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

doi:10.16139/j.1007-9610.2020.03.011

摘要/Abstract

摘要：

目的:探讨微小RNA-9-5p（miR-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miR-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miR-9-5p可降低细胞活力、促进凋亡。②miR-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。

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

Abstract:

Objective To study the effect of microRNA-9-5p (miR-9-5p) which regulate hypermethylated in cancer 1 (HIC1) in reducing sensitivity of breast cancer MDA-MB-231 cells to doxorubicin (DOX) and the mechanisms. Methods ①Up-regulation or down-regulation of miR-9-5p 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-9-5p was overexpressed or inhibited. The interaction between miR-9-5p 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. Results ①Up-regulation of miR-9-5p increased cell viability, and reduced apoptosis. Down-regulation of miR-9-5p reduced cell viability and increased apoptosis. ②miR-9-5p directly targeted HIC1 and negative relationship of expression was present between miR-9-5p and HIC1. ③Down-regulation of HIC1 could reverse the down-regulation effect of miR-9-5p. Conclusions miR-9-5p reduces the sensitivity of breast cancer cells to DOX through down-regulation of HIC1.

Key words: Breast cancer, MicroRNA-9-5p, Hypermethylated in cancer 1, Doxorubicin, Sensitivity

中图分类号:

R737.9

高航, 赵峰, 吴衍, 裴文江, 钟明, 顾岩, 郭善禹, 戴谦诚, 张伟. 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[J]. 外科理论与实践, 2020, 25(03): 227-233.

GAO Hang, ZHAO Feng, WU Yan, PEI Wenjiang, ZHONG Ming, GU Yan, GUO Shanyu, DAI Qiancheng, ZHANG Wei. Study on microRNA-9-5p reducing sensitivity of breast cancer cells to doxorubicin through targeting HIC1[J]. Journal of Surgery Concepts & Practice, 2020, 25(03): 227-233.

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