Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶8在肺腺癌中的作用及机制研究

The role of a disintegrin-like and metalloproteinase with throm bospondin typeⅠmotifs-8 in lung adenocarcinomaand its mechanism

目的探究Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶8(ADAMTS8)基因在肺腺癌中的作用及机制。方法选取肺腺癌数据集GSE75037和GSE116959以及TCGA数据库中肺腺癌转录组数据和临床信息;使用R软件分析肺腺癌组织中差异基因,并对差异基因进行基因本体论(GO)富集分析和京都基因和基因组百科全书(KEGG)信号通路富集分析;分析ADAMTS8基因在肺腺癌中的表达及与患者生存和预后的相关性;CCK-8实验检测ADAMTS8过表达或敲低对A549细胞增殖的影响;Western blot检测ADAMTS8过表达或敲低对Wnt/β-catenin信号通路的影响。结果联合分析发现,有395个基因在肺腺癌样本中发生上调,716个基因发生下调;ADAMTS8在肺腺癌组织中低表达,其表达水平与肿瘤分期有关;ADAMTS8低表达与患者生存时间短、预后差相关,Cox回归分析表明,ADAMTS8可作为肺腺癌患者独立的预后标志物;CCK-8结果表明,过表达ADAMTS8能抑制A549细胞的增殖,敲低ADAMTS8能促进A549细胞的增殖(P<0.05);Western blot结果表明,过表达ADAMTS8后,细胞中β-catenin、cyclin D1和c-Myc的蛋白水平显著降低(P<0.05),而敲低ADAMTS8可引起细胞中β-catenin、cyclin D1和c-Myc的蛋白水平显著升高(P<0.05)。结论 ADAMTS8在肺腺癌中低表达,可作为患者独立的预后标志物,过表达ADAMTS8可能是通过抑制Wnt/β-catenin信号通路活性发挥对肺腺癌细胞增殖的抑制作用。

Objective To explore the role of a disintegrin-like and metalloproteinase with throm bospondin typeⅠmotifs-8(ADAMTS8)gene in lung adenocarcinoma and its mechanism.Methods Selected lung adenocarcinoma data sets GSE75037 and GSE116959 and lung adenocarcinoma transcriptome data and clinical information from the TCGA database;useed R software to analyze differential genes in lung adenocarcinoma tissues,and performed GO enrichment analysis and KEGG signal pathway enrichment for differential genes set analysis;analyzed the expression of ADAMTS8 gene in lung adenocarcinoma and its correlation with patient survival and prognosis;CCK-8 experiment detects the effect of ADAMTS8 overexpression or knockdown on the proliferation of A549 cells;Western blot detected ADAMTS8 overexpression or knockdown effect on Wnt/β-catenin signaling pathway.Results The combined analysis found that 395 genes were up-regulated in lung adenocarcinoma samples,and 716 genes were down-regulated;ADAMTS8 was under-expressed in lung adenocarcinoma tissues,and its expression level was related to tumor stage;low-expression of ADAMTS8 was associated with shorter survival time,poor prognosis,Cox regression analysis showed that ADAMTS8 can be used as an independent prognostic marker for patients with lung adenocarcinoma;CCK-8 results showed that overexpression of ADAMTS8 can inhibit the proliferation of A549 cells,knocking down ADAMTS8 can promote the proliferation of A549 cells(P<0.05);Western blot results showed that after overexpression of ADAMTS8,the protein levels ofβ-catenin,cyclin D1 and c-Myc in cells were significantly reduced(P<0.05),while knocking down ADAMTS8 could cause the protein levels ofβ-catenin,cyclin D1 and c-Myc in cells increased significantly(P<0.05).Conclusions ADAMTS8 is under-expressed in lung adenocarcinoma and can be used as an independent prognostic marker for patients.Overexpression of ADAMTS8 may inhibit the proliferation of lung adenocarcinoma cells by inhibiting the activity of Wnt/β-catenin signaling pathway.