Mechanism of curcumin improving cisplatin resistance of human ovarian cancer cells based on the targeted regulation of flap-like endonuclidene 1 by microRNA-135b-5p
- Author:
- No author available
- Journal Title:
- China Medicine
- Issue:
- 12
- DOI:
- 10.3760/j.issn.1673-4777.2022.12.015
- Key Word:
- 卵巢癌;微小RNA-135b-5p;瓣状核酸内切酶1;姜黄素;顺铂耐药
Abstract： 目的 基于微小RNA-135b-5p(miR-135b-5p)靶向调控瓣状核酸内切酶1(FEN1)探究姜黄素对人卵巢癌细胞顺铂耐药的改善机制.方法 取对数生长期人卵巢癌顺铂耐药细胞株OVCAR-3/DDP,将细胞分为对照组、顺铂组、姜黄素组、顺铂+姜黄素组、顺铂+姜黄素+阴性对照抑制剂组、顺铂+姜黄素+miR-135b-5p抑制剂组、顺铂+姜黄素+miR-135b-5p模拟物组.对照组不予处理,其余各组分别予顺铂和/或姜黄素,后3组细胞采用脂质体转染法分别转染绿色荧光蛋白质粒(pGFP)-阴性对照抑制剂、pGFP-miR-135b-5p抑制剂、pGFP-miR-135b-5p模拟物载体.各组给药处理72 h后,采用实时荧光定量聚合酶链反应检测各组细胞miR-135b-5p及FEN1 mRNA表达量;蛋白质印迹法检测各组细胞FEN1蛋白表达;双荧光素酶靶标实验验证miR-135b-5p与FEN1之间的作用关系;噻唑盐比色法检测各组细胞增殖抑制率;流式细胞仪检测各组细胞凋亡率.结果 姜黄素组、顺铂+姜黄素组、顺铂+姜黄素+阴性对照抑制剂组、顺铂+姜黄素+miR-135b-5p抑制剂组、顺铂+姜黄素+miR-135b-5p模拟物组miR-135b-5p表达量、增殖抑制率及细胞凋亡率均高于对照组和顺铂组,FEN1 mRNA及蛋白表达量均低于对照组和顺铂组(均P<0.05).脂质体转染的3组中,顺铂+姜黄素+miR-135b-5p抑制剂组miR-135b-5p表达量、增殖抑制率及细胞凋亡率最低[(0.75±0.10)、(34.399±6.055)％、(17.3±3.1)％],FEN1 mRNA和蛋白表达量最高[(0.77±0.11)、(0.69±0.09)];顺铂+姜黄素+miR-135b-5p模拟物组miR-135b-5p表达量、增殖抑制率及细胞凋亡率最高[(1.68±0.26)、(80.365±10.617)％、(48.7±8.4)％],FEN1 mRNA和蛋白表达量最低[(0.33±0.04)、(0.29±0.04)](均P<0.05).双荧光素酶靶标实验提示miR-135b-5p靶向FEN1.结论 姜黄素可增强OVCAR-3/DDP对顺铂的敏感性,可能与促进miR-135b-5p的表达从而靶向抑制FEN1的表达相关.
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