Matsuoka A, Miyamura K, Emi N, Tahara T, Tanimoto M, Naoe T, Ohno R, Kakizuka A, Evans R M, Saito H
First Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Leukemia. 1993 Aug;7(8):1151-5.
We have analyzed ten APL patients using reverse transcription polymerase chain reaction (RT-PCR) technique to detect PML/RAR alpha fused mRNA. All patients in this study had PML/RAR alpha fused mRNA (three cases of the short type and seven cases of the long type), although the chromosomal translocation t(15;17) was not detected in one patient. After ethidium bromide staining, two-thirds of the short type and all cases of the long type were found to have multiple PCR products (192 and 93 base pair (bp) bands in the short type and 666, 522, 263, and 164 bp in the long type). A total of six distinct fused mRNAs were sequenced (P1R1, P1R2, P3R1, P2R1, and P2R2). Southern hybridization analysis showed only one rearranged band in each of the patients. These results suggest that the longest mRNAs in each type are the authentic fused mRNAs and the other smaller mRNAs are generated through splicing events. In RAR alpha, a novel fusion point (R2) was identified within the fourth exon. This uncommon splicing may be caused by the instability of the splicing mechanism of the rearranged PML/RAR alpha gene. Among the ten APL patients, no correlation was observed between the type of fused mRNA and the clinical characteristics examined.
我们使用逆转录聚合酶链反应（RT-PCR）技术分析了10例急性早幼粒细胞白血病（APL）患者，以检测PML/RARα融合mRNA。本研究中的所有患者均有PML/RARα融合mRNA（3例短型和7例长型），尽管有1例患者未检测到染色体易位t(15;17)。溴化乙锭染色后，发现三分之二的短型和所有长型病例均有多个PCR产物（短型为192和93碱基对（bp）条带，长型为666、522、263和164 bp）。共对6种不同的融合mRNA进行了测序（P1R1、P1R2、P3R1、P2R1和P2R2）。Southern杂交分析显示，每位患者仅出现一条重排条带。这些结果表明，每种类型中最长的mRNA是真正的融合mRNA，其他较小的mRNA是通过剪接事件产生的。在RARα中，在第四外显子内鉴定出一个新的融合点（R2）。这种不常见的剪接可能是由重排的PML/RARα基因剪接机制的不稳定性引起的。在这10例APL患者中，未观察到融合mRNA类型与所检查的临床特征之间存在相关性。
