PMA诱导K562细胞向单核细胞分化机制探讨  

Signaling mechanism of PMA-induced monocytic differentiation of K562

作　　者：潘素飞[1,2] 任霞[2] 史美燕 禹林昌 张之勇[2] 姜国胜[2] 

机构地区：[1]济南大学.山东省医学科学院医学与生命科学学院,山东济南250062 [2]山东省医学科学院基础医学研究所.山东省现代医用药物与技术重点实验室.山东省医药卫生肿瘤免疫与中药免疫重点实验室,山东济南250062

出　　处：《中华肿瘤防治杂志》2014年第14期1049-1053,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81172792);山东省自然科学基金(ZR2011HL045;ZR2011HL050);山东省卫生厅青年基金(2007QZ023);山东省中医药科技发展计划(2011-234)

摘　　要：目的:探讨佛波酯(phorbol-12-myristate-13-ace-tate,PMA)诱导K562细胞向单核/巨噬细胞分化的作用机制。方法:采用CCK8法检测0、6.25、12.5、25、50、100和200nmol/L PMA对K562细胞增殖的影响,应用Wright-Gimesa染色观察细胞形态学变化,采用流式细胞术检测细胞表面分化抗原CD11b和CD14的表达变化,采用RT-PCR分析TGF-β1及其下游基因SMAD3和SMAD4在基因水平的表达趋势,并采用蛋白质印迹法检测TGF-β1在蛋白水平的表达变化。结果:K562细胞经6.25nmol/L PMA处理后72h增殖抑制率为(22.03±2.7)%,12.5nmol/L为(31.04±4.3)%,25nmol/L为(35.03±3.5)%,50nmol/L为(47.01±4.1)%,100nmol/L为(55.06±5.2)%,200nmol/L为(76.72±5.4)%,差异有统计学意义,F=2.24,P<0.05。PMA能抑制K562细胞的增殖并能促进其分化,作用效果随剂量的加大逐渐增强;细胞形态学上趋向于成熟分化;细胞表面分子CD11b和CD14的表达量升高。在mRNA水平,TGF-β/SMAD信号通路因子TGF-β1的表达量随时间的延长逐渐升高其下游因子SMAD3和SMAD4的表达也呈上升趋势;在蛋白水平,TGF-β1的表达亦呈上升趋势。结论:PMA可通过促进TGF-β/SMAD信号的传导诱导白血病细胞株K562细胞向单核/巨噬细胞分化。OBJECTIVE： To explore the mechanism of PMA induce K562 cells differentiated to monocyte-macropha- ges. METHODS：The cell counting Kit 8（CCK8） assay was used to detect the effects of PMA on K562 cells proliferation. The cell morphology changes were observed by Wright-Giemsa staining. The cell surface markers CD11b,CD14 were ana- lyzed by flow cytometry. The expression of TGF-β1 ,SMAD3 ,and SMAD4 were detected by RT-PCR and the protein level of TGF-β1 was detected by western blot. RESULTS： PMA can inhibit the proliferation of K562 cells,and can promote the differentiation by dose dependent. The inhibition rates of 6.25,12.5,25,50,100 and 200 nmol/L PMA were （22.03± 2.7）%,（31.04±4.3）% ,（35. 03±3. 5）%,（47.01±4. 1）%,（55.06±5. 2）% and （76.72±5.4）% respectively（F= 2.24, P〈0.05）. Wright-Giemsa staining indicated that the numbers of mature monocytes were increased. The expression of the cell surface markers CD11b,CD14 was increased. In mRNA levels, the expression of TGF-β/SMAD signaling path- way factor TGF-β1 and its＇ downstream factors SMAD3,SMAD4 were obviously up-regulated. The expression of TGF-β1 was also up-regulated in protein levels. CONCLUSION： PMA may promote the differentiation of K562 cells by influencing the transduction of TGF-β/SMAD signaling pathway.

关 键 词：佛波酯 K562 分化 TGF-Β SMAD 细胞增殖 

分 类 号：R733.7[医药卫生—肿瘤]