敲减lncRNA RP11-626G11.3通过调控miR-532-3p抑制人肾癌细胞系的增殖和迁移

doi:10.16352/j.issn.1001-6325.2023.10.1498

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (10): 1498-1504.doi: 10.16352/j.issn.1001-6325.2023.10.1498

敲减lncRNA RP11-626G11.3通过调控miR-532-3p抑制人肾癌细胞系的增殖和迁移

王文龙¹, 唐一君^2*, 王青兵¹, 陈科³, 程继强¹

河南科技大学附属安阳市肿瘤医院 1.外科五病区;
2.肿瘤内科五病区,河南安阳 455000;
3.华中科技大学附属协和医院肾内科,湖北武汉 430022

收稿日期:2023-01-17 修回日期:2023-07-18 出版日期:2023-10-05 发布日期:2023-10-05
通讯作者: *tyjnanyang@163.com
基金资助:
国家自然科学基金(81772721)

Knockdown of lncRNA RP11-626G11.3 inhibits proliferation and migration of human renal carcinoma cell lines through regulating miR-532-3p

WANG Wenlong¹, TANG Yijun^2*, WANG Qingbing¹, CHEN Ke³, CHENG Jiqiang¹

1. Department of the Fifth Ward of Surgery;
2. Department of the Fifth Ward of Medical Oncology, Anyang Cancer Hospital Affiliated to Henan University of Science and Technology, Anyang 455000;
3. Department of Nephrology, Union Hospital Affiliated to Huazhong University of Science and Technology, Wuhan 430022, China

Received:2023-01-17 Revised:2023-07-18 Online:2023-10-05 Published:2023-10-05
Contact: *tyjnanyang@163.com

摘要/Abstract

摘要： 目的研究长链非编码RNA(lncRNA) RP11-626G11.3在肾癌组织和细胞系中的表达,探究敲减RP11-626G11.3对肾癌细胞生物学行为的影响。方法用GEPIA数据库分析RP11-626G11.3在肾癌组织和癌旁组织中的表达情况,用TCGA数据库分析RP11-626G11.3表达水平与肾癌患者生存期的关系。QPCR检测RP11-626G11.3在多种肾癌细胞系中的表达。选择RP11-626G11.3表达最高的肾癌细胞系,分别转染对照质粒(si-NC组)和RP11-626G11.3沉默质粒(si-RP11-626G11.3组)。MTT法和细胞划痕实验检测细胞的增殖和迁移。通过生物信息学方法找到与RP11-626G11.3结合的微小RNA(miRNA),并用双荧光素酶报告实验进行验证。QPCR检测两组细胞中miR-532-3p的表达。Western blot检测两组细胞中Wnt/β-catenin信号通路蛋白的表达。结果与癌旁组织相比,肾癌组织中RP11-626G11.3 表达量上调(P＜0.01)。与RP11-626G11.3高表达的患者相比,RP11-626G11.3低表达的患者生存期较长(P＜0.01)。与永生化肾小管上皮细胞相比,肾癌细胞系中RP11-626G11.3表达量均上调(P＜0.01),OS-RC-2细胞中RP11-626G11.3的表达量最高(P＜0.01)。与si-NC组相比,si-RP11-626G11.3组OS-RC-2细胞活力明显降低(P＜0.05),细胞迁移率明显降低(P＜0.01)。RP11-626G11.3靶向结合miR-532-3p(P＜0.01)。相比si-NC组,si-RP11-626G11.3组OS-RC-2细胞中miR-532-3p表达明显上调(P＜0.01),Wnt/β-catenin信号通路蛋白β-catenin、Axin2、C-myc、cyclin D1、MMP7表达水平下降。结论 RP11-626G11.3在肾癌组织和细胞系中表达量升高,敲减RP11-626G11.3通过调控miR-532-3p表达抑制肾癌细胞系的增殖和迁移。

关键词: 肾肿瘤, RP11-626G11.3, miR-532-3p, 增殖, 迁移

Abstract: Objective To study the expression of long-chain non-coding RNA (lncRNA) RP11-626G11.3 in renal cancer tissues and cell lines, and to explore the effect of knockdown of RP11-626G11.3 on the biological behavior of renal cancer cells. Methods The GEPIA database was used to analyze the expression of RP11-626G11.3 in renal cancer tissues and adjacent tissues, and the TCGA database was used to analyze the relationship between the expression of RP11-626G11.3 and the survival time of renal cancer patients. The expression of RP11-626G11.3 in various renal cancer cell lines was detected by qPCR. The renal cancer cells with the highest expression of RP11-626G11.3 were selected and transfected with control plasmid (si-NC group) and RP11-626G11.3 silencing plasmid (si-RP11-626G11.3 group). Cell proliferation and migration were examined by MTT assay and cell scratch assay. The microRNA (miRNA) binding to RP11-626G11.3 was found by bioinformatics method, and verified by dual-luciferase reporter gene experiment. The expression of miR-532-3p in the two groups of cells was detected by qPCR. Western blot was used to detect the expression levels of Wnt/β-catenin signaling pathway in the two groups of cells. Results Compared with adjacent tissues, the expression of RP11-626G11.3 was up-regulated in renal cancer tissues (P＜0.01). Compared with patients with high expression of RP11-626G11.3, patients with low expression of RP11-626G11.3 had a longer survival time (P＜0.01). Compared with immortalized renal tubular epithelial cells, the expression of RP11-626G11.3 was up-regulated in renal cancer cell lines (P＜0.01), and the expression of RP11-626G11.3 was the highest in OS-RC-2 cells (P＜0.01). Compared with si-NC group, the viability of OS-RC-2 cells in si-RP11-626G11.3 group was significantly decreased (P＜0.05) with decreased cell migration rate(P＜0.01). RP11-626G11.3 was found to target at miR-532-3p (P＜0.01). Compared with the si-NC group, the expression of miR-532-3p in OS-RC-2 cells in the si-RP11-626G11.3 group was significantly up-regulated (P＜0.01), and the Wnt/β-catenin signaling pathway proteins β-catenin, Axin2, C-myc, cyclin D1 and MMP7 decreased. Conclusions The expression of RP11-626G11.3 is increased in renal cancer tissues and cell lines. Knockdown of RP11-626G11.3 inhibits the proliferation and migration of renal cancer cells by regulating the expression of miR-532-3p.

Key words: renal tumor, RP11-626G11.3, miR-532-3p, proliferation, migration

中图分类号:

R737.11

王文龙, 唐一君, 王青兵, 陈科, 程继强. 敲减lncRNA RP11-626G11.3通过调控miR-532-3p抑制人肾癌细胞系的增殖和迁移[J]. 基础医学与临床, 2023, 43(10): 1498-1504.

WANG Wenlong, TANG Yijun, WANG Qingbing, CHEN Ke, CHENG Jiqiang. Knockdown of lncRNA RP11-626G11.3 inhibits proliferation and migration of human renal carcinoma cell lines through regulating miR-532-3p[J]. Basic & Clinical Medicine, 2023, 43(10): 1498-1504.

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敲减lncRNA RP11-626G11.3通过调控miR-532-3p抑制人肾癌细胞系的增殖和迁移

Knockdown of lncRNA RP11-626G11.3 inhibits proliferation and migration of human renal carcinoma cell lines through regulating miR-532-3p

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