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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。
Objective To investigate the differentially expressed genes in HL-60 cells induced and not induced by all-trans retinoic acid. METHODS: Total RNA was extracted from human promyelocytic leukemia cell line, and cDNA was obtained by reverse transcription reaction (RT) mediated by anchored primer. PCR was carried out using the same anchored primer and another upstream primer as cDNA template, and PCR Amplification products were electrophoretically separated on a non-denaturing polyacrylamide gel and the gene expression profiles of the two groups of cells were compared. Results Differentially expressed HL-60 cells induced and not induced by all-trans retinoic acid were amplified by 24 primer combinations. A total of 541 fragments were amplified. The average number of bands per lane was 23, and 36 bands with significant differences were found. Conclusion mRNA differential display technique can effectively reflect the expression of multiple genes in HL-60 cells induced and not induced by all-trans retinoic acid.