p73基因在甲氨蝶呤诱导髓性白血病细胞系U937细胞凋亡过程中的表达  

p73 Gene Expression in Apoptotic Process of Acute Myeloid Leukemia Cell Line U937 Induced by Methotrexate

作　　者：肖广芬[1] 卢青[2] 杨向东[3] 

机构地区：[1]南华大学附属第二医院血液内科,衡阳421001 [2]杭州市第一人民医院,杭州310006 [3]南华大学心血管病研究所,衡阳421001

出　　处：《中国实验血液学杂志》2002年第2期104-107,共4页Journal of Experimental Hematology

摘　　要：为了探讨甲氨蝶呤 (MTX)诱导U937细胞凋亡过程中p73mRNA的表达变化 ,用MTX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含量及细胞周期的变化等方法 ;采用半定量反转录PCR(RT PCR)检测 p73mRNA的表达变化。研究结果显示 :MTX可诱导U937细胞凋亡。 5 μmol/L的MTX作用于U937细胞 6小时 ,可见部分细胞体积缩小 ,染色质明显浓缩和核碎裂。DNA片段电泳 ,MTX处理组可见清晰的梯形条带。流式细胞仪检测发现MTX作用于U937细胞 6 ,8,10小时 ,细胞的凋亡率分别为 5 .15 % ,11.4 3%和 14 .7% ,并使细胞阻滞在G2 /M +S期 ,而对照组细胞不发生凋亡。半定量RT PCR结果显示在U937细胞凋亡前后 ,p73mRNA的表达无明显变化。结论提示 ,在MTX诱导U937细胞凋亡过程中 。The purpose of this investigation was to study the variation of p73 gene expression in the apoptotic process of acute myeloid leukemia(AML) cell line U937 induced by methotrexate(MTX). Morphological changes of apoptotic cells were observed with microscopy and Wright′s+Giemsa staining. DNA ladder and cell cycle were examined by agarose gel electrophoresis and flow cytometry respectively. Using semi quantitive reverse transcription polymerase chain reaction (RT PCR), the expression of p73 mRNA was examined. Results showed that MTX could induce U937 cell apoptosis effectively. Condensed nuclei, fragmentation of chromosome and DNA ladder were seen after 6 hour following treatment of MTX 5 μmol/L. Sub G 1 peak and S+G 2/M arrest were also determined by FCM, but the quantity of p73 expression was generally constant. In conclusion, U937 cell apoptosis induced by MTX did not change p73 mRNA level.

关 键 词：P73基因 甲氨蝶呤 U937细胞 细胞凋亡 髓性白血病细胞系 基因表达 

分 类 号：R730.231[医药卫生—肿瘤] R733.7[医药卫生—临床医学]