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目的观察bcl-2、bax mRNA在三氧化二砷(As_2O_3)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；逆转录一聚合酶链反应(RT- PCR)检测bcl-2、bax mRNA的表达。结果As_2O_3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；24 h实验组为0．01±0．01、0．43±0．02。hax/GAPDH mRNA A比值QBC939、RBE细胞对照组分别为0．21±0．01、0．42±0．04；24 h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As_2O_3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 observe the expression of bcl-2 and bax mRNA in human cholangiocarcinoma cell line QBC939 and RBE induced by As 2 O 3. Methods Rhodamine123 staining was used to detect the changes of mitochondrial membrane permeability. The expressions of bcl-2 and bax mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results Compared with control group, the intensity of Rhodamine123 fluorescence staining in experimental group was significantly weaker than that in control group. The ratio of bcl-2 / β-actin mRNA A was detected by gel imaging. The control group of QBC939 and RBE cells were 0.41 ± 0.03 and 0.84 ± 0.03; 24 h experimental group was 0.01 ± 0.01,0.43 ± 0.02. The ratios of hax / GAPDH mRNA A in QBC939 and RBE cells were 0.21 ± 0.01 and 0.42 ± 0.04, respectively, and those in 24 h group were 1.44 ± 0.16 and 1.15 ± 0.21 , The difference between the control group and the experimental group was statistically significant (P <0.01). Conclusion As 2 O 3 may increase the permeability of mitochondrial membrane, decrease the expression of bcl-2 mRNA and up-regulate the expression of bax mRNA in cholangiocarcinoma cell line, which may activate the mitochondrial apoptotic signal transduction pathway.