SOX9 drives the epithelial-mesenchymal transition in non-small-cell lung cancer through the Wnt/-catenin pathway
SOX9 drives the epithelial-mesenchymal transition in non-small-cell lung cancer through the Wnt/-catenin pathway
复制标题
SOX9 通过 Wnt/β-连环蛋白途径驱动非小细胞肺癌的上皮间质转化
DOI：
10.1186/s12967-019-1895-2
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发表时间：
2019-05-06
影响因子：
7.4
通讯作者：
Gong, Li-Yun
中科院分区：
文献类型：
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作者：
Huang, Jing-Qiang;Wei, Fa-Kai;Gong, Li-Yun
背景：在非小细胞肺癌（NSCLC）中，癌细胞的远处转移是导致肿瘤死亡和预后不良的危险因素。SOX9表达升高与NSCLC的临床分期和预后不良有关，但SOX9促进NSCLC转移的分子机制尚不清楚。方法采用(2)试验和Spearman分析法，对142例非小细胞肺癌免疫组化诊断标本进行SOX9表达与T、N、M分级的关系分析。我们还通过慢病毒构建体转染获得了sox9过表达和sox9敲低细胞系及其相应的对照细胞系。在体内实验中，将sox9过表达和sox9敲低的NSCLC细胞注射到斑马鱼体内，观察其远处转移。采用基因集富集分析（GSEA）分析SOX9过表达与Wnt/-catenin通路的相关性。采用荧光素酶法检测TCF/LEF的转录活性，采用western blot和免疫荧光法检测-catenin在sox9过表达、sox9敲低及其对照细胞系中的易位。结果在142例非小细胞肺癌免疫组化诊断标本中，SOX9过表达与T、N、M分期相关（p分别为0.03、0.000、0.032）。SOX9过表达可降低上皮细胞标志物E-cadherin和-catenin的表达，增加间质细胞标志物N-cadherin和vimentin的表达。体内实验显示sox9过表达细胞有远处转移，而在sox9敲低细胞中没有观察到这种转移。这些发现表明SOX9通过促进NSCLC细胞的EMT来促进远处转移。GSEA显示SOX9过表达与Wnt/-catenin通路显著相关，该通路及其下游靶基因中emt相关蛋白的表达证实了这一点。SOX9过表达还可以增强TCF/LEF的转录活性，促进-catenin的核易位，增加GSK3 Ser9位点的磷酸化。此外，抑制-catenin抑制SOX9过表达促进转移的作用。本研究首次报道了SOX9与临床TNM分期相关，并表明SOX9通过Wnt/-catenin通路促进迁移、侵袭和EMT过程。
BackgroundThe distant metastasis of cancer cells is a risk factor for tumor lethality and poor prognosis in non-small-cell lung carcinoma (NSCLC). Increased SOX9 expression has been associated with clinical stage and poor prognosis in NSCLC, but the molecular mechanisms by which SOX9 promotes metastasis in NSCLC are still unknown.MethodsThe relationship between SOX9 expression and T, N, M classification was assessed using the (2) test and Spearman's analysis in 142 immunohistochemically diagnosed specimens of NSCLC. We also generated SOX9-overexpression and SOX9-knockdown cells lines and their corresponding control cell lines by transfection with lentiviral constructs. In vivo assay, SOX9-overexpressing and SOX9-knockdown NSCLC cells were injected in zebrafish to examine distance metastasis. Gene set enrichment analysis (GSEA) was applied to analysis the correlation between SOX9 overexpression and Wnt/-catenin pathway. Luciferase assay was used to check transcriptional activity of TCF/LEF and western blot and immunofluorescence was employed to detect -catenin translocation in SOX9-overexpression, SOX9-knockdown and their corresponding control cell lines.ResultsWe found that SOX9 overexpression correlates with the T, N and M stage significantly (p=0.03, 0.000, and 0.032 respectively) in 142 immunohistochemically diagnosed specimens of NSCLC. SOX9 overexpression was found to decrease the expression of the epithelial cell markers E-cadherin and -catenin and increase the expression of the mesenchymal cell markers N-cadherin and vimentin. An in vivo assay showed distant metastasis of the SOX9-overexpressing cells, which was not observed in the SOX9-knockdown cells. These findings indicate that SOX9 promotes distant metastasis by promoting EMT in NSCLC cells. GSEA showed that SOX9 overexpression was significantly correlated with the Wnt/-catenin pathway which was corroborated by the expression of EMT-associated proteins in this pathway and its downstream target genes. SOX9 overexpression was also found to enhance the transcriptional activity of TCF/LEF, promote the nuclear translocation of -catenin and increase the phosphorylation of GSK3 at Ser9. Further, inhibition of -catenin suppressed the metastasis-promoting effects of SOX9 overexpression.ConclusionsThis study is the first to report that SOX9 is associated with clinical TNM stage and indicates that SOX9 promotes migration, invasion and the EMT process through the Wnt/-catenin pathway.