长链非编码 RNA CYTOR 通过 miR-136-5p/MAT2B 轴调节肾细胞癌的进展。

Long non-coding RNA CYTOR modulates cancer progression through miR-136-5p/MAT2B axis in renal cell carcinoma.

机构信息

Department of Urology Surgery, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China.

出版信息

Toxicol Appl Pharmacol. 2022 Jul 15;447:116067. doi: 10.1016/j.taap.2022.116067. Epub 2022 May 18.

DOI:10.1016/j.taap.2022.116067

PMID:35597301

Abstract

BACKGROUND

To explore the role of long noncoding RNAs (lncRNAs) cytoskeleton regulator RNA (CYTOR) in renal cell carcinoma (RCC).

METHODS

The levels of CYTOR in RCC tissues and cell lines were detected by RT-qPCR. 786-O and Caki-1 cells were transfected with CYTOR-shRNA or pcDNA-CYTOR respectively, or co-transfected with CYTOR-shRNA and miR-136-5p inhibitor, or co-transfected with miR-136-5p mimic and pcDNA-MAT2B. MTT assay, Transwell assay and flow cytometry were used to evaluate cell proliferation, invasion and apoptosis. The relationship between lncRNA CYTOR and miRNA-136-5p was detected by dual luciferase reporter gene and RNA pull down assays, and the targeted relationship between miRNA-136-5p and MAT2B was verified by dual luciferase reporter gene assay. The interaction between MAT2B and BAG3 protein was verified by co-IP experiment. The role of lncRNA CYTOR in vivo was also examined.

RESULTS

LncRNA CYTOR was up-regulated in RCC tissues and cell lines, and miR-136-5p was down-regulated in renal carcinoma cell lines and tissues. Downregulation of CYTOR inhibited cell proliferation and invasion and promoted apoptosis. miR-136-5p was sponged by lncRNA CYTOR, which negatively regulated the development of RCC. MAT2B was a target gene of miR-136-5p. MAT2B protein interacted directly with BAG3 protein to affect the proliferation, invasion and apoptosis of RCC cells. In vivo experiments showed that the expression level of miR-136-5p was increased, and MAT2B expression was decreased after CYTOR knockdown, thereby inhibiting the development of RCC.

CONCLUSIONS

LncRNA CYTOR promoted the progression of RCC by targeting miR-136-5p to regulate the target gene MAT2B, which interacted with BAG3 protein.

摘要

背景

探讨长链非编码 RNA（lncRNA）细胞骨架调节 RNA（CYTOR）在肾细胞癌（RCC）中的作用。

方法

通过 RT-qPCR 检测 RCC 组织和细胞系中 CYTOR 的水平。分别用 CYTOR-shRNA 或 pcDNA-CYTOR 转染 786-O 和 Caki-1 细胞，或共转染 CYTOR-shRNA 和 miR-136-5p 抑制剂，或共转染 miR-136-5p 模拟物和 pcDNA-MAT2B。MTT 测定、Transwell 测定和流式细胞术用于评估细胞增殖、侵袭和凋亡。通过双荧光素酶报告基因和 RNA 下拉测定检测 lncRNA CYTOR 与 miRNA-136-5p 的关系，并通过双荧光素酶报告基因测定验证 miRNA-136-5p 与 MAT2B 的靶向关系。通过 co-IP 实验验证 MAT2B 和 BAG3 蛋白之间的相互作用。还检查了 lncRNA CYTOR 在体内的作用。

结果

在 RCC 组织和细胞系中上调了 lncRNA CYTOR，而在肾癌细胞系和组织中下调了 miR-136-5p。下调 CYTOR 抑制细胞增殖和侵袭，促进凋亡。miR-136-5p 被 lncRNA CYTOR 海绵吸附，负调控 RCC 的发生。MAT2B 是 miR-136-5p 的靶基因。MAT2B 蛋白与 BAG3 蛋白直接相互作用，影响 RCC 细胞的增殖、侵袭和凋亡。体内实验表明，miR-136-5p 的表达水平升高，而 CYTOR 敲低后 MAT2B 表达降低，从而抑制了 RCC 的发生。