丹参酮Ⅱ_A诱导白血病NB4细胞分化分子机制研究

Study of molecular mechanism of tanshinone Ⅱ_A inducing differentiation in acute promyelocytic leukemia NB4 cells

目的:探讨丹参酮ⅡA诱导白血病NB4细胞分化的分子机制。方法:0.5 mg.L-1丹参酮ⅡA体外处理NB4细胞72 h后,分别采用显微镜观察细胞形态分化;MTT检测细胞增殖抑制作用;收集细胞并提取总RNA,反转录获得cDNA,再转录并标记成cRNA(Cy3荧光标记),与Express ChipTMH04芯片杂交,最后对芯片进行扫描和数据分析,检测丹参酮ⅡA诱导NB4细胞分化前后相关基因谱表达的变化。结果:0.5 mg.L-1丹参酮ⅡA可诱导92.8%NB4细胞向终末细胞分化。其中,中、晚幼粒细胞占27.0%;杆状及分叶核粒细胞占68.2%;细胞生长被明显抑制。基因芯片检测发现:3 360条基因中有183条基因差异表达,其中包括23条(5条上调和18条上调)分化相关基因和与细胞凋亡、周期调控、DNA转录、DNA损伤/修复、蛋白转运、信号传导、核受体、细胞因子和生长因子、癌基因和抑癌基因等相关基因。结论:丹参酮ⅡA可能通过调控多种相关基因、特别是分化相关基因的表达诱导白血病细胞分化,本研究有助于阐明丹参酮ⅡA诱导分化抗肿瘤的分子机制。

Objective： To investigate molecular mechanism of tanshinone Ⅱ A inducing differentiation and apoptosis in acute promyelocytic leukemia NB4 cells. Method： NB4 cells were cultured in vitro and treated with tanshinone ⅡA and observed cellular morphology, cell category and the cellular proliferation. DNA microarray technique was used to analyze the gene expression profiles of NB4 ceils induced by tanshinone ⅡA. Result： 92. 8% of NB4 ceils treated with 0. 5 mg·L^-1 tanshinone ⅡA were induced into mature neutrophils, in which myetocytes and melamyetocytes were 27.0%, banded and segmented neutrophits 68.2%. Cell growth were inhibited. cDNA microarray showed the enormously expressed 183 genes including 23 differentiation associated genes, and other interrelated genes. Conclusion： Tanshinone ⅡA inducing differentiation in NB4 cells may be via regulation of many kinds of genes, especially differentiation associated genes expression. This partially explained the molecular mechanism of tanshinone ⅡA inducing differentiation.