Survivin反义寡核苷酸对肝门部胆管癌细胞株FRH-0201作用的研究

Effect of antisense oligodeoxynucleotide targeting survivin on hilar cholangiocarcinoma cell line FRH-0201

目的探讨survivin反义寡核苷酸抑制survivin基因表达对肝门部胆管癌细胞FRH-0201的作用。方法脂质体介导survivin ASODN转染肝门部胆管癌细胞FRH-0201；倒置相差显微镜观察细胞形态学变化；RT—PCR、Western印迹法检测survivin mRNA及蛋白表达水平；流式细胞仪检测细胞凋亡率。结果survivin反义寡核苷酸作用后肝门部胆管癌细胞FRH-0201 survivin mRNA及蛋白表达水平较未转染组显著下降（mRNA：0．51±0．03vs0．82±0．02，P〈0．05；蛋白：1．82±0．16vs3．08±0．27．P〈0．05）．细胞出现凋亡形态学变化，细胞凋亡率增高（11．50±1．49％vs0．39±0．08％，P〈0．05）。结论survivin反义寡核苷酸能有效下调survivin基因表达，诱导肝门部胆管癌细胞凋亡。

Objective To investigate the effect of antisense oligodeoxynucleotide targeting sur vivin （survivin ASODN） on hilar cholangioeareinoma cell line FRH-0201 depressing the expression of survivin. Methods Survivin ASODN was transfected into hilar cholangiocarcinoma cell line FRH 0201 by liposome. Morphological changes were observed by inverted phase contrast microscope. RT-PCR and Western blot methods were performed to detect the expressions of survivin mRNA and protein re spectively. The changes in cell apoptosis were detected by flow cytometry. Results The expression of survivin mRNA and protein was significantly decreased in survivin ASODN group than that in the con trolgroup（mRNA： 0.51±0.03 vs 0.82±0.02,P〈0.05;protein： 1.82±0.16 vs 3.08±0.27, P〈 0.05）. The morphological apoptotic changes were observed and the apoptosis rate was increased （11.50±1.49% vs 0. 39±0. 08%, P〈0.05）. Conclusion Survivin ASODN can induce hilar cholangiocarcinoma cell line FRH-0201 into apoptosis by decreasing the expression of survivin.