二次PCR检测HL_（60）细胞系PML－RAR_α融合mRNA

Detection of PML-RAR_α fusion mRNA in HL_(60) cell line by booster-PCR

急性早幼粒细胞白血病（ＡＰＬ）具有染色体易位ｔ（１５；１７）而产生的特异的融合基因ＰＭＬ－ＲＡＲ_α。本研究应用二次聚合酶链反应（ｂｏｏｓｔｅｒＰＣＲ）技术检测了ＡＰＬ细胞系ＨＬ_（６０）细胞中ＰＭＬ－ＲＡＲ_α融合ｍＲＮＡ。结果表明，ＨＬ_（６０）细胞系中同时具有二种ＰＭＬ－ＲＡＲ_α融合ｍＲＮＡ异构体（Ｌ型和Ｓ型）。本实验的敏感度可达１０￣（－６）个细胞水平，同时检测了结果具有高度的特异性。提示本方法可作为今后检测ＡＰＬ微量残留白血病细胞一种灵敏、有效的手段。

It is shown that the t (15;17) translocation specifically associates with acute promyelocytic leukemia(APL) and forms the PML-RAR_α fusion gene. Now, we detect the PML-RAR_α fusion mRNA in APL-derived cell line HL_(60) by booster polymerase chain reaction. Our results show that there are two PML-RAR_α fusion mRNA isoforms (p6-r3 and p3-r3) in HL_(60)cell line.It demonstrates that PML-RAR_α fusion gene is alloplasmatic further. We also obtain the proliferating fragments as the same length as expected when detecting the fusion gene by using HeminestedPCR.But,we did not get any proliferation products in CEM cell line as a nagative control.The sensitivity of detection get 10￣(-6) by our booster-PCR technique.Our results suggest that the technique is a sensitive method for the assessment of minimal residual disease in APL.