Dong Yu, Huo Xiaoying, Sun Ruiying, Liu Zhiyan, Huang Miaoyi, Yang Shuanying
Department of Respiratory Medicine, Xi'an Central Hospital, Xi'an, China.
Department of Respiratory Medicine, Fourth Hospital of Xi'an, Xi'an, China.
Oncol Res. 2017 May 5. doi: 10.3727/096504017X14930316817366.
Long non-coding RNAs (lncRNAs) have been involved in occurrence and progression of multiple cancers. In this study, weinvestigated the role of lncRNA Gm15290 in the proliferation and invasion of non-small cell lung cancer (NSCLC) cells. First, we found that lncRNA Gm15290 was markedly upregulated in tumor tissues from NSCLC patients and NSCLC cell lines, compared with adjacent normal tissues and normal lung cell line HBE respectively. Then, different concentrations of pcDNA-Gm15290 expression vector and Gm15290 siRNA were respectively transfected into A549 NSCLC cells. Our results showed that overexpression of Gm15290 significantly increased the proliferation and invasion of A549 cells, and suppressed cell apoptosis. Knockdown of Gm15290 suppressed A549 cell proliferation and invasion, and promoted cell apoptosis. Subsequently, we explored the underlying mechanism through which Gm15290 promoted cell proliferation and invasion. The output of RNAhybrid bioinformatic tool revealed that Gm15290 potentially interacted with tumor suppressor miR-615-5p which displayed an opposite expression pattern in the cell lines and a strong negative correlation with the levels of Gm15290 in NSCLC patients (r = 0.9677, P < 0.0001). The results of RNA pull-down assays confirmed that Gm15290 directly bound with miR-615-5p. Gm15290 negatively regulated the expression of miR-615-5p and increased the protein levels of miR-615-5p target genes, including IGF2, AKT2 and SHMT2. Moreover, miR-615-5p mimic could antagonize the promoting effect of Gm15290 on cell proliferation and invasion.
长链非编码RNA（lncRNAs）参与了多种癌症的发生和发展。在本研究中，我们调查了lncRNA Gm15290在非小细胞肺癌（NSCLC）细胞增殖和侵袭中的作用。首先，我们发现与相邻正常组织和正常肺细胞系HBE相比，lncRNA Gm15290在NSCLC患者的肿瘤组织和NSCLC细胞系中显著上调。然后，将不同浓度的pcDNA-Gm15290表达载体和Gm15290 siRNA分别转染到A549 NSCLC细胞中。我们的结果表明，Gm15290的过表达显著增加了A549细胞的增殖和侵袭，并抑制了细胞凋亡。敲低Gm15290可抑制A549细胞的增殖和侵袭，并促进细胞凋亡。随后，我们探索了Gm15290促进细胞增殖和侵袭的潜在机制。RNAhybrid生物信息学工具的输出结果显示，Gm15290可能与肿瘤抑制因子miR-615-5p相互作用，miR-615-5p在细胞系中呈现相反的表达模式，且与NSCLC患者中Gm15290的水平呈强负相关（r = 0.9677，P < 0.0001）。RNA下拉实验结果证实Gm15290直接与miR-615-5p结合。Gm15290负向调节miR-615-5p的表达，并增加了miR-615-5p靶基因的蛋白水平，包括IGF2、AKT2和SHMT2。此外，miR-615-5p模拟物可拮抗Gm15290对细胞增殖和侵袭的促进作用。
