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背景与目的:胆囊癌由于缺乏特异性的临床表现,确诊时多属晚期,手术切除率较低;且常规化疗药物对胆囊癌的疗效较差,因此需寻找治疗胆囊癌新的有效药物。三氧化二砷在治疗急性早幼粒细胞白血病中取得了突破性成果,近年来它在实体瘤的研究中颇受关注。本研究旨在探讨As2O3对人胆囊癌GBC细胞系诱导凋亡的生物学效应及作用机制。方法:用Hoechest33258染色、流式细胞仪和DNA凝胶电泳研究As2O3诱导细胞凋亡情况。用W estern b lot分析As2O3对凋亡相关蛋白Bc l-2、Bc l-xL、B id、Bak、Bax、c leaved-Caspase-3和c leaved-Caspase-9的表达影响。构建Bc l-2的表达质粒,并用L ipo2000转染GBC细胞,并检测As2O3诱导细胞凋亡情况。结果:As2O3可诱导GBC细胞凋亡。蛋白水平的检测表明As2O3可使细胞Bc l-2、Bc l-xL的表达下降,B id、Bak和Bax表达升高,并促进Caspase-3和Caspase-9的剪切。过表达Bc l-2可抑制As2O3诱导的凋亡。结论:As2O3可诱导胆囊癌细胞的凋亡,并主要通过下调Bc l-2基因的表达来实现。
BACKGROUND & OBJECTIVE: Gallbladder cancer is a new and effective drug for the treatment of gallbladder cancer due to its lack of specific clinical manifestations, late diagnosis and low surgical resection rate; and the curative effect of conventional chemotherapy drugs on gallbladder cancer is poor. Arsenic trioxide has achieved breakthrough results in the treatment of acute promyelocytic leukemia. In recent years, it has attracted much attention in the study of solid tumors. This study aimed to investigate the biological effects of As2O3 on human gallbladder carcinoma GBC cell line and its mechanism. Methods: Hoechest 33258 staining, flow cytometry and DNA gel electrophoresis were used to study the apoptosis induced by As2O3. The effect of As2O3 on the expression of apoptosis-related proteins Bc 1-2, Bcl-xL, B id, Bak, Bax, c leaved-Caspase-3 and c leaved-Caspase-9 was analyzed using Western blot. The Bcl-2 expression plasmid was constructed and transfected into GBC cells with L-ipo2000, and the apoptosis induced by As2O3 was detected. Results: As2O3 induced GBC apoptosis. The protein level of As2O3 decreased the expression of Bcl-2 and Bcl-xL, increased the expression of B id, Bak and Bax, and promoted the cleavage of Caspase-3 and Caspase-9. Overexpression of Bcl-2 inhibited As2O3-induced apoptosis. Conclusion: As2O3 can induce the apoptosis of gallbladder carcinoma cells and mainly through down-regulating the expression of Bcl-2 gene.