SYBR Green实时定量PCR检测急性髓系白血病患者BAALC基因表达

Detection of BAALC in patients with acute myeloid leukemia by real-time reverse transcriptase-polymerase chain reaction

目的：观察SYBR Green相对定量逆转录聚合酶链反应（RQ-PCR）方法检测急性髓系白血病（AML）患者外周血单个核细胞（PBMC）中脑和急性白血病细胞胞质（BAALC）基因mRNA的表达水平。方法：分离12例初发AML患者和5名健康成年人PBMC,以白血病细胞株K562为阳性对照,提取细胞总RNA并逆转录为cDNA,SYBR Green相对定量RQ-PCR检测BAALC基因表达水平。结果：健康成年人PBMC中检测不到BAALC mRNA的表达,12例AML患者中,有8例BAALC mRNA高表达,与K562比较,其相对值为K562的0.8~5.6倍。结论：用SYBR Green相对定量RQ-PCR可以检测到AML患者PBMC中BAALC基因的表达,该方法稳定、敏感、可靠。

Objective：To detect the mRNA expression of brain and acute leukemia,cytoplasmic（BAALC） in perpheral blood mononuclear cells（PBMC）,BAALC of patients with acute myeloid leukemia（AML） by comparative real-time reverse transcriptase-polymerase chain reaction（RQ-PCR） with SYBR Green.Methods：PBMC was obtained from 12 patients with AML and 5 healthy donors,from which total RNA was extracted and reverse transcribed into cDNA.The leukemia cell lines K562 cells was selected as positive control.The expression of BAALC was detected by RQ-PCR with SYBR Green assay.Results：The BAALC mRNA could not be detected in PBMC of healthy donors.However,in 12 patients with AML,it was highly expressed in 8 of them.The relative values were 0.8-5.6 compared with K562 cells.Conclusions：The expression of BAALC in PBMC of patients with AML could be detected by SYBR Green comparative RQ-PCR.It was a steady,sensitive and reliable assay method.