摘要： 目的 探讨长链非编码RNA （lncRNA） MALAT1靶向miR-218对卵巢癌细胞系OVCAR3增殖、侵袭及凋亡的影响及作用机制。方法 应用实时PCR检测lncRNA MALAT1和miR-218在人卵巢癌细胞系 （OVCAR3） 和正常人卵巢上皮细胞系 （HOSE） 中的表达。利用脂质体转染法将无意义序列阴性对照 （si-NC） 组、MALAT1干扰序列 （si-MALAT1） 组以及si-MALAT1+miR-218抑制剂组分别转染卵巢癌细胞，MTT法检测各组细胞增殖情况，流式细胞仪检测各组细胞凋亡情况，Transwell 实验检测各组细胞侵袭情况。生物信息学分析及双荧光素酶报告基因实验验证MALAT1和miR-218的靶向关系。实时PCR检测各组OVCAR3细胞中MALAT1、miR-218的表达； Western blotting检测各组OVCAR3细胞中Runx2的表达。结果 与人正常卵巢上皮细胞比较，卵巢癌OVCAR3细胞中MALAT1表达升高 （P< 0.05），miR-218表达下降 （P< 0.05）。与si-NC组比较，si-MALAT1组OVCAR3细胞的增殖和侵袭能力下降，凋亡率升高 （均P< 0.05）。与si-MALAT组比较，si-MALAT1+miR-218抑制剂组OVCAR3细胞的增殖和侵袭能力升高，凋亡率下降 （均P< 0.05）。与si-NC组比较，si-MALAT1组OVCAR3细胞中miR-218表达升高 （P< 0.05）。生物信息学分析和双荧光素酶报告基因实验结果显示MALAT1可以靶向调控miR-218。Western blotting结果显示，与 si-NC组比较，si-MALAT1组Runx2蛋白表达降低 （P< 0.05），与si-MALAT1组比较，si-MALAT1+miR-218抑制剂组Runx2蛋白表达水平升高 （P< 0.05）。结论 lncRNA MALAT1在卵巢癌细胞中高表达，抑制MALAT1表达能够抑制卵巢癌OVCAR3细胞的增殖和侵袭，并促进OVCAR3细胞凋亡，其机制可能与通过miR-218调控Runx2蛋白表达有关。

长链非编码RNA MALAT1靶向miR-218对卵巢癌细胞增殖、侵袭及凋亡的影响及其作用机制

邱辉, 周佳任

中国医科大学附属盛京医院妇产科, 沈阳 110004

收稿日期:2022-11-19出版日期:2023-05-30发布日期:2023-05-26
通讯作者:周佳任E-mail:zhoujr2013sj@163.com
作者简介:邱辉 (1983-),女,讲师,博士.



关键词: 长链非编码RNA, MALAT1, 微RNA 218, 卵巢癌, 增殖, 侵袭, 凋亡
Abstract: Objective To investigate the effect of long noncoding RNA (lncRNA) MALAT1 on proliferation,invasion,and apoptosis of ovarian cancer cell OVCAR3 by targeting microRNA 218 (miR-218) and its mechanism. Methods MALAT1 and miR-218 expressions in ovarian cancer cells and normal human ovarian epithelial cells were detected by qRT-PCR. Small interfering RNA negative control (si-NC) group,silencing MALAT1 (si-MALAT1) group,and si-MALAT1 and miR-218 inhibitor group were transfected in OVCAR3 by lipofection. Cell viability,cell invasion,and cell apoptosis were detected by MTT,the Transwell experiment,and flow cytometry,respectively. Biological website analysis and double luciferase reporter gene experiment were used to study the targeting relationship between MALAT1 and miR-218. MALAT1 and miR-218 expressions were detected by qRT-PCR,and Runx2 expression was detected by Western blotting. Results Compared with human normal ovarian epithelial cell HOSE,MALAT1 expression increased,whereas miR-218 expression decreased in ovarian cancer OVCAR3 cells (P< 0.05). After transfection of si-MALAT1,the proliferation and invasion ability of OVCAR3 ovarian cancer cells decreased,and the apoptosis rate increased (P< 0.05). Bioinformatics and luciferase analysis showed that MALAT1 could specifically regulate miR-218. Western blotting results showed that Runx2 expression in the si-MALAT1 group was lower than that in the si-NC group,whereas Runx2 protein expression in the si-MALAT1 + miR-218 inhibitor group was higher than that in the si-MALAT1 group (P< 0.05). Conclusion lncRNA MALAT1 is highly expressed in ovarian cancer cells. The silence of lncRNA MALAT1 can inhibit the proliferation and invasion of ovarian cancer cells OVCAR3 and promote its apoptosis. The mechanism may be related to the regulation of Runx2 protein expression by miR-218.
Key words: long noncoding RNA, MALAT1, microRNA 218, ovarian cancer, proliferation, invasion, apoptosis
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