微小RNA-9-5p靶向HIC1降低乳腺癌细胞对多柔比星敏感性的研究  

Study on microRNA-9-5p reducing sensitivity of breast cancer cells to doxorubicin through targeting HIC1

作　　者：高航 赵峰[1] 吴衍[1] 裴文江 钟明 顾岩[1] 郭善禹[1] 戴谦诚[1] 张伟[1] GAO Hang;ZHAFeng;WU Yan;PEI Wenjiang;ZHONG Ming;GU Yan;GUO Shanyu;DAI Qiancheng;ZHANG Wei(Department of General Surgery,The Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院普外科,上海200011

出　　处：《外科理论与实践》2020年第3期227-233,共7页Journal of Surgery Concepts & Practice

基　　金：国家自然科学基金(81572760)。

摘　　要：目的:探讨微小RNA-9-5p(mi R-9-5p)调控肿瘤高甲基化基因1(hypermethylated in cancer 1,HIC1)对乳腺癌MDA-MB-231细胞对化疗药多柔比星敏感性的影响及其作用机制。方法:(1)使用慢病毒转染建立上调或下调mi R-9-5p细胞,加入不同浓度多柔比星后,用CCK-8和流式细胞仪检测细胞活力和凋亡情况。(2)通过反转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和蛋白质印迹试验检测上调或下调miR-9-5p时HIC1的表达情况。通过数据库资料、双荧光素酶报告基因系统验证miR-9-5p与HIC1的关系。(3)下调HIC1,观察加入多柔比星后细胞活力和凋亡情况。结果:(1)上调miR-9-5p可提高细胞活力、抑制凋亡。下调mi R-9-5p可降低细胞活力、促进凋亡。(2)miR-9-5p靶向HIC1,两者的表达呈负相关。(3)抑制HIC1的表达,可逆转miR-9-5p下调的作用。结论:miR-9-5p可通过下调HIC1降低乳腺癌细胞对多柔比星的敏感性。Objective To study the effect of microRNA-9-5 p(miR-9-5 p)which regulate hypermethylated in cancer 1(HIC1)in reducing sensitivity of breast cancer MDA-MB-231 cells to doxorubicin(DOX)and the mechanisms.Methods①Up-regulation or down-regulation of miR-9-5 p in cells was constructed by lentivirus transfection.Cell viability and apoptosis were detected with CCK-8 and flow cytometry after adding different concentrations of DOC.②Expression of HIC1 was detected by reverse transcription-polymerase chain reaction and Western blotting when miRNA-9-5 p was overexpressed or inhibited.The interaction between miR-9-5 p and HIC1 was analyzed by database and luciferase assay.③HIC1 was down-regulated to detect cell viability and apoptosis after adding different concentrations of DOX.Results①Up-regulation of miR-9-5 p increased cell viability,and reduced apoptosis.Down-regulation of miR-9-5 p reduced cell viability and increased apoptosis.②miR-9-5 p directly targeted HIC1 and negative relationship of expression was present between miR-9-5 p and HIC1.③Down-regulation of HIC1 could reverse the down-regulation effect of miR-9-5 p.Conclusions miR-9-5 p reduces the sensitivity of breast cancer cells to DOX through down-regulation of HIC1.

关 键 词：乳腺癌 微小RNA-9-5p 肿瘤高甲基化基因1 多柔比星 敏感性 

分 类 号：R737.9[医药卫生—肿瘤]