曲古霉素A对多发性骨髓瘤U266细胞株的影响  

Effects of trichostain A on multiple myeloma cell line U266

作　　者：许闪闪[1] 何丽梅[1] 李智[1] 翁文浩[1] 于慧[1] 

机构地区：[1]同济大学附属第十人民医院检验科,上海200072

出　　处：《白血病．淋巴瘤》2010年第7期394-397,共4页Journal of Leukemia & Lymphoma

摘　　要：目的 观察组蛋白去乙酰化酶抑制剂曲古霉素A（TSA）对多发性骨髓瘤U266细胞株增殖、周期、凋亡和侵袭力的影响.探讨该药对DNA甲基转移酶（DNMT）、甲基结合结构域（MBD）蛋白MBD2和MeCP2的影响.方法 不同浓度ISA作用于U266细胞株12、24、48和60 h,MTT法检测细胞的增殖活性,并计算药物作用24 h相应IC50.以IC50 作用U266细胞24 h后,检测细胞周期;荧光定量PCR分析基质金属蛋白酶-2（MMP-2）基因、抗凋亡基因bc1-2、bcl-xl以及DNMT、MBD2、MeCP2 mRNA的改变表达.分别用流式细胞术和Western blotting检测MMP-2和MBD2蛋白的变化.结果 TSA对U266细胞增殖的影响具有时间和剂量依赖性.作用24h的JC50为0.07μmol/L.TSA可将细胞阻滞于G0/G1期,细胞增殖指数（49.90±0.39）%,与对照组的（55.78±0.49）%相比差异有统计学意义（P〈0.01）.TSA作用后,MMP-2、bcl-2、bcl-xl 2-△△Ct 分别为：0.71±0.06、5.04±0.92、2.95±0.35,与对照组相比P〈0.05,TSA作用可以逆转DNMT、MBD2和MeCP2转录.结论 TSA可通过干扰细胞周期,将U266细胞阻滞于G0/G1期,从而抑制细胞增殖,促进凋亡的发生,该作用可能是通过改变细胞甲基化调节蛋白的表达而实现.Objective To evaluate effects of trichostain A （TSA） on cell proliferation, cell cycles, apoptosis and invasiveness of multiple myeloma cell line U266; as well as active changes of methylation regulating proteins including DNA methyl-transferase（DNMTs）, methyl-binding domain （MBD） proteins： MBD2 and MeCP2 after treated with TSA. Methods U266 cells were treated with different concentrations of TSA for 12, 24, 48 and 60 h. The proliferation activity of U266 cells was detected by MTT and the IC50 of 24 h was calculated. After U266 cells were treated with IC50, cell cycles were check out by dying with PI. mRNA of matrix metalloproteinase-2（MMP-2）, bc1-2, bcl-xl and methylation regulating proteins （DNMTs, MBD2 and MeCP2） were detected by real-time PCR. FCM and Western blotting were used to measure expressions of MMP-2 and MBD2. Results MTT results revealed TSA inhibited proliferation of U266 cells in a dose-and time-dependent manner and the IC50 of 24 h was 0.07 μmol/L FCM analysis showed that TSA could arrest the cell cycle in G0/G1 and the proliferation index （PI） in U266 cells [（49.90 0.39）%]were significantly different after exposed to TSA （0.7 μmnol/L for 24h compared with that in the control cells[（55.78 0.49）%]（P 〈0.01）. After treated by TSA, the 2-△△Ct of MMP-2, bcl-2 and bcl-xl were 0.71 0.06, 5.04 0.92 and 2.95 0.35, respectively. There were great changes on mRNA of DNMT, MBD2 and MeCP2. TSA could reverse the transcription of DNMT, MBD2 and MeCP2. Conclusion TSA can arrest the U266 cell cycle in GVG, to prevent its proliferation and promote apoptosis, which maybe greatly connect with the changes of the methylation regulating proteins.

关 键 词：多发性骨髓瘤 DNA甲基转移酶 细胞周期 曲古霉素A 

分 类 号：R733.3[医药卫生—肿瘤]