二甲双胍通过调节成脂影响脂肪细胞对白血病细胞作用  

Metformin Influences the Effect of Adipocytes on Leukemia Cell Chemoresistance through Regulation of Adipogenesis

作　　者：王慧[1] 刘四红[2] 翟元梅[1] 朱晓宇[1] 章菁[2] 万云[1] 路伟[1] 石军[1] 

机构地区：[1]上海交通大学附属第六人民医院血液科,上海200233 [2]苏州大学医学部,江苏苏州215123

出　　处：《中国实验血液学杂志》2015年第2期340-344,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(81170507);科技部重大仪器专项子课题(2013YQ03065109);上海市科委项目(14142200702);上海市科委课题(13DZ2293702)

摘　　要：目的:探讨二甲双胍对3T3-L1前脂肪细胞的诱导分化作用,并研究处理后脂肪细胞对白血病细胞的影响。方法:在3T3-L1前脂肪细胞诱导分化成熟过程中分别给予不同浓度二甲双胍处理,诱导后通过油红O染色和吸光度值(OD值)检测其分化程度;应用real-time PCR检测成脂后关键基因PPARγ、C/EBPα、FABP4(ap2)mRNA的表达水平。诱导后的脂肪细胞分别与GFP+-THP-1细胞共培养,加入1μg/ml的阿糖胞苷,48 h后采用流式细胞仪检测GFP+-THP-1细胞的凋亡。收集诱导后脂肪细胞的上清,与肿瘤细胞基础培养液(RPMI1640)以1∶1混合后培养肿瘤细胞,应用CCK8检测脂肪细胞对白血病细胞增殖,加入1μg/ml的阿糖胞苷,48 h后应用细胞计数仪检测其化疗保护作用。结果:二甲双胍处理组OD值及成脂基因C/EBPα、FABP4表达较对照组低,而PPARγmRNA表达无明显改变;二甲双胍处理后的脂肪细胞共培养组白血病细胞凋亡率高于对照组。脂肪细胞促进白血病细胞增殖并对其有化疗保护作用,这些作用被二甲双胍降低。结论:二甲双胍能抑制3T3-L1前脂肪细胞的成脂分化,并调节脂肪细胞对白血病细胞凋亡、增殖及化疗的保护作用。Objective： To investigate the effect of metformin on 3T3-L1 preadipocyte＇ s differentiation and consequently observe the anti-proliferative effects of metformin-treated adipocytes on leukemia cells. Methods： Different concentrations of mefformin were added in 3T3-L1 preadipocytes to induce maturation, the matured adipocytes were detected by oil red O staining and quantified by absorbance value （OD）. Real-time PCR was used to detect the mRNA expression level of the key adipogenic genes PPAR）,, C/EBPa and FABP4 （ ap2 ）. The adipocytes were co-cultured with GFP -THP-1 cells, 1 μg / ml of cytarabine（Ara-C） was added and incubated for 48 hours, the flow cytometry was used to detect the apoptosis rate of GFP - THP-1 cells. Adipocyte supernatant was collected and mixed with equal volume of tumor lysat medium （RPMI 1640） at 1：1 to culture tumor cells. The leukemia cell proliferation activity was detected by CCK-8; after 48 hours of adding 1 μg/ml Ara-C, the protective effect on chemotherapy was assayed by using cytometer. Results： The metformin lowered the OD value, and the expression levels of both adipogenic genes C EBPa and FABP4 were lower than those of controls, while the expression level of PPARy mRNA was not significantly changed, the apoptosis rate of leukemia cells co-caltured with metformin-treated adipocytes was higher than that of cocultured cells without metformin treatment. The adipocytes promoted the leukimia cell proliferation and protected leukemia cells from chemotherapy, which could be abrogated by mefformin. Conclusion： The mefformin can inhibit the differentiation of 3T3-L1 preadipocytes into adipocytes, and can regulate the protective effect of adipocytes on the apoptosis, proliferation and chemotherapy of leukemia cells.

关 键 词：二甲双胍 3T3-L1前脂肪细胞 脂肪细胞 白血病细胞 细胞增殖 细胞凋亡 化疗保护 

分 类 号：R733.7[医药卫生—肿瘤]