siRNA下调Gli基因影响食管腺癌OE33细胞生长转移的实验研究

Silencing Gli gene with si RNA modulates growth and invasion of esophageal adenocarcinoma OE33 cells

目的研究Gli表达下调对食管腺癌OE33细胞生长转移的影响,并探讨其相关的分子机制。方法将Gli1、Gli2 si RNA和空白对照si RNA转染OE33细胞48 h,实时荧光定量聚合酶链反应检测OE33细胞Gli1和Gli2 m RNA表达水平;采用Western blot检测OE33细胞中Gli1、Gli2、Cyclin D1、p27、E-cadherin及N-cadherin蛋白的表达水平;流式细胞术检测OE33细胞增殖和细胞周期分布;Transwell实验检测细胞侵袭能力。结果采用Gli1和Gli2 si RNA抑制Gli1和Gli2 m RNA及蛋白表达后,可下调Cyclin D1和N-cadherin蛋白表达,增加E-cadherin和p27蛋白表达,同时抑制细胞增殖,将细胞周期阻滞在G_0/G_1期并降低细胞的侵袭能力,与对照组比较,差异有统计学意义(P<0.05)。结论 Gli在食管腺癌的生长转移中具有重要作用,敲掉Gli1和Gli2基因表达可抑制食管腺癌细胞周期分布和上皮-间质转化能力,这可能与Cyclin D1、p27、E-cadherin及N-cadherin蛋白变化相关。

To explore the effect of down-regulation of Gli expression on growth and invasion of esophageal adenocarcinoma OE33cells and to discuss the molecular mechanisms.Methods Gli1and Gli2siRNA and control siRNA were respectively transfected into OE33cells for48h.Expressions of the Gli1and Gli2mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot respectively.The expressions of cyclin D1,p27,E-cadherin and N-cadherin proteins were observed by Western blot.The cell cycle was determined by flow cytometry.In addition,invasion ability was detected by Transwell assay.Results After transfection of Gli1and Gli2siRNA,Gli1and Gli2expressions were inhibited,the expressions of cyclin D1and N-cadherin proteins were downregulated,and E-cadherin and p27expressions were increased(P<0.05).After Gli1and Gli2were knocked down,cell cycle was arrested at G0/G1phase and cell invasion ability was decreased compared with the control siRNA(P<0.05).Conclusions Gli may play an important role in the growth and invasion of esophageal adenocarcinoma.Gli1and Gli2knockout can inhibit the cell cycle and epithelial-mesenchymal transition,which may be associated with cyclin D1,p27,E-cadherin and N-cadherin.