经全反维甲酸诱导和未诱导的HL-60细胞基因的差异展示分析

mRNA Differential Display of Promyelocytic Leukemia Cell Lines (HL-60): Induced and Non-induced by All-trans Retinoid Acid

摘要:

目的 对经全反维甲酸诱导和未诱导的HL-60细胞差异表达基因进行初探。方法 提取早幼粒白血病细胞株中总RNA，经由锚定引物介导的逆转录反应(RT)获得cDNA；以cDNA为模板采用相同锚定引物和另一上游引物进行PCR扩增，然后将PCR扩增产物在非变性聚丙烯酰胺凝胶上进行电泳分离，比较两组细胞的基因表达情况。结果 经由24种引物组合对经全反维甲酸诱导和未诱导的HL-60细胞进行差异展示分析，扩增出541条片段，平均每泳道显带数23条，发现差异明显片段36条。结论 mRNA差异展示技术能有效地反映出经全反维甲酸诱导和未诱导的HL-60细胞多个基因差异表达情况。

Abstract:

Objective To study the differential display of promyletic leukemia cell lines induced and non-induced by all-trans ret inoid acid (ATRA). Methods cDNA fragment was obtained by revers e transcription-polymerse chain reaction (RT-PCR) with total RNA extracted from promyletic leukemia cell lines. The amplified cDNA fragment was confirmed on non -denaturing polyacrylamide gel. Results With 24 primer combinat ions, we amplified a total of 541 bands, the mean number of bands obtained per l ane was 23, and 36 bands were found with obvious differnce. Conclusions mRNA differential display is effective to detect and characterize alte red gene expressions in promyletic leukomia cell lines induced and non-induc ed by ATRA.