摘要
目的 探讨丹参酮ⅡA(TanshinoneⅡA ,TanⅡA)对急性早幼粒细胞白血病 (APL)细胞诱导分化的作用及其分子机制。方法 将APL细胞株NB4细胞与TanⅡA在体外共同培养 5天 ,观察细胞形态变化 ,并检测药物作用前后细胞NBT还原能力 ,用流式细胞术测定细胞增殖周期、c myc、bcl 2、p5 3、c fos、CD3 3 及CD11b表达。结果 0 .5 μg/mlTanⅡA可诱导 (91.3± 2 .1) %的NB4细胞向终末细胞分化。其中 ,中、晚幼粒细胞占 0 .2 6 ,杆状及分叶核细胞占 0 .6 8;细胞生长明显抑制 ;NBT还原能力显著增强 ;CD3 3 表达下降 ,CD11b表达升高 ;与全反式维甲酸 (ATRA)相比 ,差异无显著性 (P >0 .0 5 )。流式细胞术分析发现 ,TanIIA处理组细胞被阻滞于G0 /G1期 ,S期细胞数明显减少 ,增殖指数降低 ,c myc、bcl 2基因蛋白表达降低 ,c fos、p5 3基因蛋白表达增加。结论 TanⅡA体外能使NB4细胞分化成熟 ,增殖能力下降。其机制可能是通过调控与NB4细胞增殖、分化相关的基因表达 ,抑制细胞DNA合成 ,从而抑制细胞增殖 ,诱导细胞分化。
Objective To investigate APL cell differentiation induced by tanshinone Ⅱ(Tan ⅡA) and its molecular mechanism. Methods In vitro incubation of NB4 cells with Tan ⅡA at the concentration of 0.5 μg/ml for 5 days,the cell differentiation was observed by cytomorphology,and nitroblue tetrazolium (NBT) test. Cell cycle, membrane CD 33 , CD 11b antigens and gene expressions (c myc, c fos, p53 and bcl 2) were analysed by flow cytometry. Results (91.3±2.1) % of NB4 cells were induced into morphologically and functionally more differentiated cells including 0.26 of myelocytes and metamyelocytes, and 0.68 of band form and neutrophils. Cell growth curve showed that growth of NB4 cells were inhibited. NBT reduction was significantly increased.Expression of CD 33 decreased and CD 11b increased.The degrees of cell differentiation and growth inhibition induced by Tan ⅡA or ATRA were no difference. Flow cytometry analysis showed that Tan ⅡA arrested NB4 cell in G 0/G 1 phase, inhibited cellular DNA synthesis, down regulated c myc and bcl 2 genes expression, and up regulated c fos and p53 genes expression. Conclusion Tan ⅡA can induce differentiation and growth inhibition of NB4 cells. Its possible molecular mechanism might relate to modulation of gene expressions associated proliferation and differentiation, and to inhibition of DNA synthesis.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2000年第1期23-26,共4页
Chinese Journal of Hematology
基金
卫生部科学研究基金!96 1 2 40
关键词
丹参酮
白血病
细胞分化
分子机制
Tanshinone
Leukemia,promyelocytic, acute
Cell line,NB4
