SPOP对卵巢癌细胞生物学行为的影响及其分子机制的研究  

Effect of SPOP on Biological Behavior of Ovarian Cancer and Its Molecular Mechanism

作　　者：李若涵 袁冬 余秋波[2] 罗婧[1] 李颜希 孙一瑄 杨竹[1] Li Ruohan;Yuan Dong;Yu Qiubo;Luo Jing;Li Yanxi;Sun Yixuan;Yang Zhu(Second Affiliated Hospital of Chongqing Medical University,Chongqing,400016;Institute of Life Sciences,Chongqing Medical University,Chongqing,400016)

机构地区：[1]重庆医科大学附属第二医院,重庆400016 [2]重庆医科大学生命科学研究院,重庆400016

出　　处：《基因组学与应用生物学》2020年第4期1878-1885,共8页Genomics and Applied Biology

基　　金：重庆市卫生局医学科研计划项目(编号:2012-1-039);重庆市渝中区科技计划项目(编号:20110303)共同资助。

摘　　要：探讨人卵巢癌组织中斑点状蛋白(SPOP)的表达及对其卵巢癌细胞生物行为学的影响及相关机制。采用免疫组化技术分别检测正常卵巢组织10例、卵巢癌组织90例的组织芯片中SPOP的表达;体外培养永生化人卵巢癌细胞株OVCAR-3、SKOV3及永生化的人卵巢表皮上皮细胞株HOSE,RT-PCR、WB检测3种细胞株SPOP的表达。用慢病毒稳定转染OVCAR-3、HOSE细胞株,构建过表达、敲低SPOP的细胞株。流式细胞仪检测转染后细胞株的凋亡,CCK8检测转染后细胞株的增殖情况,Transwell小室检测转染后细胞株的迁移侵袭情况。WB检测转染后细胞株PI3K-Akt通路相关蛋白的表达情况。组织学免疫组化评分显示卵巢癌组织染色明显高于正常卵巢组织。细胞学实验显示,成功构建过表达、敲低SPOP的OVCAR-3细胞,过表达组、过表达空载组、敲低组、敲低空载组细胞凋亡及增殖能力无明显差异;各组细胞迁移及侵袭结果与空载组有明显差异,且差异有统计学意义(p<0.05)。过表达组细胞P-GSK3β(Ser9)表达水平明显低于敲低组(p<0.05),MMP-9表达水平明显高于敲低组(p<0.05)。SPOP在卵巢癌组织中表达上升,且具有促进卵巢癌细胞迁移侵袭的作用,其调节机制可能与p-GSK3β(Ser9)通路相关。To investigate the expression of SPOP in human ovarian cancer and its biological behavior and its related mechanism,Immunohistochemistry was used to detect the expression of SPOP in tissue microarray of 10 cases of normal ovarian tissue and 90 cases of ovarian cancer.Immortalized human ovarian cancer cell lines OVCAR-3,SKOV3 and immortalized human ovarian epithelial cell line HOSE were cultured in vitro and the expression of SPOP of the three cell lines were tested by WB and RT-PCR.Stable transfection of OVCAR-3 and HOSE cell lines with lentivirus and construction of over-expressed and knock-down SPOP cell lines.Flow cytometry,cck8,and Transwell chambers were used to detect apoptosis,proliferation,migration and invasion of Stable transfected cell lines.The expression of PI3K-Akt pathway-related proteins were detected by WB.Immunohistochemistry showed that ovarian cancer tissue staining was significantly higher than normal ovarian tissue.Cytology experiments showed that overexpression and knocked down SPOP OVCAR-3 cells were successfully constructed.There was no significant difference in apoptosis and proliferation between groups.The results of cell migration and invasion in each group were significantly different from those in no-load groups(p<0.05).The expression level of P-GSK3β(Ser9)in overexpression group was significantly lower than that of knockdown group(p<0.05),and the expression of MMP-9 in overexpression group was significantly higher than that of knockdown group(p<0.05).The expression of the SPOP in ovarian cancer is increased,and it has the role of promoting migration and invasion of ovarian cancer cells.The underlying mechanism was related to the down-regulation of p-GSK3β(Ser9)by SPOP and the up-regulation of MMP-9.

关 键 词：人卵巢癌 卵巢癌细胞 SPOP 迁移 侵袭 MMP9 

分 类 号：R737.31[医药卫生—肿瘤]