过表达SOCS5对非小细胞肺癌增殖、迁移和侵袭的作用及机制研究

Effect and mechanism of overexpression of SOCS5 on the proliferation, migration and invasion of non-small cell lung cancer

目的分析细胞因子信号传导抑制因子5(SOCS5)对非小细胞肺癌(NSCLC)细胞增殖、迁移和侵袭能力的影响,并探讨其可能的作用机制。方法应用癌症基因组图谱(TCGA)数据库分析SOCS5在肺癌组织中的表达情况;western blot检测SOCS5在NSCLC细胞系PC-9、A549和SPC-A-1及正常支气管上皮细胞系16HBE中的表达水平;选择SOCS5表达水平最低的NSCLC细胞系进行SOCS5过表达质粒转染,实验设对照(NC)组和SOCS5过表达质粒转染(oe-SOCS5)组。采用MTS试验检测各组细胞增殖能力;划痕试验检测各组细胞迁移能力;Boyden试验检测各组细胞侵袭能力;western blot检测各组细胞中PI3K/Akt/mTOR信号通路活性。结果TCGA数据库分析结果显示,SOCS5 mRNA在肺腺癌和肺鳞癌组织中的表达水平显著低于正常肺组织(P<0.05)。western blot结果显示,SOCS5蛋白在PC-9、A549和SPC-A-1细胞中的表达水平均低于其在16HBE细胞中的表达水平(P<0.05);选择表达水平最低的PC-9细胞进行SOCS5过表达质粒转染,结果发现与NC组相比,oe-SOCS5组PC-9细胞增殖、迁移和侵袭能力均降低(P<0.05);western blot结果显示,与NC组相比,oe-SOCS5组细胞pPI3K、pAkt和pmTOR蛋白的表达水平均降低(P<0.05)。结论SOSC5在NSCLC中呈异常低表达,过表达SOCS5可抑制NSCLC细胞的增殖、迁移和侵袭能力,抑制PI3K/Akt/mTOR信号通路的激活可能是作用机制之一。

Objective To investigate the effects of suppressor of cytokine signaling 5(SOCS5)on the proliferation,migration and invasion of non-small cell lung cancer(NSCLC)cells and its possible mechanism.Methods The expression of SOCS5 in NSCLC tissues was analyzed by the Cancer Genome Atlas(TCGA)database.The expression levels of SOCS5 in NSCLC cell lines PC-9,A549,and SPC-A-1 cells and normal bronchial epithelial cell line 16HBE cells were detected by western blot.The NSCLC cell line with the lowest expression of SOCS5 was selected for the transfection of SOCS5 overexpression plasmids,and it was divided into the control(NC)group and the transfection group of SOCS5 overexpression plasmids(oe-SOCS5).The cell proliferation ability of each group was detected by the MTS test.The cell migration and invasion ability of each group were detected by the scratch test and Boyden test,respectively.The activity of cell PI3K/Akt/mTOR signaling pathway in each group was determined by western blot.Results The analysis results of TCGA database showed that the expression levels of SOCS5 mRNA in lung adenocarcinoma and squamous cell carcinoma tissues were significantly lower than that in normal lung tissues(P<0.05).The results of western blot showed that the expression levels of SOCS5 in PC-9,A549 and SPC-A-1 cells were significantly lower than that in 16HBE cells(P<0.05).The PC-9 cells with the lowest expression level of SOCS5 were transfected with SOCS5 overexpression plasmids,and the results showed that the proliferation,migration and invasion ability of PC-9 cells in the oe-SOCS5 group were significantly lower than those in the NC group(P<0.05).Moreover,compared with the NC group,the expression levels of pPI3K,pAkt and pmTOR proteins in the oe-SOCS5 group were decreased significantly(P<0.05).Conclusion There is low expression of SOCS5 in NSCLC cells.The overexpression of SOCS5 may inhibit the proliferation,migration and invasion of NSCLC cells,and the inhibition of PI3K/Akt/mTOR signaling pathway may be one of the mechanisms.