Yu Jie, Tan Qunyou, Deng Bo, Fang Chunshu, Qi Di, Wang Ruwen
Department of Thoracic Surgery, Institute of Surgery Research, Daping Hospital, The Third Military Medical University Chongqing 400038, P. R. China.
Am J Cancer Res. 2015 Jan 15;5(2):802-11. eCollection 2015.
Growing evidence indicates that miR-520a was involved in the complement attack and migration of tumor cells, but nonetheless, the role of miR-520a-3p in non-small cell lung cancer (NSCLC) is not clear. Mitogen-activated protein kinase kinase kinase 2 (MAP3K2) is a kinase belonging to the serine/threonine protein kinase family. To develop potential therapy targeting MAP3K2, we studied the roles of miR-520a-3p in the proliferation, apoptosis and metastasis of NSCLC. The expression levels of miR-520a-3p were quantified in tumor tissues of NSCLC by qRT-PCR, and the mimics and inhibitors were used to verify the function of miR-520a-3p. The cell proliferation was evaluated by MTT assay, and the migration and invasion was evaluated by transwell assay. The athymic mice subcutaneous injection was used to research NSCLC cell tumor formation. The bioinformatics tools and luciferase assay was applied to detect the relationship between miR-520a-3p and its target. Protein levels of miR-520a-3p target was determined by western blot analysis. MiR-520a-3p expression was decreased in the NSCLC tissues compared with their normal counterparts and lower expression of miR-520a-3p in NSCLC tissues was associated with a higher clinical stage, NSCLC metastasis and poor prognosis. Inhibition of expression of miR-520a-3p can reduce in vitro NSCLC cell migration and invasion as well as in vivo metastasis. MAP3K2 mRNA contains a binding site for miR-520a-3p in the 3'UTR. MAP3K2 is one of target of miR-520a-3p. Together, our data demonstrated that miR-520a-3p inhibits proliferation, apoptosis and metastasis in NSCLC by targeting MAP3K2, and miR-520a-3p may be used as a prognosis marker for NSCLC in clinical research.
越来越多的证据表明,miR-520a参与了肿瘤细胞的补体攻击和迁移,但miR-520a-3p在非小细胞肺癌(NSCLC)中的作用尚不清楚。丝裂原活化蛋白激酶激酶激酶2(MAP3K2)是一种属于丝氨酸/苏氨酸蛋白激酶家族的激酶。为了开发针对MAP3K2的潜在治疗方法,我们研究了miR-520a-3p在NSCLC增殖、凋亡和转移中的作用。通过qRT-PCR对NSCLC肿瘤组织中miR-520a-3p的表达水平进行定量,并使用模拟物和抑制剂来验证miR-520a-3p的功能。通过MTT法评估细胞增殖,通过Transwell法评估迁移和侵袭。采用无胸腺小鼠皮下注射法研究NSCLC细胞肿瘤形成。应用生物信息学工具和荧光素酶测定法检测miR-520a-3p与其靶标的关系。通过蛋白质印迹分析确定miR-520a-3p靶标的蛋白质水平。与正常组织相比,NSCLC组织中miR-520a-3p表达降低,NSCLC组织中miR-520a-3p低表达与更高的临床分期、NSCLC转移和不良预后相关。抑制miR-520a-3p的表达可降低体外NSCLC细胞的迁移和侵袭以及体内转移。MAP3K2 mRNA在3'UTR中含有miR-520a-3p的结合位点。MAP3K2是miR-520a-3p的靶标之一。总之,我们的数据表明,miR-520a-3p通过靶向MAP3K2抑制NSCLC的增殖、凋亡和转移,并且miR-520a-3p可能在临床研究中用作NSCLC的预后标志物。