自噬对多发性骨髓瘤细胞增殖的影响  

Role of autophagy on proliferation of multiple myeloma cells and its possible mechanism

作　　者：高灵素[1] 虞国慧[1] 

机构地区：[1]安徽省六安市人民医院,安徽六安215004

出　　处：《安徽医药》2015年第4期636-640,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨自噬(autophagy)对正常培养多发性骨髓瘤(Multiple myeloma,MM)细胞株增殖的影响。方法 MM细胞株U266在正常培养条件下分别加入雷帕霉素和3-甲基腺嘌呤(3-methyladenine,3-MA),以正常血清培养U266细胞及淋巴瘤Jurkat细胞株为对照组;分别检测细胞的增殖及凋亡水平;单丹磺酰尸胺(monodansylcadaverine,MDC)荧光染色法检测细胞自噬水平;逆转录PCR检测自噬相关基因mtor、Beclin1、Atg5在细胞内水平,蛋白质印迹法检测细胞内自噬相关蛋白轻链3I/Ⅱ(LC3I/Ⅱ)含量。结果 U266细胞及Jurkat细胞内均存在自噬,但前者水平高于后者。雷帕霉素可诱导MM细胞发生自噬,3-MA对自噬有双重作用。雷帕霉素及3-MA下调细胞增殖水平。雷帕霉素及3-MA处理24 h可上调正常培养条件下U266细胞凋亡水平(1.7%±0.2%,19.1%±1.0%和16.8%±0.61%)。延长培养时间至72 h,雷帕霉素处理组凋亡水平无明显变化(16.9%±0.48%)。而3-MA处理组凋亡水平下降(9.0%±0.70%)。结论 MM细胞株内会存在一定水平的自噬,并通过实验证明其高于淋巴瘤Jurkat细胞株的自噬水平。在正常培养条件下加入了雷帕霉素及3-MA可抑制MM细胞的增殖,并可诱导细胞凋亡。Objective To investigate the influence of autophagy on the proliferation of multiple myeloma （ MM） cells under normal cul-ture condition.Methods Multiple myeloma （ MM） cell line U266 was incubated with rapamycin or 3-MA respectively.Normally cul-tured Jurkat and U266 cell were used as control group.We determined the proliferation and apoptosis of cells respectively.Monodansyl-cadaverine（ MDC） staining was employed to detect the autophagy.Reverse transcription PCR detected cells autophagy related genes mTOR,Beclin1,Atg5 level.Western blot was employed to detect the expression of protein LC3I /LCII.Results Autophagy level in U266 cells was higher than Jurkat cells.Rapamycin induced autophagy in U266 cell .3-MA showed inhibitory effect on autophagy for a short time.Rapamycin or 3-MA could induce cell apoptosis when incubated in normal culture condition for 24 hour （19.1%±1.0 %and 16.8%±0.61%,control group 1.7%±0.2%）.Prolonged incubation time to 72 hours,the apoptosis level of rapamycintreatment group had no difference（16.9%±0.48%）.But the levels of apoptosis of 3-MA group was decreased（9.0%±0.70%）.Conclusion There was a basic level of autophagy in MM cell,and it was higher than those in the Jurkat cells.Rapamycin and 3-MA both inhibited the proliferation of MM cell and induced cell apoptosis.

关 键 词：多发性骨髓瘤 雷帕霉素 3-甲基腺嘌呤 自噬 细胞凋亡 

分 类 号：R733.3[医药卫生—肿瘤]