摘要
目的 探讨丹参酮ⅡA(tanshinoneⅡA, TanⅡA)对耐维甲酸的急性早幼粒细胞白血病(APL)细胞诱导分化作用及其分子机制。方法 以0 5mg/LTanⅡA处理APL细胞株NB4细胞作为阳性对照,将耐维甲酸的APL细胞株MR 2细胞与1 0mg/LTanⅡA在体外共同培养4d,观察细胞生长状况、形态变化,并检测药物作用前后细胞四氮唑蓝(NBT)还原能力,用流式细胞术(FCM)测定细胞增殖周期、c myc、bcl 2、p53、c fos、CD33及CD11b表达。结果 1 0mg/LTanⅡA能明显抑制MR 2细胞生长(P<0 01),生长抑制率为73 5%, 0 5mg/LTanⅡA能抑制NB4细胞生长(P<0 01),抑制率为67 7%,TanⅡA对两者的抑制作用差异无统计学意义(P>0 05)。经TanⅡA处理后,MR 2、NB4细胞形态趋于成熟粒细胞,表现为细胞体积变小,核浆比例缩小,染色质变粗糙,核仁消失,细胞质内嗜苯胺蓝颗粒消失,核形态不规则,可见晚幼粒细胞,且TanⅡA处理NB4细胞后可见杆状核粒细胞。NBT还原实验显示,TanⅡA处理MR 2、NB4组阳性率分别为( 95 30±0 76 )%、(93 20±1 04)%,而相应空白对照组分别为(3 50±1 32)%、( 2 80±0 29 )%,处理组高于对照组(P<0 01)。FCM分析发现,TanⅡA处理MR 2、NB4细胞后CD33表达下降,CD11b表达升高;处理后的细胞G0 /G1 期比例增高,S期细胞降低。
Objective To investigate retinoic acid -resistant acute promyelocytic leukemia (APL) cell differentiation induced by tanshinone ⅡA (Tan ⅡA ) and its molecular mechanism Methods NB4 cells treated with 0 5 mg/L TanⅡA was regarded as positive control After in vitro incubation of MR-2 cells with Tan ⅡA at the concentration of 1 0 mg/L for 4 days ,the cell differentiation was observed by growth status, cytomorphology, and nitroblue tetrazolium test Cell cycle, membrane cluster differentiation (CD) antigens (CD_ 33 , CD_ 11b ) and expression of some oncogene (c-myc, c-fos, p53 and bcl-2) were analysed by flow cytometry Results The growth of MR-2 and NB4 cells was inhibited after TanⅡA treatment, the inhibition rate were 73 5% and 67 7% respectively ( P <0 01, P <0 01) without significant difference After TanⅡA treatment, MR-2 and NB4 cells were induced to undergo morphological differentiation, which exhibited small cell bulk decreased nucleus/cytoplasm proportion, rough chromatin, disappearence of nucleolus and formation of azurophil granules and anomalous nucleus MR-2 cells could be induced to metamyelocyte while NB4 could be induced to band form NBT reduction of MR-2 and NB4 cells treated with TanⅡA showed that positive cells accounted for (95 30±0 76)% and(93 20±1 04)% respectively; but the positive rate of either group of the treated positive cells was significantly higher than that of untreated, being (3 50±1 32)% and (2 80±0 29)% respectively ( P <0 01) Flow cytometry showed that the expression of CD_ 33 was reduced, while that of CD_ 11b was increased The quantity of treated cells in G_0/G_1 phase increased but that in S phase decreased The proliferous index was also decreased After treated with TanⅡA, the expressions of anti-oncogene p53 and c-fos were up-regulated while those of oncogene bcl-2 and c-myc were down-regulated ( P <0 01) Conclusions 1 0 mg/L TanⅡA could inhibit proliferation of MR-2 cells and ind
出处
《中华内科杂志》
CAS
CSCD
北大核心
2005年第5期366-369,共4页
Chinese Journal of Internal Medicine
基金
天津市自然科学基金资助项目(013615611)
