was decreased(P<0.05),the cell proliferation abil-doiity and the cell invasion ability were both decreased(P<0.05).The expressi

关 键 词：乳腺癌 易感性癌症候选基因5 BUB1 免疫共沉淀 

分 类 号：R737.9[医药卫生—肿瘤]