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目的:探讨三氧化二砷(As2O3)联合维生素K3对人多发性骨髓瘤细胞株RPMI8226细胞生长抑制及诱导凋亡作用机制。方法:采用四甲基噻唑氮蓝比色法检测As2O3联合VK3对RPMI8226细胞生长抑制作用、DNA电泳、流式细胞术Annexin V/PI标记法检测凋亡,观察As2O3联维生素K3对RPMI8226细胞的增殖抑制及促凋亡作用;同时通过RT-PCR检测VEGF表达来观察VK3、As2O3对RPMI8226细胞的凋亡诱导作用及两者是否有协同作用。结果:VK3和As2O3二者均可明显抑制RPMI8226细胞系的增殖,其作用机制可能通过降低Bcl-2mRNA的表达、诱导调亡,这种效应具有时间与剂量依赖性;二者间具有协同作用。经VK3和As2O3作用后的RPMI8226细胞出现凋亡细胞的特征;维生素K3(VK3)、As2O3单独及联用作用于RPMI8226细胞DNA凝胶电泳可见明显的梯形条带。与单用As2O3相比,As2O3联合VK3作用于RPMI8226细胞48h后,细胞凋亡率显著增加(P<0.05),G0/G1期细胞比例升高,S期细胞减少。RT-PCR检测表明RPMI8226细胞经VK3、As2O3单独及联合作用后血管内皮生长因子VEGF表达明显降低。结论:VK3、As2O3单用及联用对RPMI8226细胞有明显的增殖抑制作用和诱导凋亡作用,且具有一定的量效和时效关系。可能通过VEGF表达降低而起作用。VK3和As2O3在诱导RPMI8226细胞凋亡中具有明显的协同作用。
Objective: To investigate the mechanism of arsenic trioxide (As2O3) combined with vitamin K3 on the growth inhibition and induction of apoptosis in human multiple myeloma RPMI8226 cell line. Methods: The inhibitory effects of As2O3 and VK3 on the growth of RPMI8226 cells were detected by MTT assay. The apoptosis of RPMI8226 cells was detected by DNA electrophoresis and Annexin V / PI staining. The proliferation of RPMI8226 cells was observed by As2O3 combined with vitamin K3 Inhibition and pro-apoptotic effect. At the same time, VEGF expression was detected by RT-PCR to observe the apoptosis-inducing effect of VK3 and As2O3 on RPMI8226 cells and whether there is a synergistic effect between them. Results: Both VK3 and As2O3 could significantly inhibit the proliferation of RPMI8226 cell line. The mechanism may be through the decrease of Bcl-2 mRNA expression and induction of apoptosis. The effect is time-and dose-dependent; the synergistic effect exists between them. Apoptotic cells were found in RPMI8226 cells treated with VK3 and As2O3, and obvious trapezoidal bands were observed by DNA gel electrophoresis of vitamin K3 (VK3) and As2O3 in RPMI8226 cells alone and in combination. Compared with As2O3 alone, the apoptosis rate of RPMI8226 cells treated with As2O3 and VK3 for 48 h increased significantly (P <0.05), the proportion of cells in G0 / G1 phase increased and the number of S phase cells decreased. The results of RT-PCR showed that the expression of vascular endothelial growth factor VEGF in RPMI8226 cells treated with VK3 and As2O3 alone and in combination decreased significantly. CONCLUSION: VK3 and As2O3 can significantly inhibit the proliferation and induce apoptosis of RPMI8226 cells in vitro and in vitro, and have a certain dose-effect and time-effect relationship. It may play a role through the decrease of VEGF expression. VK3 and As2O3 have obvious synergistic effects in inducing RPMI8226 cell apoptosis.