Liu Yong-Gang, Li Jia, Nie Fang, Jin Gao-Wa
Department of Thoracic Oncology, Baotou Cancer Hospital Baotou 014000, Inner Mongolia, China.
Department of Immunization Planning, Baotou Disease Control and Defense Center Baotou 014000, Inner Mongolia, China.
Am J Transl Res. 2022 Feb 15;14(2):888-898. eCollection 2022.
This study set out to explore the regulatory relationship between LINC00961/miR-3127 axis and non-small-cell lung carcinoma (NSCLC), so as to provide a new and effective molecular target for targeted therapy of NSCLC.
RNA-seq and miRNA-seq data of NSCLC and normal samples were obtained from The Cancer Genome Atlas (TCGA) database for analyzing LINC00961 and miR-3127 expression. Eighty-six pairs of clinical NSCLC tissues and adjacent normal tissues as well as NSCLC cell lines were obtained. Measurements of LINK00961 and miR-3127 levels were done using real-time-quantitative polymerase chain reaction (RT-qPCR). Furthermore, LINK00961 and miR-3127 in NSCLC cell were regulated respectively. The NSCLC cell proliferation, invasion and migration were determined with MTT assay, Transwell and wound healing assays, respectively. The levels of invasion- and apoptosis-related proteins were detected using western blots, and the connection of LINC00961 and miR-3127 was identified using dual luciferase reporter (DLR) assay.
Differential analysis results of TCGA databases identified that LINC00961 was ubiquitously expressed at low levels in NSCLC, while miR-3127 was highly expressed. Similar expression trends of LINC00961 and miR-3127 were observed in clinical NSCLC samples and cell lines. Overexpression of LINC00961 and knockdown of miR-3127 significantly reduced NCI-H1299 cell migration, invasiveness, and multiplication, decreased MMP-2, MMP-9 and Bcl-2 protein levels, and increased E-cadherin, Bax and Caspase-3 protein levels. The DLR assay confirmed that miR-3127 can be targeted by LINC00961.
LINC00961 functions as an anti-oncogene in NSCLC by modulating miR-3127.
本研究旨在探讨LINC00961/miR - 3127轴与非小细胞肺癌（NSCLC）之间的调控关系，从而为NSCLC的靶向治疗提供新的有效分子靶点。
从癌症基因组图谱（TCGA）数据库获取NSCLC和正常样本的RNA测序和miRNA测序数据，以分析LINC00961和miR - 3127的表达。获取86对临床NSCLC组织及相邻正常组织以及NSCLC细胞系。使用实时定量聚合酶链反应（RT - qPCR）测量LINK00961和miR - 3127水平。此外，分别对NSCLC细胞中的LINK00961和miR - 3127进行调控。分别采用MTT法、Transwell法和伤口愈合试验测定NSCLC细胞的增殖、侵袭和迁移能力。使用蛋白质免疫印迹法检测侵袭和凋亡相关蛋白的水平，并使用双荧光素酶报告基因（DLR）试验鉴定LINC00961和miR - 3127的关联。
TCGA数据库的差异分析结果表明，LINC00961在NSCLC中普遍低表达，而miR - 3127高表达。在临床NSCLC样本和细胞系中观察到LINC00961和miR - 3127相似的表达趋势。LINC00961的过表达和miR - 3127的敲低显著降低了NCI - H1299细胞的迁移、侵袭和增殖能力，降低了MMP - 2、MMP - 9和Bcl - 2蛋白水平，并增加了E - cadherin、Bax和Caspase - 3蛋白水平。DLR试验证实miR - 3127可被LINC00961靶向。
LINC00961通过调节miR - 3127在NSCLC中发挥抑癌基因的作用。
