美伐他汀对人多发性骨髓瘤细胞系U2 66体外增殖与凋亡的影响  

In vitro Effects of Mevastatin on the Proliferation and Apoptosis in Human Multiple Myeloma Cell Line U266

作　　者：刘泽林[1] 罗建民[1] 董作仁[1] 王福旭[1] 张学军[1] 杨敬慈[1] 杜行严[1] 姚丽[1] 

机构地区：[1]河北医科大学第二医院血液科

出　　处：《中国实验血液学杂志》2004年第3期340-345,共6页Journal of Experimental Hematology

摘　　要：为了探讨羟甲基戊二酸单酰辅酶A(HMG CoA)还原酶抑制剂对多发性骨髓瘤 (MM)细胞增殖与凋亡的调控作用及机制 ,应用MTT比色法、光镜形态学观察、流式细胞术、DNA凝胶电泳及RT PCR研究了美伐他汀(mevastatin )对MM细胞系U2 6 6细胞体外增殖与凋亡的影响。结果显示 ,美伐他汀以剂量及时间依赖方式抑制U2 6 6细胞增殖 ;细胞周期分析显示美伐他汀诱导U2 6 6细胞G0 -G1 期阻滞 ,但对 p2 7蛋白及mRNA的表达无影响 ;在美伐他汀作用下U2 6 6细胞出现核染色质凝聚、核固缩、核碎裂等凋亡形态改变 ,PI- AnnexinⅤ + 细胞增多 ,线粒体跨膜电位 (△ψm)下降 ,DNA凝胶电泳出现梯形条带 ,bcl 2mRNA表达下调。加入外源性甲羟戊酸 (meva lonate,MVA)可完全逆转美伐他汀对U2 6 6细胞增殖与凋亡的效应。结论 :美伐他汀可通过MVA途径 ,诱导G0-G1 期阻滞抑制增殖、下调bcl 2表达及降低线粒体膜电位触发U2 6 6细胞凋亡。In order to investigate the anti-tumor activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors and the mechanism underlying the cell proliferation and apoptosis modulated in myeloma cells,the effects of mevastatin,an HMG-CoA reductase inhibitor,on cell growth,cell cycle progression and apoptosis in U266 human multiple myeloma (MM) cell line in vitro were explored by MTT colorimetric assay,morphologic observation,flow cytometry,DNA gel electrophoresis,and RT-PCR. The results demonstrated that mevastatin inhibited the growth of U266 cells in time- and dose-dependent manners. Cell cycle analysis showed that U266 cells underwent G 0/G 1 arrest under exposure to mevastatin,but it did not affect p27 expression at both mRNA and protein level. Morphologic observations revealed cytoplasm shrinkage,nuclear condensation and fragmentation in mevastatin-treated cells,and fraction of annexinⅤ+PI- cells increased significantly in the presence of the agent as determined by flow cytometric assay. In addition,mevastatin caused the collapse of mitochondrial transmembrane potential (Δψm),induced DNA fragmentation,and down-regulated the mRNA expression of bcl-2. The growth-inhibitory,cell cycle arresting,and proapoptotic effects of mevastatin in U266 cells could be effectively reversed by the addition of mevalonate (MVA),the immediate endproduct of the reaction catalyzed by HMG-CoA reductase. It is concluded that mevastatin suppresses proliferation by inducing G 0/G 1 phase arrest and triggering apoptosis via down-regulation of bcl-2 and reduction of Δψm,which may be attributed to the inhibition of MVA pathway by mevastatin. Statins including mevastatin may find their future application in the treatment of MM.

关 键 词：羟甲基戊二酸单酰辅酶A还原酶 美伐他汀 多发性骨髓瘤 U266细胞 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]