BNC1 促进肺鳞状细胞癌细胞迁移和侵袭

BNC1 promotes cell migration and invasion of lung squamous cell carcinoma

目的探索碱性核蛋白1(basonuclin 1,BNC1)基因对肺鳞状细胞癌细胞迁移和侵袭能力的影响及其作用机制。方法利用生物信息学分析在线数据库中BNC1基因在肺鳞状细胞癌患者中的表达规律及对生存的影响;采用免疫组织化学对组织芯片进行染色并分析BNC1的蛋白水平;在癌症细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库中筛选BNC1高表达和低表达的肺鳞状细胞癌细胞系;对敲低和过表达的肺鳞状细胞癌细胞H226、SK-MES1进行划痕愈合实验、Transwell迁移和侵袭实验,检测细胞的迁移与侵袭能力的变化;采用RT-qPCR验证10个文献来源的有关迁移与侵袭高度相关的基因;采用Western blot对筛选的调控基因进一步验证。结果生物信息学分析发现BNC1高表达于肺鳞状细胞癌组织且与较低生存率相关(P=0.047);肺鳞状细胞癌组织中BNC1的表达高于癌旁组织(P<0.05);H226细胞系BNC1表达较高,而SK-MES1细胞系BNC1表达较低;敲低BNC1能够抑制肺鳞状细胞癌细胞的迁移和侵袭(P<0.05),过表达BNC1可增强肺鳞状细胞癌细胞的迁移和侵袭(P<0.05);在mRNA层面筛选BNC1下游调控分子,发现E-钙粘蛋白(E-cadherin,CDH1)在BNC1过表达时减低(P<0.05),在BNC1敲低时升高(P<0.05),Western blot验证结果与mRNA表达一致(P<0.05)。结论BNC1可能是肺鳞状细胞癌患者的预后不良基因,BNC1基因可能通过下调CDH1的表达,促进肺鳞状细胞癌细胞的迁移和侵袭。

Objective To explore the mechanism and effects of Basonuclin 1(BNC1)on the migration and invasion abilities of lung squamous cell carcinoma.Methods Bioinformatics technology was used to analyze the expression patterns and survival impact of BNC1 in patients with lung squamous cell carcinoma in online databases;Tissue microarrays were stained by immunohistochemistry assay and analyzed for BNC1 protein level.Cell lines for lung squamous cell carcinoma with high and low BNC1 expression were screened in the Cancer Cell Line Encyclopedia(CCLE)database.Wound healing experiment and transwell assay were performed on lung squamous cell carcinoma cell models with knockdown and overexpression of BNC1,and the migration and invasion rates of each cell line were detected;Quantitative reverse transcription PCR(RT-qPCR)was used to validate the migration and invasion of 10 highly related genes derived from literature.Western blotting was used to validate the selected regulatory genes.Results Bioinformatics technology found that BNC1 was highly expressed in lung squamous cell carcinoma tissues and was associated with lower survival rate(P=0.047).The expression of BNC1 in lung squamous cell carcinoma was higher than that in paracancerous tissues(P<0.05).BNC1 had higher expression in the H226 cells and lower expression in the SK-MES1 cells.BNC1 knockdown inhibited the migration and invasion abilities of lung squamous cell carcinoma cells(P<0.05),and BNC1 overexpression enhanced the migration and invasion abilities of lung squamous cell carcinoma cells(P<0.05).Screening of BNC1 downstream regulatory molecules at the mRNA level showed that CDH1 was decreased when BNC1 was overexpressed(P<0.05),and increased when BNC1 was knocked down(P<0.05).The results of mRNA level were consistent with the results verified by Western blotting(P<0.05).Conclusion BNC1 may be a gene predicting poor prognosis for lung squamous cell carcinoma patients,and BNC1 may likely to promote cell migration and invasion of lung squamous cell carcinoma by downregulating CDH1 expression.