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目的 探讨三氧化二砷 (As2 O3)对食管癌 Ec10 9细胞系的生物学效应及细胞和分子机制。方法 通过四氮唑 (MTT)还原法检测三氧化二砷 (As2 O3)对食管癌 Ec10 9细胞系存活率的影响 ,从形态观察、流式细胞仪分析、DNA凝胶电泳、细胞凋亡原位检测 (TU NEL)研究三氧化二砷诱导食管癌 Ec10 9细胞凋亡的情况 ,并以 Western blot检测基因表达。结果 Ec10 9细胞经 As2 O3处理后 ,存活率明显降低。光学显微镜下可见到明显的凋亡细胞 ,在细胞周期 G1期前有低于 2倍体的凋亡峰 ;DNA凝胶电泳显示出典型的凋亡特征 :DNA有规律断裂形成的梯状图谱 ,细胞凋亡原位检测发现 DNA断裂 ,Western blot检测表明 c- myc基因的表达下降。结论 As2 O3能诱导人食管癌Ec10 9细胞凋亡 ,并伴随 c- m yc基因的降调节。
Objective To investigate the biological effects and cellular and molecular mechanisms of arsenic trioxide (As2O3) on esophageal cancer Ec10 9 cell line. Methods The effect of arsenic trioxide (As2O3) on the survival rate of esophageal cancer Ec10 9 cell line was detected by tetrazolium (MTT) reduction method, from morphological observation, flow cytometry analysis, DNA gel electrophoresis, and apoptosis in situ detection ( TU NEL) was used to study the apoptosis of Ec10 9 cells induced by arsenic trioxide, and Western blot was used to detect gene expression. Results The survival rate of Ec10 9 cells after treatment with As2O3 was significantly reduced. Obvious apoptotic cells were observed under the light microscope. Apoptotic peaks below the diploid were observed before the G1 phase of the cell cycle; DNA gel electrophoresis showed a typical apoptotic characteristic: a ladder pattern of regular DNA breaks. The DNA fragmentation was detected by in situ apoptosis assay. Western blot analysis showed that the expression of c-myc gene was decreased. Conclusion As2O3 can induce apoptosis in human esophageal cancer Ec10 9 cells, accompanied by down-regulation of c-myc gene.