miR-146a-5p靶向SMAD4抑制人前列腺癌细胞系PC-3增殖和侵袭

doi:10.16352/j.issn.1001-6325.2023.08.1215

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (8): 1215-1221.doi: 10.16352/j.issn.1001-6325.2023.08.1215

miR-146a-5p靶向SMAD4抑制人前列腺癌细胞系PC-3增殖和侵袭

刘彼得, 李循, 王书恒, 靳宏勇, 张小安, 李九智^*

新疆维吾尔自治区人民医院泌尿中心泌尿外科研究室,新疆乌鲁木齐 830001

收稿日期:2022-08-20 修回日期:2023-01-06 出版日期:2023-08-05 发布日期:2023-07-26
通讯作者: ^*xjlijiuzhi@163.com
基金资助:
新疆维吾尔自治区科学技术厅自治区自然科学基金(2017D01C102)

miR-146a-5p inhibits proliferation and invasion of prostate cancer cell line PC-3 by targeting SMAD4

LIU Bide, LI Xun, WANG Shuheng, JIN Hongyong, ZHANG Xiao'an, LI Jiuzhi^*

Department of Urology, Laboratory of Urology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China

Received:2022-08-20 Revised:2023-01-06 Online:2023-08-05 Published:2023-07-26
Contact: ^*xjlijiuzhi@163.com

摘要/Abstract

摘要： 目的探究miR-146a-5p可否靶向调控SMAD4对前列腺癌细胞增殖、侵袭的影响及其机制。方法 RT-qPCR检测前列腺癌组织及各细胞系中miR-146a-5p的表达量,分析其表达与Gleason评分的关系;MTT实验、BrdU实验、集落生成实验、划痕实验、Transwell小室法及裸鼠成瘤实验分析miR-146a-5p对前列腺癌细胞增殖、成瘤、迁移及侵袭等能力的影响;RT-qPCR检测组织SMAD4的表达量,分析其与miR-146a-5p表达量的关系;荧光素酶报告基因实验分析miR-146a-5p与SMAD4的靶向关系,功能回复实验验证miR-146a-5p/SMAD4信号轴在前列腺癌中的作用;Western blot检测miR-146a-5p及SMAD4对细胞核内SMAD2/SMAD3复合体表达量的影响,并通过荧光素酶报告基因实验及染色质免疫共沉淀实验探究miR-146a-5p/SMAD4/SMAD2/SMAD3信号轴对TIM3的靶向调控作用。结果 miR-146a-5p在前列腺癌组织及细胞系中低表达(P＜0.05),其表达量与Gleason评分负相关(P＜0.05),且在PC-3细胞中的表达量最低;miR-146a-5p抑制PC-3细胞的增殖及侵袭能力(P＜0.05);SMAD4为miR-146a-5p的下游靶基因;SMAD4促进SMAD2/SMAD3复合体入核并靶向激活TIM3。结论 miR-146a-5p靶向调控SMAD4抑制PC-3细胞的增殖及侵袭,SMAD2/SMAD3/TIM3信号通路可能为其下游机制。

关键词: 前列腺癌, miR-146a-5p, SMAD4, TIM3

Abstract: Objective To explore the inhibition effect and mechanism of miR-146a-5p on proliferation and invasion of prostate cancer (PCa) cell line PC-3 by targeting SMAD4. Methods RT-qPCR was used to detect the expression of miR-146a-5p in PCa tissues and cell lines. The relevance of miR-146a-5p expression with Gleason score was also analyzed. MTT, BrdU experiment, cell colony formation experiment, scratch experiment, Transwell assay and nude mouse xenograft model experiment were conducted to detect the effect of miR-146a-5p on cell proliferation, tumorigenicity, migration and invasion. The expression of SMAD4 in PCa tissues was detected by RT-qPCR, and the targeting relationship of SMAD4 and miR-146a-5p was confirmed by double luciferase reporter gene assay and rescue experiment. Western blot was used to detect the expression of SMAD2/SMAD3 complex in nucleus affected by miR-146a-5p and SMAD4. Finally, double luciferase reporter gene assay and ChIP experiment were performed to examine the targeting regulation of TIM3 by miR-146a-5p/SMAD4/SMAD2/SMAD3 signaling axis. Results miR-146a-5p was low expressed in PCa tissues and cell lines; its expression was negatively correlated to Gleason score and had the lowest expression in PC-3 cells. miR-146a-5p inhibited the proliferation and invasion of PC-3 cells by targeting SMAD4. SMAD2/SMAD3/TIM3 axis seemed to be the downstream mechanism of miR-146a-5p/SMAD4 signaling pathway. Conclusions miR-146a-5p can inhibit the proliferation and invasion of PC-3 cells by targeting SMAD4, and the downstream mechanism might be related to the SMAD2/SMAD3/TIM3 signaling pathway.

Key words: prostate cancer, miR-146a-5p, SMAD4, TIM3

中图分类号:

R737.25

刘彼得, 李循, 王书恒, 靳宏勇, 张小安, 李九智. miR-146a-5p靶向SMAD4抑制人前列腺癌细胞系PC-3增殖和侵袭[J]. 基础医学与临床, 2023, 43(8): 1215-1221.

LIU Bide, LI Xun, WANG Shuheng, JIN Hongyong, ZHANG Xiao'an, LI Jiuzhi. miR-146a-5p inhibits proliferation and invasion of prostate cancer cell line PC-3 by targeting SMAD4[J]. Basic & Clinical Medicine, 2023, 43(8): 1215-1221.

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miR-146a-5p靶向SMAD4抑制人前列腺癌细胞系PC-3增殖和侵袭

miR-146a-5p inhibits proliferation and invasion of prostate cancer cell line PC-3 by targeting SMAD4

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