SATB1在人喉鳞状细胞癌细胞株Hep-2侵袭转移中的作用及其机制研究

Role of SATB1 in invasion and metastasis of human laryngeal squamous cell carcinoma cell line Hep-2 and its mechanisms

目的探讨富集AT序列的特异性结合蛋白1(SATB1)在人喉鳞状细胞癌细胞株Hep-2侵袭转移中的作用及其机制。方法体外培养Hep-2细胞并分为两组,实验组细胞转染靶向SATB1基因的小干扰RNA(SATB1-siRNA),对照组细胞转染阴性对照siRNA(NC-siRNA)。分别于转染前和转染后48 h,采用实时荧光定量PCR检测两组细胞中SATB1、E-钙黏素(E-cadherin)和锌指转录因子(Snail)基因的mRNA表达水平,Transwell小室侵袭实验检测两组细胞的侵袭能力,细胞划痕实验检测两组细胞划痕后48 h的转移能力。结果转染前两组细胞中SATB1、E-cadherin和Snail基因的mRNA相对表达量比较,差异均无统计学意义(均P>0.05)。与对照组相比,实验组转染后细胞中SATB1和Snail基因的mRNA相对表达量均降低,而E-cadherin基因的mRNA相对表达量升高,差异均具有统计学意义(均P<0.05)。转染后两组细胞的Transwell穿膜细胞数和细胞迁移距离均减少,且实验组低于对照组,差异均具有统计学意义(均P<0.05)。结论SATB1可促进Hep-2细胞的侵袭转移,其作用机制可能为下调E-cadherin和上调Snail基因表达从而诱导上皮间质转化。

Objective To investigate the role of special AT-rich sequence-binding protein 1(SATB1)in invasion and metastasis of human laryngeal squamous cell carcinoma cell line Hep-2 and its mechanism.Methods Hep-2 cells were cultured in vitro.The cell lines were divided into two groups.The cells in the experimental group was transfected with Lipofectamine 2000 liposome for SATB1 small interfering RNA(SATB1-siRNA),and the cells in the control group was transfected with Lipofectamine 2000 liposome for NC-siRNA.The before and after transfection SATB1,E-cadherin and Snail genes relative expression levels between the two groups were detected and compared using RT-PCR.Transwell cell invasion experiments were applied to evaluate the cells invasive ability of the two groups.The scratch experiments were applied to evaluate the 48 h cell migration ability of the two groups.Results The differences in the relative expression level of SATB1,E-cadherin and Snail genes in pre-transfected laryngeal squamous cell line Hep-2 between the two groups were not statistically significant(all P>0.05).Compared with the control group,the relative expression levels of the mRNA of SATB1 and Snail genes decreased in the experimental group after transfection,while the relative expression levels of the mRNA of E-cadherin gene increased,and the differences were statistically significant(all P<0.05).After the transfection,the number of transwell membrane cells and cell migration distances of the two groups of cells decreased.The values of cell number and migration distance in the experimental group were lower than those in the control group,and the differences were statistically significant(all P<0.05).Conclusions SATB1 can promote the invasion and metastasis of Hep-2 cells,and its mechanism may be related to the down-regulation of E-cadherin and the up-regulation of Snail gene expression to induce epithelial-mesenchymal transition.