Shi Liangpan, Su Yibin, Zheng Zhihua, Qi Jinyu, Wang Weidong, Wang Cunchuan
Department of Gastrointestinal Surgery, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China.
Department of Gastrointestinal Surgery, The First Hospital of Quanzhou Affiliated of Fujian Medical University, Quanzhou, Fujian 362000, P.R. China.
Exp Ther Med. 2022 Mar;23(3):231. doi: 10.3892/etm.2022.11155. Epub 2022 Jan 20.
Increasing evidence highlights the multiple roles of microRNAs (miRs) in the tumorigenesis of colorectal cancer (CRC); however, the molecular mechanism, particularly the target of miR-146b-5p in CRC has not been fully elucidated. The present study aimed to elucidate the influence of miR-146b-5p via regulating tumor necrosis factor receptor-associated factor 6 (TRAF6) in CRC. The expression levels of miR-146b-5p and TRAF6 in CRC tissue and cells were determined by reverse transcription quantitative PCR and western blotting. Binding between miR-146b-5p and TRAF6 was examined using a dual luciferase reporter gene assay. The impact of miR-146b-5p and TRAF6 on proliferation and migration of CRC cells was determined using Cell Counting Kit-8 and Transwell assays, respectively. An animal model of CRC was established to determine the carcinogenic effect of the miR-146b-5p-TRAF6 axis. The results demonstrated that miR-146b-5p was highly expressed in CRC tissue samples compared with in normal adjacent tissue samples and in CRC cells compared with in the normal NCM460 cell line, whereas TRAF6 was expressed at low levels. Overexpression of miR-146b-5p decreased TRAF6 expression in CRC HT29 and SW620 cells. miR-146b-5p targeted and inhibited TRAF6 expression in CRC cells. Furthermore, transfection with a miR-146b-5p mimic promoted the proliferation, migration and invasion of CRC cells and tumor growth; however, these effects were abolished by TRAF6 overexpression. Transfection with a miR-146b-5p inhibitor suppressed the proliferation of CRC cells. Taken together, the results from the present study demonstrated that miR-146b-5p could enhance the initiation and tumorigenesis of CRC by targeting TRAF6. These results will help elucidate the mechanisms underlying CRC development and will facilitate the development of targeted therapy for CRC.
越来越多的证据凸显了微小RNA（miR）在结直肠癌（CRC）肿瘤发生中的多重作用；然而，其分子机制，尤其是miR-146b-5p在CRC中的靶点尚未完全阐明。本研究旨在通过调控CRC中的肿瘤坏死因子受体相关因子6（TRAF6）来阐明miR-146b-5p的影响。通过逆转录定量PCR和蛋白质印迹法测定CRC组织和细胞中miR-146b-5p和TRAF6的表达水平。使用双荧光素酶报告基因测定法检测miR-146b-5p与TRAF6之间的结合。分别使用细胞计数试剂盒-8和Transwell测定法确定miR-146b-5p和TRAF6对CRC细胞增殖和迁移的影响。建立CRC动物模型以确定miR-146b-5p-TRAF6轴的致癌作用。结果表明，与正常相邻组织样本相比，miR-146b-5p在CRC组织样本中高表达，与正常NCM460细胞系相比，在CRC细胞中高表达，而TRAF6表达水平较低。miR-146b-5p的过表达降低了CRC HT29和SW620细胞中TRAF6的表达。miR-146b-5p靶向并抑制CRC细胞中TRAF6的表达。此外，用miR-146b-5p模拟物转染促进了CRC细胞的增殖、迁移和侵袭以及肿瘤生长；然而，TRAF6过表达消除了这些作用。用miR-146b-5p抑制剂转染抑制了CRC细胞的增殖。综上所述，本研究结果表明，miR-146b-5p可通过靶向TRAF6增强CRC的起始和肿瘤发生。这些结果将有助于阐明CRC发展的潜在机制，并将促进CRC靶向治疗的发展。
