摘要
目的:探讨细胞骨架调节剂(lncRNA cytoskeleton regulator,CYTOR)是否通过靶向调控miR-503来促进细胞周期蛋白E1(cyclin E1,CCNE1)影响上皮性卵巢癌增殖。方法:实时定量聚合酶链反应(RT-qPCR)检测CYTOR在不同临床分期、病理分级和有无淋巴结转移患者人上皮性卵巢癌组织、癌旁组织和正常组织以及不同卵巢癌细胞中的表达;starBase数据库(网址:http://starbase.sysu.edu.cn/)预测分析CYTOR和miR-503以及miR-503与CCNE1之间的关系,同时荧光素酶实验进行验证;RT-qPCR和免疫荧光分别检测miR-503与CCNE1在人上皮性卵巢癌组织、癌旁组织和正常组织以及不同卵巢癌细胞中的表达;RNA转染技术在A2780中细胞分别沉默CYTOR、过表达miR-503和沉默CCNE1表达,CCK-8检测各组细胞增殖能力变化;RT-qPCR和免疫荧光检测沉默CYTOR后miR-503和CCNE1的表达以及过表达miR-503后CCNE1的表达。结果:CYTOR和CCNE1在上皮性卵巢癌组织中表达增加,同时在临床分期III-IV期、病理分级G_(3)级、淋巴结转移阳性患者肿瘤组织中表达同样显著增加且导致卵巢癌患者术后预后较差;通过数据库分析CYTOR可直接靶向抑制miR-503,而miR-503可直接靶向抑制CCNE1表达,双荧光素酶实验结果与之相同;CYTOR和CCNE1促进上皮性卵巢癌增殖,而miR-503抑制上皮性卵巢癌增殖作用。结论:lncRNA CYTOR通过调控miR-503/CCNE1轴促进上皮性卵巢癌的增殖。
Objective:To explore whether lncRNA CYTOR promotes cell proliferation via miR-503/CCNE1 axis in epithelial ovarian cancer.Methods:lncRNA CYTOR mRNA levels were measured by RT-qPCR in human epithelial ovarian cancer tissues with pathological grade,lymph node metasatsis,and clinical stage.lncRNA CYTOR expression in both normal tissues and carcinoma tissues and in ovarian cancer cell lines was examined by RT-qPCR.Luciferase reporter assay the target sites were analyzed using StarBase online software version 3.0(between CYTOR and miR-503,miR-503 and CCNE1).The expression levels of miR-503 and CCNE1 in normal tissues,carcinoma tissues and ovarian cancer cell lines were detected by RT-qPCR and immunofluorescence analysis.Overexpression of miR-503 and silencing of CYTOR and CCNE1 in A2780 cells were achieved by transfection with either miR-503 mimics or CYTOR,CCNE1 inhibitor.CCK-8 assay was used to detect the cell proliferative ability.The expression of miR-503 and CCNE1 after silencing CYTOR and the expression of CCNE1 after overexpression of miR-503 were detected by qRT-PCR and immunofluorescence analysis.Results:CYTOR expression was elevated both in ovarian cancer tissues and cells.CYTOR and CCNE1 expression was significantly up-regulated in ovarian cancer tissues with pathological grade G_(3),positive lymph node metasatsis,and clinical stage and associated with poor prognosis.The results of dual luciferase expermient wer the same CYTOR can directly target and inhibit miR-503,while miR-503 can directly target and inhibit the expression of CCNE1 through database analysis.CYTOR and CCNE1 strongly promoted ovarian cancer proliferation,whereas miR-503 exerted the opposite effect.Conclusion:lncRNA CYTOR might promote the proliferation of epithelial ovarian cancer by regulating the miR-503/CCNE1 axis.
作者
郑轶
周玉
刘淑玉
何玉
ZHENG Yi;ZHOU Yu;LIU Shuyu;HE Yu(Department of Obstetrics and Gynecology,the First Affiliated Hospital of Bengbu Medical College,Anhui Bengbu 230000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2022年第22期4155-4161,共7页
Journal of Modern Oncology
基金
蚌埠医学院自然科学基金(编号:2020byzd149)。
