长链非编码RNA-LINC00485在肺癌中的表达及对细胞增殖和迁移的影响  

Expression of long-chain non-coding RNA-LINC00485 in lung cancer and its effect on proliferation and migration of lung cancer cells

作　　者：王向辉[1] 黄江平[1] 崔丰和[1] 钱海云[1] WANG Xiang-hui;HUANG Jiang-ping;CUI Feng-he;QIAN Hai-yun(Department of Cardiothoracic Surgery,Jingzhou Hospital,Tongji Medical College,Huazhong University of Science and Technology,Jingzhou434020,China)

机构地区：[1]华中科技大学同济医学院附属荆州医院心胸大血管外科,荆州434020

出　　处：《临床与实验病理学杂志》2019年第1期42-46,共5页Chinese Journal of Clinical and Experimental Pathology

摘　　要：目的观察长链非编码RNA-LINC00485(LINC00485)在肺癌细胞株及组织中的表达,探讨其对肺癌细胞增殖和迁移的影响及其作用机制。方法采用qRT-PCR检测LINC00485在4种肺癌细胞株(H1975、A549、HCC827、H1299)、正常肺泡上皮细胞HPAEPIC和12例肺癌组织及癌旁组织中的差异性表达。通过生物信息学方法预测LINC00485可能结合的微小RNA(miRNA)及靶基因,将靶向沉默LINC00485的小干扰RNA(siRNA)通过脂质体转染至HCC827细胞。应用qRT-PCR检测LINC00485、miR-361-5p、p21活化蛋白激酶2(PAK2) mRNA的表达水平。采用Western blot法检测PAK2蛋白的表达水平。应用MTS法检测细胞增殖能力、细胞划痕实验检测细胞迁移能力。结果与正常肺泡上皮相比,LINC00485在肺癌细胞株中均呈高表达(P <0. 05),其中在HCC827细胞中表达水平最高。LINC00485在肺癌组织中的表达水平高于其癌旁组织(P <0. 01)。下调HCC827细胞LINC00485的表达后,miR-361-5p的表达上调(P <0. 01),PAK2 mRNA和蛋白的表达下调(P <0. 01),HCC827细胞增殖能力降低(P <0. 05),细胞迁移能力下降(P <0. 01)。结论 LINC00485在肺癌细胞株和组织中表达升高,下调LINC00485可通过调控miR-361-5p和PAK2基因的表达,抑制肺癌HCC827细胞的增殖和迁移能力。Purpose To observe the expression of long-chain non-coding RNA-LINC00485(LINC00485)in lung cancer cell lines and tissues,and to investigate its effect on the proliferation and migration of lung cancer cells and its mechanism.Methods Quantitative real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect differential expression of LINC00485 in four lung cancer cell lines(H1975,A549,HCC827,H1299),normal alveolar epithelial cells HPAEPIC,and in 12 cases lung cancer tissues and adjacent tissues.Bioinformatics methods were used to predict the microRNA(miRNA)that LINC00485 may bind and target gene that miRNA may bind.Small interfering RNAs(siRNAs)that target silencing LINC00485 were transfected into HCC827 cells by liposomes.The expression levels of LINK00485,miR-361-5p,and p21 activated protein kinase 2(PAK2)mRNA were detected by qRT-PCR.The expression level of PAK2 protein was detected by Western blot.The cell proliferation ability was measured by MTS assay.Cell scratch assay was used to detect cell migration.Results Compared with normal alveolar epithelium,LINC00485 was highly expressed in lung cancer cell lines(P<0.05),and the expression level was highest in HCC827 cells.The expression of LINC00485 in lung cancer tissues was higher than that in adjacent tissues(P<0.01).After down-regulation of LINC00485 expression in HCC827 cells,the expression of miR-361-5p was up-regulated(P<0.01),the expression of PAK2 mRNA and protein was down-regulated(P<0.01),the proliferative capacity of HCC827 cells was decreased(P<0.05),and the ability of cell migration was decreased(P<0.01).Conclusion The expression of LINC00485 is increased in lung cancer cell lines and tissues.Down-regulation of LINC00485 can inhibit the proliferation and migration of lung cancer HCC827 cells by regulating the expression of miR-361-5p and PAK2 genes.

关 键 词：肺肿瘤 长链非编码RNA-LINC00485 增殖 迁移 QRT-PCR 

分 类 号：R734.2[医药卫生—肿瘤]