siRNA下调Gli基因表达对肺鳞癌SK-MES-1细胞生物学行为的影响-东南大学学报医学版

siRNA下调Gli基因表达对肺鳞癌SK-MES-1细胞生物学行为的影响

单位：1. 河北医科大学第四医院胸二科, 河北石家庄 050011;
2. 河北医科大学期刊社, 河北石家庄 050017

分类号：R734.2

出版年·卷·期（页码）：2016·35·第六期（908-913）

摘要：

目的：探讨Gli表达下调对肺鳞癌SK-MES-1细胞周期分布和上皮-间质转化能力的影响及其分子机制。方法：将Gli1和Gli2 siRNA与空白对照siRNA分别转染SK-MES-1细胞48 h，实时荧光定量PCR法检测Gli1、Gli2 mRNA表达水平；Western blot法检测SK-MES-1细胞Gli1、Gli2、Cyclin D1、Cyclin E、E-cadherin、N-cadherin蛋白的表达；流式细胞术检测SK-MES-1细胞周期分布；Transwell侵袭实验检测细胞侵袭能力。结果：与空白对照siRNA组相比，Gli1和Gli2 siRNA可明显抑制SK-MES-1细胞Gli1、Gli2 mRNA及蛋白表达，下调Cyclin D1、Cyclin E、N-cadherin蛋白的表达，增加E-cadherin蛋白表达，将细胞周期明显阻滞在G0/G1期并降低细胞的侵袭能力，差异均有统计学意义（P<0.01）。结论：Gli蛋白表达可能在肺鳞癌的发生发展中具有重要作用，Gli1和Gli2表达下调可导致肺鳞癌细胞周期分布和上皮-间质转化能力的改变，这可能与Cyclin D1、Cyclin E、E-cadherin、N-cadherin蛋白变化相关。

Objective: To detect the expression of Gli in lung squamous carcinoma, and to analyze the effects of down-regulation of Gli expression on malignant biological behavior of lung squamous carcinoma SK-MES-1 cells and to explore their molecular mechanisms. Methods: Gli1, Gli2 siRNA and control siRNA were transfected into SK-MES-1 cells for 48 h respectively. The Gli-1, Gli-2 mRNA expression were detected by real time fluorescence quantitative PCR method. The protein expression of Gli-1, Gli-2, Cyclin D1, CyclinE, N-cadherin and E-cadherin were detected by Western blot assay. The cell cycle were observed by flow cytometry. Transwell assay was performed to detect cell invasion ability. Results: Compared with Control siRNA, Gli1 & Gli2 siRNA could significantly inhibit the Gli-1, Gli-2 mRNA and protein expression in SK-MES-1 cells, which at last downregulated the CyclinD1, CyclinE, N-cadherin protein expression, and enhanced E-cadherin expression. Downregulation of Gli-1, Gli-2 could arrest cell cycle at G0/G1 phase and decrease cell invasion ability(P<0.01). Conclusion: Gli may play an important role in the development of lung squamous carcinoma. The downregulation of Gli expression can lead to changes in the cell cycle and invasion of lung squamous carcinoma, which may be associated with Cyclin D1, Cyclin E, N-cadherin and E-cadherin proteins.

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