维生素K3和三氧化二砷诱导多发性骨髓瘤细胞凋亡的协同作用

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。

Cooperation of Vitamin K3 and Apoptosis of Multiple Myeloma Induced by As2O3

Objective:In order to find the more effective way to treat MM,we explored the proliferation of inhibition and apotosis-inducing effect of VK3 and As2O3 alone or in combination on RPMI8226 cells.Methods:Growth inhibition of RPMI8226 cells exposed to Vitamin K3 combination with As2O3 was analyzed by MTT assay,apoptosis was evaluated by means of DNA gel electrophoresis,flow cytometry and Annexin V labeling.The expression of VEGF was analyzed with RT-PCR.Result:VK3 or As2O3 obviously inhibited the proliferation and induced apoptotic morphology and DNAfragmentation of MM cells.They also can reduce the level of bcl-2mRNA.Both of them showed synergic phenomenon.These effects were all time-dependent and dose-dependent.There was the character of apoptotic cells in RPMI8226 cells exposed to Vitamin K3 or As2O3 such as cellular atrophy,chromatin clustering,pyknosis and karyoklasis.Agarose gel electrophoresis of DNA samples from RPMI8226 cells exposed to Vitamin K3 or As2O3 showed obvious ladder strips.The expressions of VEGF and Bcl-2albumen at RPMI8226 cells treated with Vitamin K3 and As2O3 alone or in combination was decreased.Conclusion:VK3 and As2O3 alone or in combination obviously inhibited the proliferation of RPMI8226 cells,and showed apoptosis-inducing effect in a dose-dependent and a time-dependent manner.The expression of VEGF albumen at RPMI8226 cells treated with Vitamin K3 and As2O3 alone or in combination was decreased.Vitman K3 and As2O3 have synergy function to RPMI8226 cells.