SOX30基因调控桥粒基因DSC3抑制肺腺癌的增殖与迁移

Inhibition of proliferation and migration of lung adenocarcinoma cells by SOX30 gene via regulation of DSC3 gene

目的:探讨SOX30基因对桥粒基因DSC3的表达调控及其在肺腺癌发生中的作用。方法:利用基因功能富集分析(GO分析)获取SOX30调控的重要基因及通路;利用JASPAR数据库预测DSC3启动子区SOX30蛋白结合位点;利用TCGA数据库分析SOX30与DSC3在肺癌中表达的相关性,并用逆转录PCR检测A549细胞高表达SOX30后对DSC3 mRNA表达的影响;同时检测SOX30基因敲除小鼠肺组织中DSC3 mRNA的表达情况;利用平板细胞集落形成实验及细胞划痕愈合实验分析DSC3是否参与SOX30对肺腺癌细胞的增殖与迁移抑制。结果:GO分析显示SOX30基因与黏着斑、锚定连接、黏着连接等细胞连接基因表达显著相关;DSC3启动子区存在多个SOX30蛋白结合位点;SOX30与DSC3在肺腺癌中表达正相关(r=0.154,P=0.000);高表达SOX30后,A549细胞中DSC3 mRNA水平较对照组显著上调(P<0.05);敲除SOX30基因后,纯合子(SOX30-/-)小鼠肺组织中DSC3 mRNA水平显著低于杂合子(SOX30+/-)及野生型(SOX30+/+)小鼠(P均<0.01);在高表达SOX30后的A549细胞中,干扰DSC3的表达后将显著减弱SOX30对A549细胞的增殖和迁移抑制(P均<0.05)。结论:SOX30是调控DSC3表达的关键分子,DSC3是SOX30发挥抑癌功能的重要下游基因,SOX30-DSC3调控可能在肺腺癌发生发展中起重要作用。

OBJECTIVE：To investigate interactions between SOX30 and DSC3 genes and their roles in the development of lung adenocarcinoma.METHODS：The GO enrichment analysis was used to screen possible downstream genes and pathways of SOX30 transcriptional regulation.The JASPAR database was used to predict the SOX30 transcription binding sites in the promoter region of DSC3.Expression data of SOX30 and DSC3 were downloaded from the TCGA database,and their relationships were evaluated using the Spearman rank correlation analysis.The reverse transcription-PCR was used to measure the mRNA levels of DSC3 in A549-SOX30＋DSC3 miRNA stably transfected cells and the lung tissues of SOX30＋/＋,SOX30＋/-and SOX30-/-mice.The colony formation and wound healing assays were used to confirm whether the inhibition of the proliferation and migration of SOX30 in A549 cells depended on the expression of DSC3 or not.RESULTS：The GO enrichment analysis shows the function of SOX30 gene was significantly related to the cell junction proteins,such as focal adhesion,anchoring junction,and adhesion junction.And several SOX30 binding sites were found in the promoter region of DSC3.Furthermore,expression of DSC3 was in positive correlation with that of SOX30 in human lung adenocarcinoma（r=0.154,P=0.000）.In addition,the ectopic expression of SOX30 in A549 cell lines dramatically increased DSC3 mRNA level （P〈0.05）,and knockout of SOX30 significantly decreased DSC3 mRNA level in lung tissues of SOX30-/-mice （P〈0.05）.In addition,knockdown of DSC3 attenuated the inhibition effects of SOX30 on proliferation and migration in A549 cells （P〈0.05）.CONCLUSION：SOX30 was a critical transcription factor in the expression regulation of DSC3,and DSC3 was an important downstream gene of SOX30.Therefore,SOX30-DSC3 may play an important role in the development of lung adenocarcinoma.