5-氮杂-2'-脱氧胞苷联合古曲霉素A对人多发性骨髓瘤细胞系RPMI8226细胞增殖、凋亡及DLC-1基因表达的影响(英文)  

Effect of 5-Aza-2'-Deoxycytidine Combined with Trichostatin A on RPMI-8226 Cell Proliferation,Apoptosis and DLC-1 Gene Expression

作　　者：郭靖[1] 冯献启[1] 聂淑敏[2] 苏湛[1] 史雪[1] 崔忠光 张玲[1] 刘世国[3] 孟凡军[1] 赵春亭[1] 

机构地区：[1]青岛大学医学院附属医院血液内科,青岛256600 [2]青岛大学医学院附属医院神经科,青岛256600 [3]青岛大学医学院附属医院痛风实验室,青岛256600

出　　处：《中国实验血液学杂志》2014年第2期357-363,共7页Journal of Experimental Hematology

基　　金：山东省自然科学基金(ZR2010HM094)

摘　　要：本研究旨在探讨DNA甲基化抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-cdR)联合组蛋白去乙酰化抑制剂古曲霉素A(TSA)对人多发性骨髓瘤细胞株RPMI 8226细胞的生物学活性及DLC-1基因表达调控的影响。5-Aza-CdR及TSA单独或联合作用于RPMI 8226细胞系,用CCK-8法检测细胞增殖活性,实时定量PCR(RT-PCR)检测药物作用后DLC-1基因表达情况,ELISA检测药物作用后RhoA及Rac1蛋白表达水平。结果表明,5-Aza-CdR及TSA对多发性骨髓瘤细胞具有明显生长抑制作用并呈剂量依赖性(P<O.05);与5-Aza-CdR及TSA单药组比较,联合用药组对细胞的抑制作用更强,细胞凋亡率明显升高(P<0.05);对照组RPMI 8226细胞DLC-1基因微弱表达,5-AzacdR组DLC-1基因表达增强,且呈剂量依赖性重新表达(P<0.05),TSA虽不能诱导基因重新表达,但两药联合应用时可明显增强细胞DLC-1基因的表达;实验组与对照组多发性骨髓瘤细胞相比,rhoA及Rac1蛋白表达明显下降(P<0.05)。结论:5-Aza-CdR及TSA能有效的逆转RPMI 8226细胞DLC-1基因的表达,并明显增强对多发性骨髓瘤细胞的生长抑制及促凋亡作用。这种抑制作用可能与其抑制Rho/ROCK信号途径有关。This study was aimed to investigate the effects of the DNA methylation inhibitor 5-aza-2＇-deoxycytidine （5-Aza-CdR） and histone deacetylase inhibitor trichostatin A （TSA） on DLC-1 gene transcription regulation and molec- ular biological behaviours in the human multiple myeloma RPMI-8226 cells. The cells were treated respectively with 5- Aza-CdR and TSA alone, or the both combination; the cell proliferation and apoptosis, DLC-1 expression, the protein expression of Ras homolog family member A （RhoA） and Ras-related C3 botulinum toxin substrate 1 （Racl） were ex- amined by CCK-8 method, RT-PCR and ELISA, respectively. The results showed that the 5-Aza-CdR and TSA had cell growth inhibitory and apoptosis-inducing effects in dose-dependent manner （ P 〈 0.05 ）. Compared with a single drug （5-Aza-CdR or TSA alone）, the effects were significantly enhanced after treatment with the combination of 5-Aza-CdR and TSA （ P 〈 0.05 ）. DLC-1 was weakly expressed in the control group; the treatment with 5-Aza-CdR alone enhanced its re-expression dose-dependently （ P 〈 0.05 ）. Compared with 5-Aza-CdR alone, 5-Aza-CdR plus TSA enhanced DLC- 1 re-expression significantly. Compared with the control, 5-Aza-CdR and TSA significantly decreased RhoA and Racl protein expression （P 〈 0.05 ）. It is concluded that 5-Aza-CdR and TSA can effectively reverse DLC-1 expression of RPMI-8226 cells; TSA has a synergistic effect on its re-expression. 5-Aza-CdR and TSA have significant cell growth in- hibitory and apoptosis-inducing effects on RPMI-8226 cells. These effects may be related to the inhibition of Rho/Rho kinase signalling pathway.

关 键 词：多发性骨髓瘤 5-氮杂-2’-脱氧胞苷 曲古抑菌素A RPMI-8226细胞 DLC-1基因 

分 类 号：R733.3[医药卫生—肿瘤]