Su Hongbo, Fan Guanzhi, Huang Jin, Qiu Xueshan
Department of Pathology, First Affiliated Hospital, China Medical University, Shenyang 110001, Liaoning Province, PR China.
Department of Pathology, Shengjing Affiliated Hospital, China Medical University, Shenyang 110004, Liaoning Province, PR China.
Cell Signal. 2021 Sep;85:110049. doi: 10.1016/j.cellsig.2021.110049. Epub 2021 May 31.
Y-box binding protein 1 (YBX1) is a common oncogene in non-small-cell lung cancer (NSCLC), which is regulated by microRNAs (miRNAs) and transcription factors. This research aims to explore the function of YBX1, miR-148a-3p and Runt-related transcription factor 3 (Runx3) in NSCLC development, and analyze their interactions.
YBX1, miR-148a-3p and Runx3 levels were detected using quantitative reverse transcription polymerase chain reaction(RT-PCR), Western blotting or immunohistochemical staining. The functions of YBX1, miR-148a-3p and Runx3 were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, wound healing, transwell, flow cytometry, xenograft model and Western blotting analyses. The binding correlation was validated through dual-luciferase reporter analysis and chromatin immunoprecipitation (ChIP).
YBX1 expression was upregulated, and miR-148a-3p and Runx3 levels were reduced in NSCLC samples and cell lines. YBX1 silence restrained NSCLC cell proliferation, migration, invasion and tumor growth, and enhanced apoptosis. YBX1 was targeted via miR-148a-3p. MiR-148a-3p knockdown promoted cell proliferation, migration, invasion and tumor growth, and repressed apoptosis, and these effects were abolished by YBX1 silence. Runx3 upregulation restrained cell proliferation, migration, invasion and tumor growth, and facilitated apoptosis. Runx3 bound with miR-148a-3p promotor to regulate miR-148a-3p expression. Runx3 silence modulated YBX1 expression though miR-148a-3p to promote NSCLC progression by increasing Cyclin D1, Cyclin B1, Slug-1, MMP-2 and MMP-9 levels.
Runx3-miR-148a-3p axis targeted YBX1 to modulate NSCLC progression.
Y盒结合蛋白1（YBX1）是非小细胞肺癌（NSCLC）中一种常见的癌基因，其受微小RNA（miRNA）和转录因子调控。本研究旨在探讨YBX1、miR-148a-3p和 runt相关转录因子3（Runx3）在NSCLC发生发展中的作用，并分析它们之间的相互作用。
采用定量逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法或免疫组织化学染色检测YBX1、miR-148a-3p和Runx3的水平。通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）、集落形成、伤口愈合、Transwell、流式细胞术、异种移植模型和蛋白质免疫印迹分析评估YBX1、miR-148a-3p和Runx3的功能。通过双荧光素酶报告基因分析和染色质免疫沉淀（ChIP）验证结合相关性。
NSCLC样本和细胞系中YBX1表达上调，miR-148a-3p和Runx3水平降低。YBX1沉默抑制NSCLC细胞增殖、迁移、侵袭和肿瘤生长，并增强细胞凋亡。YBX1是miR-148a-3p的靶标。miR-148a-3p敲低促进细胞增殖、迁移、侵袭和肿瘤生长，并抑制细胞凋亡，而YBX1沉默可消除这些作用。Runx3上调抑制细胞增殖、迁移、侵袭和肿瘤生长，并促进细胞凋亡。Runx3与miR-148a-3p启动子结合以调节miR-148a-3p表达。Runx3沉默通过miR-148a-3p调节YBX1表达，通过增加细胞周期蛋白D1、细胞周期蛋白B1、Slug-1、基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）水平促进NSCLC进展。
Runx3-miR-148a-3p轴靶向YBX1调节NSCLC进展。
