目的观察bcl-2、bax mRNA在三氧化二砷(As2O3)诱导人类胆管癌株QBC939、RBE细胞凋亡中的表达。方法Rhodamine123染色检测线粒体膜通透性变化;逆转录-聚合酶链反应(RT-P(浆)检测bcl-2、bax mRNA的表达。结果As2O3干预,实验组Rhodamine123荧光染色强度较对照组明显减弱;凝胶成像检测bcl-2/β-actin mRNA A比值,QBC939、RBE细胞对照组分别为0.41±0.03、0.84±0.03;24h实验组为0.01±0.01、0.43±0.02。bax/GAPDH mRNA A比值QBC939、RBE细胞对照组分别为0.21±0.01、0.42±0.04;24h实验组为1.44±0.16、1.15±0.21,对照组与实验组比较差异均有统计学意义(P〈0.01)。结论As2O3可能增加胆管癌细胞株线粒体膜的通透性,下调bcl-2 mRNA表达。上调bax mRNA的表达,可能启动了线粒体凋亡信号传导途径。
Objective To research the expression of bcl-2/bax mRNA in As2O3 induced cholan- giocarcinoma cells line ( QBC939, RBE) apoptosis in vitro. Methods The mitochondria permeability was measured by Rhodamine123 assay; The bcl-2/bax mRNA expression was detected by RT-PCR. Results The Rhodamine123 fluorescein stain intensity in the experiment group after treatment with As2O3 was decreased as compared with that in the control group. Gel imaging system revealed that the bcl-2/β-actin mRNA optical density ratio of QBC939/RBE cells in control group was 0.41 ± 0.03/0.84 ± 0.03 and 0.01 ± 0.01/0.43 ± 0.02 in experiment group respectively, and the bax/GAPDH mRNA optical density ratio of QBC939/RBE cells in control group was 0.21 ± 0.01/0.42 ± 0.04 and 1.44 ± 0.16/1.15 ± 0.21 in experiment group respectively. There was significant difference between control group and experiment group (P 〈 0.01). Conclusion As2O3 may induce mitochondrial membrane depolarization, downregulate the expression of bcl-2 mRNA and up-regulate the expression of bax mRNA expression in cholangiocarcinoma cells line. As2O3 may activate the mitochondria apoptosis signal transduction pathway.