circ_0000515/miR-1296-5p/CDK2轴在乳腺癌中的作用  

The role of circ_0000515/miR-1296-5p/CDK2 axis in breast cancer

作　　者：杨琴[1] 王超[2] 蔺博涵 虞绮雯 李涛[2] 孙晓春[2] YANG Qin;WANG Chao;LIN Bohan;YU Qiwen;LI Tao;SUN Xiaochun(Women′s Health Department,the Fourth Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212002;School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学附属四院妇女保健科,江苏镇江212002 [2]江苏大学医学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2023年第4期317-322,共6页Journal of Jiangsu University：Medicine Edition

基　　金：镇江市社会发展项目(SH2021029)。

摘　　要：目的:探究circ_0000515在乳腺癌进展中的可能作用机制。方法:通过GEO数据库下载乳腺癌circRNA芯片数据集(GSE101123)并采用R语言进行表达差异分析,筛选出目标circRNA分子作为研究对象并检测其在乳腺癌细胞株和组织中的表达;生物信息学分析与选择该circRNA的目的miRNA,及后者对应的靶向mRNA,并采用双荧光素酶实验和蛋白质印迹进行验证;将目标circRNA的siRNA转染人乳腺癌MCF-7和MDA-MB-231细胞,CCK-8实验和划痕实验观察细胞的增殖和迁移,荧光定量PCR(qRT-PCR)检测RNA表达,蛋白质印迹实验检测mRNA编码蛋白的表达。结果:选择和下载GSE101123数据集后用R语言筛选出5个高表达的circRNA分子,选定其中的circ_0000515作为研究对象,生物信息学分析得出circ_0000515、miR-1296-5p和细胞周期蛋白依赖性激酶2(cyclin-dependent kinase 2,CDK2)三者之间可能存在靶向关系。qRT-PCR结果显示,circ_0000515中乳腺癌细胞株MCF-7和MDA-MB-231中的表达明显高于人乳腺上皮细胞株MCF-10A;在乳腺癌组织中的表达明显高于癌旁组织中的表达。抑制circ_0000515表达后,乳腺癌细胞增殖和迁移能力较对照组显著下降,miR-1296-5p表达明显增高;双荧光素酶实验证实circ_0000515与miR-1296-5p之间存在靶向关系。蛋白质印迹实验证实抑制circ_0000515表达后CDK2蛋白表达明显减低。结论:circ_0000515在乳腺癌中高表达,可能通过circ_0000515/miR-1296-5p/CDK2作用轴影响乳腺癌进展。Objective:To explore the role and underlying mechanism of circ_0000515 in the progression of breast cancer.Methods:The circRNAs chip dataset(GSE101123)of breast cancer was downloaded from GEO database and analyzed by R language.The target circRNA was selected and its expression in breast cancer cell lines and tissues were detected.The target miRNA of the circRNA and the target mRNA of the miRNA were selected by bioinformatics analysis,and verified by double luciferase experiment and Western blotting.The siRNA of circRNA was transfected into MCF-7 and MDA-MB-231.The cell proliferation and migration were measured by CCK-8 and scratch assay.The expression of RNA were detected by qRT-PCR.The expression of mRNAs-encoded protein was detected by Western blotting.Results:GSE101123 dataset was selected and downloaded.Five highly expressed circRNAs molecules were screened by R language,and circ_0000515 was selected as the research object for subsequent study.Bioinformatics analysis showed that there may be a targeted relationship among circ_0000515,miR-1296-5p and cell cycle-dependent kinase 2(CDK2).qRT-PCR results showed that the expression of circ_0000515 in breast cancer cell lines was higher than that in normal cell lines,and its expression in cancer tissues was higher than that in paracancerous tissues.After knockdown the expression of circ_0000515,the proliferation and migration ability of breast cancer cells decreased significantly,while the expression of miR-1296-5p increased.Double luciferase assay confirmed that there was a targeted relationship between circ_0000515 and miR-1296-5p.Western blotting showed that the expression of CDK2 decreased after knocking down the expression of circ_0000515.Conclusion:circ_0000515 is highly expressed in breast cancer.circ_0000515/miR-1296-5p/CDK2 axis may play a role in the progression of breast cancer.

关 键 词：circ_0000515 乳腺癌 细胞周期蛋白依赖性激酶2 miR-1296-5p 机制 

分 类 号：R737.9[医药卫生—肿瘤]