Guo Shuai, Li Ming, Li Juan, Lv Yan
Department of Medical Oncology, Shandong Provincial Chest Hospital, Jinan, Shandong 250013, P.R. China.
Department of Thoracic Surgery, Shandong Provincial Chest Hospital, Jinan, Shandong 250013, P.R. China.
Oncol Lett. 2020 Feb;19(2):1516-1522. doi: 10.3892/ol.2019.11194. Epub 2019 Dec 10.
Effect of targeted regulation of mothers against decapentaplegic homolog 3 (Smad3) by microRNA-15a (miR-15a) on the proliferation, invasion and metastasis of non-small cell lung cancer (NSCLC) cells and its related mechanisms were investigated. Fifty pairs of NSCLC and para-cancerous tissues were collected to identify the expression level of miR-15a in NSCLC, para-cancerous tissue, and cell lines A549, H1299, H1975 and BEAS-2B by real-time fluorescence quantitative PCR (RT-PCR); A549 cells were transfected with miR-15a mimic; the MTT assay was performed to detect the role of miR-15a transfection in proliferation of A549 cells, the wound healing assay was carried out to identify the role of miR-15a in migration of A549 cells; Transwell invasion assay was conducted to analyze the role of miR-15a in invasion of A549 cells; western blotting was carried out to find the effect of miR-15a on Smad3 expression, and Spearman's rank correlation was used to analyze the correlation between miR-15a and Smad3 expression. NSCLC tissues and cells showed significantly lower miR-15a expression, compared with para-cancerous tissues and normal cell lines (P=0.023). miR-15a was significantly more expressed in A549 cells transfected with miR-15a mimic (P=0.043). Overexpression of miR-15a can significantly inhibit A549 cell proliferation (P=0.038), migration (P=0.033) and invasion (P=0.025), and significantly reduced the expression level of Smad3 (P=0.031) in A549 cells. Spearman's rank correlation showed negative correlation of miR-15a expression with Smad3, which may indicate negative regulation (r=-0.34, P<0.0001). Inhibition of proliferation, migration and invasion of NSCLC cells can be achieved with targeted regulation of Smad3 by miR-15a.
研究了微小RNA-15a（miR-15a）对母亲针对截瘫同源物3（Smad3）的靶向调控对非小细胞肺癌（NSCLC）细胞增殖、侵袭和转移的影响及其相关机制。收集50对NSCLC组织和癌旁组织，通过实时荧光定量PCR（RT-PCR）检测miR-15a在NSCLC组织、癌旁组织以及细胞系A549、H1299、H1975和BEAS-2B中的表达水平；用miR-15a模拟物转染A549细胞；采用MTT法检测miR-15a转染对A549细胞增殖的作用，进行伤口愈合试验以确定miR-15a在A549细胞迁移中的作用；进行Transwell侵袭试验分析miR-15a在A549细胞侵袭中的作用；进行蛋白质印迹法以发现miR-15a对Smad3表达的影响，并采用Spearman等级相关分析miR-15a与Smad3表达之间的相关性。与癌旁组织和正常细胞系相比，NSCLC组织和细胞中miR-15a表达显著降低（P=0.023）。在转染miR-15a模拟物的A549细胞中，miR-15a表达显著增加（P=0.043）。miR-15a过表达可显著抑制A549细胞增殖（P=0.038）、迁移（P=0.033）和侵袭（P=0.025），并显著降低A549细胞中Smad3的表达水平（P=0.031）。Spearman等级相关显示miR-15a表达与Smad3呈负相关，这可能表明存在负调控（r=-0.34，P<0.0001）。通过miR-15a对Smad3进行靶向调控可实现对NSCLC细胞增殖、迁移和侵袭的抑制。
