N~6-甲基腺苷甲基化调节因子RNA结合基序蛋白15在高级别浆液性卵巢癌中的生物学功能研究  

Biological function of N6-methyladenosine methylation regulator RNA-binding motif protein 15 in high-grade serous ovarian cancer

作　　者：周芝润 龚静亚[2] 朱维培[2] Zhou Zhirun;Gong Jingya;Zhu Weipei(Department of Gynecology,the Second Affiliated Hospital of Soochow University,the First People’s Hospital of Taicang,Taicang 215400,China;Department of Obstetrics and Gynecology,the Second Affiliated Hospital of Soochow University,Taicang 215400,China)

机构地区：[1]苏州大学附属第二医院妇科妇瘤组太仓市第一人民医院,太仓215400 [2]苏州大学附属第二医院妇产科,太仓215400

出　　处：《中华实验外科杂志》2023年第8期1581-1584,共4页Chinese Journal of Experimental Surgery

基　　金：助医行动-肿瘤学科研能力建设项目。

摘　　要：目的探讨RNA结合基序蛋白15(RBM15)与高级别浆液性卵巢癌(HGSOC)患者预后的关系及对肿瘤细胞的生物学功能的影响。方法在基因表达数据库(GEO)中用GSE69428和GSE40595数据集分析RBM15在HGSOC及正常组织中的表达, 在Kaplan-Meier Plotter在线网站分析RBM15与HGSOC预后的相关性。通过实时荧光定量聚合酶链反应(RT-qPCR)方法检测人正常卵巢上皮细胞以及卵巢癌细胞系中RBM15的表达。运用慢病毒转导技术进行RBM15敲降。采用平板克隆实验、细胞计数试剂盒(CCK-8)法细胞增殖实验Transwell迁移侵袭实验、药物毒性实验检测各组细胞的集落形成、增殖、迁移、侵袭能力及顺铂敏感性。采用t检验或单因素方差分析进行统计分析。结果在GSE69428和GSE40595数据集中, RBM15基因在肿瘤组织中显著高表达(7.53±0.44比6.40±0.37, t=6.215, P<0.01;6.18±0.46比5.61±0.20, t=4.487, P<0.01), 且高表达组无进展生存期及总生存期显著高于低表达组。与正常人上皮卵巢细胞系IOSE80比较, RBM15在卵巢癌细胞系SKOV3及OVCAR3中的表达显著上调(1.56±0.09比1.00±0.04, t=10.190, P<0.01;2.19±0.03比1.00±0.04, t=42.360, P<0.01)。shRBM15组细胞表达明显低于shNC组细胞(SKOV3:0.59±0.05、0.78±0.04比1.00±0.04, F=62.991, P<0.01;OVCAR3:0.27±0.01、0.45±0.03比1.01±0.13, F=66.290, P<0.01)。shRBM15组细胞的集落形成能力明显高于shNC组(SKOV3:307±40、287±13比209±26, F=9.715, P<0.05;OVCAR3:299±37、192±24比121±12, F=34.591, P<0.05)。shRBM15组细胞的体外增殖能力明显高于shNC组(SKOV3:72 h 1.43±0.03、1.52±0.09比1.12±0.04, F=40.240, P<0.01;OVCAR3:72 h 2.52±0.03、2.59±0.09比2.36±0.05, F=9.753, P<0.05)。shRBM15组细胞的迁移能力明显高于shNC组(SKOV3:656±87、655±99比395±95, F=12.771, P<0.01;OVCAR3:240±9、237±19比185±24, F=13.729, P<0.01)。shRBM15组细胞的侵袭能力明显高于shNC组(SKOV3:971±109、876±160比649±65, F=9.803, P<0.01;OVCAR3:325±37、349�Objective To investigate the relationship between RNA-binding motif protein 15(RBM15)and the prognosis of patients with high-grade serous ovarian cancer(HGSOC)and the effect on the biological function of cancer cells.Methods The expression of RBM15 in HGSOC and normal tissues was analyzed in two datasets,GSE69428 and GSE40595,in the Gene Expression Database(GEO),and the correlation between RBM15 and the prognosis of HGSOC was analyzed in the Kaplan-Meier Plotter online website.RT-qPCR was used to detect the expression level of RBM15 in human normal ovarian epithelial cells as well as ovarian cancer cell lines.The lentiviral shRNA was used to infiltrate ovarian cancer cells to construct a stable transient cell line with RBM15 knockdown,and the interference efficiency was detected by Western blotting as well as RT-qPCR.The proliferation of cell clones was detected by plate clone formation assay and cell counting kit-8(CCK-8)assay,the migration and invasion ability of cells was detected by Transwell assay,and the resistance of cisplatin was detected by drug toxicity assay.Statistical analysis was performed using t-test or one-way ANOVA.Results In the GSE69428 and GSE40595 datasets,the RBM15 gene was significantly highly expressed in tumor tissues(7.53±0.44 vs.6.40±0.37,t=6.215,P<0.01;6.18±0.46 vs.5.61±0.20,t=4.487,P<0.01),and the progression-free survival and overall survival in the high-expression group were significantly higher than those in the low expression group.As compared with the normal human epithelial ovarian cell line IOSE80,the expression of RBM15 was significantly up-regulated in the ovarian cancer cell lines SKOV3 and OVCAR3(1.56±0.09 vs.1.00±0.04,t=10.190,P<0.01;2.19±0.03 vs.1.00±0.04,t=42.360,P<0.01).The expression of cells in the shRBM15 group was significantly lower than that the shNC group(SKOV3:0.59±0.05,0.78±0.04 vs.1.00±0.04,F=62.991,P<0.01;OVCAR3:0.27±0.01,0.45±0.03 vs.1.01±0.13,F=66.290,P<0.01).The colony formation ability of cells in the shRBM15 group was significantly higher

关 键 词：高级别浆液性卵巢癌 RNA结合基序蛋白15 生物学功能 预后 

分 类 号：R737.31[医药卫生—肿瘤]