论文部分内容阅读
本文为研究Snail1基因对喉鳞癌细胞株Hep-2紧密连接蛋白表达及迁移能力的影响,将含有Snail1基因的短发夹RNA(Sh-RNA)的质粒转染喉鳞癌细胞株Hep-2,培养出可稳定传代的Snail1基因沉默的细胞(命名为Sh-snail1细胞)。课题组以蛋白免疫印迹技术检测紧密连接蛋白ZO-1、Occludin、Claudin-5的表达是否发生变化。然后设计伤口愈合实验检测其迁移能力,再以蛋白免疫印迹技术检测与细胞迁移能力密切相关的Rho GTP酶家族重要成员Rho A、Cdc42蛋白的表达变化。结果表明,Sh-snail1细胞紧密连接关键蛋白ZO-1、Occludin、Claudin-5表达明显上调,迁移能力明显减弱,且Rho A、Cdc42蛋白表达下调。该研究表明,喉鳞癌Hep-2细胞通过下调Snail1基因的表达使细胞间连接更加紧密,同时抑制Hep-2细胞的迁移能力,下调Rho A、Cdc42蛋白的表达。本文研究结果证实,Snail1基因与Hep-2细胞转移过程中的细胞间紧密连接打开以及细胞迁移能力增强密切相关,为靶向治疗喉鳞癌提供分子机制的研究依据。
In order to study the effect of Snail1 on the tight junction protein expression and migration ability of Hep-2 cells, the short hairpin RNA (Sh-RNA) containing Snail1 gene was transfected into Hep-2 cell line Hep-2 , A cell that silenced the passage of the Snail1 gene (named Sh-snail1 cells) was cultured. The research team detected the expression of tight junction proteins ZO-1, Occludin and Claudin-5 by Western blotting. The wound healing assay was then designed to detect the migration ability. The expression of Rho A and Cdc42, a key member of Rho GTPase family closely related to cell migration ability, was detected by Western blotting. The results showed that the expression of ZO-1, Occludin and Claudin-5 in Sh-snail1 cells was significantly up-regulated, the migration ability was significantly weakened, and the expressions of Rho A and Cdc42 were down-regulated. The study shows that Hep-2 cells down-regulate the expression of Snail1 gene to make cell junctions more tightly, inhibit Hep-2 cell migration and down-regulate Rho A and Cdc42 protein expression. The results of this study confirm that the Snail1 gene is closely related to the close intercellular junctional opening and cell migration in the process of Hep-2 cell metastasis, providing a basis for molecular mechanisms of targeted therapy of laryngeal squamous cell carcinoma.