三氧化二砷对黑色素瘤B-16细胞增殖与凋亡的影响

Effects of Arsenic trioxide on cell proliferation and apoptosis of melanoma B-16 cells

目的：观察三氧化二砷（As2O3）对体外培养黑色素瘤B—16细胞增殖和凋亡的影响。方法：应用Mrr比色法检测细胞增殖活力；采用流式细胞术测定细胞周期及凋亡率；倒置显微镜、激光共聚焦显微镜和透射电子显微镜观察细胞形态学变化和凋亡特征。结果：6．25～100μmol／LAs2O3作用细胞24、48小时后，细胞增殖活力明显下降，且具有时一效和量一效依赖关系；50μmol／LAs2O3作用细胞12小时后。细胞凋亡率增加。S期细胞百分率下降；倒置显微镜下可见细胞生长密度减小、形态变圆、体积缩小；hoechst33258核染色可见核染色质边集、浓集；电子显微镜下可见到典型的凋亡细胞。结论：三氧化二砷能抑制黑色素瘤B16细胞的增殖并可诱导其发生凋亡.

Objective：To investigate the effect of Arsenic trioxide on the cell proliferation and apoptosis of cultured melanoma B - 16 cells. Methods： MTT reduction assay was used to reflect the proliferation of the cells. The apoptosis rate and the cell cycle change were detected by flow cytometry. The morphological changes and apoptotic characteristics of the cells were observed with the aid of inverted microscope, LSCM and TEM. Results：Arsenic trioxide （6.25 to 100μmol/L with 24h and 48h duration） notably inhibited the growth of B - 16 cells, which showed the dependence relationship between time effect and dose effect, Cultured with 50 μmol/L Arsenic trioxide for 12h, flow cytometry analysis showed higher cell apoptosis rate and lower S phase percentage than those of the control group. Less cell growth density, cell rounding and c：ell shrinkage could be seen under inverted microscope; Chromatin margination and condensation appeared via hoechest dye, The apoptosis body, cell mitochondrium vaculation and deformation could be seen under the TEM. Conclusion：Arsenic trioxide can remarkably inhibit proliferation and induce apoptosis of melanoma B - 16 cells.