甲氨喋呤和三磷酸腺苷诱导U937细胞凋亡机制探讨  

INVESTIGATING THE MECHANISM OF U937 APOPTOSIS INDUCED BY METHOTREXATE AND ADENOSINE TRIPHOSPHATE

作　　者：卢青[1] 肖广芬[2] 陈艳华[2] 

机构地区：[1]浙江省杭州市第一人民医院,310006 [2]南华大学附二医院血液内科,衡阳421001

出　　处：《中国现代医学杂志》2002年第21期51-54,共4页China Journal of Modern Medicine

摘　　要：目的 :探讨甲氨喋吟、三磷酸腺苷诱导髓性白血病细胞系U937细胞凋亡过程中P73mRNA的表达变化 ,以进一步了解P73基因在U937细胞凋亡中的作用。方法 :用上述药物诱导U937细胞凋亡 ,凋亡指标采用细胞形态学、DNA片段电泳、流式细胞术检测DNA含量等方法 ;采用半定量逆转录PCR (RT -PCR)检测P73mRNA的表达变化。结果 :甲氨蝶呤、三磷酸腺苷均可诱导U937细胞凋亡。 5 μmol/L的甲氨蝶呤和0 .2 5g/L的三磷酸腺苷分别作用于U937细胞 6h、2 4h ,光镜下见细胞明显聚集、体积缩小 ,Wright’s +Giemsa染色见染色质明显浓缩、核碎裂等现象 ,而对照组未出现上述现象 ;DNA片断电泳见清晰的梯形条带 ;流式细胞仪检测 :5 μmol/L的甲氨蝶呤作用于U937细胞 6h、8h、10h ,细胞的凋亡率分别为 :5 .15 %、11.4 3%、14 .7% ,0 .2 5 g/L的三磷酸腺苷作用于U937细胞 2 4h、36h、4 8h细胞的凋亡率分别为 :2 .33%、11.90 %、35 .4 9% ,而对照组的细胞凋亡率分别为 :0 .0 0 %和 1.0 9% ;半定量RT -PCR结果 :与对照组相比 ,仅三磷酸腺苷处理 4 8h组P73mRNA的表达下调。结论 :在U937细胞凋亡过程中P73的mRNA表达差异无统计学意义。Objective:To determine the variation of p73 gene expression in the process of acute myeloid leukemia(AML) cell line U937 apoptosis induced by methotrexate(MTX) and adenosine triphosphate(ATP).Methods:Morphological changes of apoptosis were observed with invert microscope and Wright's+Giemsa staining. DNA Ladder and Sub-G1 Peak were examined by DNA gel electronphores and fluorescence flow cytometry (FCM) , respectively.We used semi-quantitive reverse transcription-polymerase chain reaction (RT-PCR) to examine the expression of p73 mRNA.Results:Both MTX and ATP can induce U937 cell apoptosis effectively. Condense of nuclear, fragmentations of chromosome and DNA Ladder were seen after 6h following treatment of 5μmol/L MTX and 24h following treatment of 0.25g/L ATP.Sub-G1 Peak were also determined by FCM, but the quantity of p73 expression was stable, except those cells which were treated with ATP for 48h.Conclusion:P73 mRNA level wasn't changed in U937 cell's apoptosis induced by MTX and ATP.

关 键 词：甲氨喋呤 三磷酸腺苷 细胞凋亡 U937细胞 髓性白血病 

分 类 号：R733.7[医药卫生—肿瘤] R965[医药卫生—临床医学]