Nanjing Medical University
摘 要 目的：探讨长链非编码RNA 01518（long intergenic non-protein coding RNA 01518, LINC01518）基因过表达或沉默对IL-17诱导非小细胞肺癌（non-small cell lung cancer, NSCLC）细胞增殖的影响。方法：在用Western blot检查NSCLC细胞系（PC9、H1299和H1975）IL-17受体A（IL-17 receptor A, IL-17RA）的表达后，行IL-17刺激H1299和PC9细胞不同时间，利用CCK8实验测定细胞的增殖水平。根据LncRNA芯片筛查IL-17刺激两细胞3h时上调LncRNA的结果，从中选取某些上调的LncRNA进行RT-PCR和Real-time PCR验证。此外，将构建的pcDNA3.1/LINC01518或shLINC01518质粒转染H1299细胞，用CCK8、EdU和克隆形成实验检查细胞的增殖情况。结果：3种NSCLC细胞系均有IL-17RA的表达。实验发现，IL-17刺激H1299和PC9细胞后能提高其增殖水平，同时受刺激的细胞内LINC01518上调的最为显著，且过表达LINC01518可提高H1299细胞的增殖水平，而沉默LINC01518基因后则能抑制IL-17诱导的细胞增殖。结论：LINC01518能促进IL-17刺激H1299细胞诱导的细胞增殖。
Abstract Objective: To explore the effect of overexpressing or silencing long intergenic non-protein coding RNA 01518 (LINC01518) on the cell proliferation induced by IL-17 in non-small cell lung cancer (NSCLC). Methods: The IL-17 receptor A (IL-17RA) expression in NSCLC cell lines (PC9, H1299 and H1975) was first examined using Western blot. Then H1299 and PC9 cells were exposed to IL-17 for different time, and the cell proliferation was detected by CCK8 assay. Based on the results of up-regulated lncRNAs detected by lncRNA microarray in IL-17-stimulated H1299 and PC9 cells, we selected and verified several lncRNAs expression level by RT-PCR and Real-time PCR. Additionally, we transfected the constructed plasmids of pcDNA3.1/LINC01518 or shLINC01518 into H1299 cells, and examined the parameters of H1299 cell proliferation by CCK8, EdU and colony formation assays. Results: IL-17RA could express in 3 NSCLC cell lines. The cell proliferation and LINC01518 were obviously up-regulated in H1299 and PC9 cells treated by IL-17. Besides, LINC01518 overexpression could markedly promote the cell proliferation, while LINC01518 gene knockdown followed by IL-17 treatment did not enhance the level of cell proliferation. Conclusion: LINC01518 can promote H1299 cell proliferation induced by IL-17 stimulation.