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目的:探讨抗坏血酸(AA)对三氧化二砷(As2O3)诱导人多发性骨髓瘤(MM)细胞株RPMI8226细胞凋亡的作用及其分子机制。方法:采用细胞计数、细胞形态学、透射电镜、吖啶橙溴化乙锭(AOEB)染色、AnnexinⅤ染色以及细胞周期测定,观察As2O3联合AA对RPMI8226细胞的增殖抑制及促凋亡作用;应用流式细胞仪检测RPMI8226细胞死亡受体DR4分子的表达。结果:单用AA对RPMI8226细胞的生长无影响;As2O3抑制RPMI8226细胞的生长,其作用呈时间和剂量依赖性;As2O3联合AA较单用As2O3对RPMI8226细胞的抑制作用强(P<0.01)。单用5μmol/L的As2O3对靶细胞作用后,无明显凋亡形态学改变;100μmol/L的AA联合5μmol/L的As2O3作用靶细胞后:光镜、电镜和AOEB染色显示出典型的凋亡形态学改变;AnnexinV染色检测到早期凋亡细胞;细胞周期显示:G1期细胞比例增高,S期减低,并有凋亡峰。As2O3能上调RPMI8226细胞表达DR4分子,AA联合As2O3可以增强上述效应(P<0.05)。结论:AA对As2O3诱导RPMI8226细胞凋亡有增敏效应,该效应过程可能依赖于死亡受体DR4的过度表达。
Objective: To investigate the effect of ascorbic acid (AA) on the apoptosis of human multiple myeloma (MM) cell line RPMI8226 induced by As2O3 and its molecular mechanism. Methods: The proliferation and apoptosis of RPMI8226 cells treated with As2O3 and AA were observed by cell counting, cell morphology, transmission electron microscopy, AOEB staining, Annexin Ⅴ staining and cell cycle assay. Detection of DR4 molecule expression in RPMI8226 cell death receptor by cytometry. RESULTS: As2O3 alone had no effect on the growth of RPMI8226 cells. As2O3 inhibited the growth of RPMI8226 cells in a dose- and time-dependent manner. As2O3 combined with AA had stronger inhibitory effects on As2O3 than RPMI8226 cells alone (P <0.01). After treated with 5μmol / L As2O3 alone, there was no obvious apoptosis morphological changes. After treated with 100μmol / L AA combined with 5μmol / L As2O3, the apoptosis of the target cells was observed by light microscopy, electron microscopy and AOEB staining. Morphological changes; AnnexinV staining detected early apoptotic cells; cell cycle showed: G1 phase cells increased, S phase decreased, and the peak of apoptosis. As2O3 can up-regulate the expression of DR4 in RPMI8226 cells. AA combined with As2O3 can enhance the above effects (P <0.05). CONCLUSION: AA sensitizes the apoptosis of RPMI8226 cells induced by As2O3, which may depend on the overexpression of death receptor DR4.