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目的 研究三丁酸甘油酯 (TB)与全反式维甲酸 (ATRA)不同浓度组合对NB4及MR2细胞的诱导分化作用 ,并观察TB对NB4、MR2细胞是否具有促凋亡作用。方法 通过硝基四唑氮蓝(NBT)还原率及细胞表面抗原CD11b、CD14、CD33表达观察细胞分化程度 ;通过细胞形态、DNA片段化电泳、流式细胞术 (FCM)DNA含量分析及凋亡细胞原位标记 (TUNEL)试剂盒检测细胞凋亡 ;用逆转录 聚合酶链反应 (RT PCR)检测bcl 2表达水平。结果 0 .2mmol/LTB联合不同浓度的ATRA对NB4细胞具有协同诱导分化效应 ;ATRA单用及联合 0 .2mmol/LTB对MR2细胞诱导分化作用不明显。TB 1 .0mmol/L作用 2 4h ,NB4及MR2细胞均出现典型的细胞凋亡形态 ;DNA凝胶电泳出现片段化的梯形带谱 ;FCM检测显示TB同时可影响NB4及MR2细胞的细胞周期进程 ,诱导细胞凋亡 ,而且亚G1期及G1期细胞所占比例相似 ;TUNEL检测亦证实了这一结果。TB在诱导细胞凋亡过程中随作用时间的延长 ,bcl 2基因的表达水平逐渐下降。结论 TB具有协同维甲酸诱导早幼粒细胞白血病细胞分化及致细胞凋亡的双重作用。
Objective To investigate the effects of different concentrations of tributyrin (TB) and all-trans retinoic acid (ATRA) on the differentiation of NB4 and MR2 cells, and to observe whether TB has an apoptosis-promoting effect on NB4 and MR2 cells. Methods The degree of cell differentiation was observed by the reduction of nitrotetrazolium blue (NBT) and the expression of cell surface antigens CD11b, CD14, and CD33. The cell morphology, DNA fragmentation electrophoresis, flow cytometry (FCM) DNA content analysis and apoptosis were analyzed. Apoptosis was detected by TUNEL kit; bcl 2 expression was detected by reverse transcription polymerase chain reaction (RT PCR). RESULTS: O.2mmol/LTB combined with different concentrations of ATRA had a synergistic effect on differentiation of NB4 cells; ATRA alone or in combination with 0.2mmol/LTB had no obvious effect on the differentiation of MR2 cells. After treated with TB 1 .0mmol/L for 24 hours, NB4 and MR2 cells showed typical apoptotic morphology; DNA fragmentation was observed by DNA gel electrophoresis; FCM showed that TB could affect the cell cycle progression of NB4 and MR2 cells simultaneously. , Induction of apoptosis, and the proportion of sub-G1 and G1 phase cells are similar; TUNEL assay also confirmed this result. In the process of inducing apoptosis, TB extended with time, and the expression level of bcl 2 gene gradually decreased. Conclusion TB has a dual role in synergistic retinoids-induced promyelocytic leukemia cell differentiation and apoptosis.