FoxO3a对胆管癌细胞增殖及凋亡的影响

Effect of FoxO3a on proliferation and apoptosis of cholangiocarcinoma cells

目的研究叉头框转录因子O3a(FoxO3a)对胆管癌细胞增殖及凋亡的影响。方法胆管癌细胞RBE中转染FoxO3a过表达载体,qRT-PCR和Western blotting检测细胞中FoxO3a mRNA和蛋白水平,MTT检测细胞增殖,流式细胞术检测细胞凋亡水平,Western blotting检测细胞中β-连环蛋白(β-catenin)、c-Myc、细胞周期蛋白D1(cyclin D1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)蛋白水平。结果转染FoxO3a过表达载体后,RBE细胞中FoxO3a表达水平明显高于未转染的RBE细胞(P<0.05),而转染对照载体的RBE细胞中FoxO3a表达水平与未转染的细胞相比,差异无统计学意义(P>0.05)。转染FoxO3a过表达载体后RBE细胞A值从0.73±0.12降低至0.52±0.04,细胞凋亡率从(8.36±0.68)%升高至(25.62±2.52)%,细胞中Cleaved Caspase-3水平也升高,β-catenin、c-Myc、cyclin D1蛋白水平降低,与未转染的细胞相比,差异有统计学意义(P<0.05)。结论 FoxO3a表达上调后可以抑制胆管癌细胞增殖,诱导细胞凋亡,增加Cleaved Caspase-3的活化水平,抑制Wnt信号通路激活。

Objective To investigate the effect of FoxO3a on proliferation and apoptosis of cholangiocarcinoma cells.Methods FoxO3a over expression vector was transfected into cholangiocarcinoma cell line RBE,the levels of FoxO3a mRNA and protein in cells were detected by qRT-PCR and Western blotting. Cell proliferation was detected by MTT.Cell apoptosis was detected by flow cytometry,Western blotting was used to detect the levels of β-catenin,c-Myc,cyclin D1 and Cleaved Caspase-3 proteins in the cells. Results After transfection of FoxO3a over expression vector,the expression of FoxO3a in RBE cells was significantly higher than that in RBE cells without transfection（P〈0. 05）,the expression of FoxO3a in RBE cells transfected with empty vector showed no significant change compared with that of cells without transfection（P〈0. 05）. After transfection of FoxO3a over expression vector,the A value of RBE cells was decreased from 0. 73 ± 0. 12 to 0. 52 ± 0. 04,the apoptosis rate was increased from（8. 36 ± 0. 68） % to（25. 62 ±2. 52） %,the level of Cleaved Caspase-3 was also elevated in the cells,the levels of β-catenin,c-Myc and cyclin D1 proteins were decreased,the difference was statistically significant between the transfected cells and the non transfected cells（P〈0. 05）. Conclusion Up regulation of FoxO3a expression can inhibit proliferation of cholangiocarcinoma cells,apoptosis induction,increase the activation level of Caspase-3,inhibition of activation of Wnt signaling pathway.