摘要
目的 利用逆转录 多重巢式聚合酶链反应 (reversetranscription Multiplexnestedpolymerasechainreaction ,RT MultiplexNestedPCR)分析 30例不同类型的白血病患者染色体畸变情况。方法 采用国内首次建立的一种RT MultiplexNestedPCR方法 ,同时筛选出 2 9种对急性白血病诊断和预后具有重要意义的染色体结构畸变。结果 14 / 19例白血病儿童携带有 7种染色体畸变或基因异常 ,包括t(1;19)、t(8;2 1)、dupMLL(11q2 3)、t(12 ;2 1)、TAL1D、HOX11和EVI1原癌基因活化 ;11/ 11例白血病成人中共检出 5种染色体易位或基因异常 ,包括t(8;2 1)、t(15 ;17)、t(9;2 2 )、HOX11和EVI1原癌基因活化。首次在 1例成人急性早幼粒细胞白血病 (acutepromyelocyticleukemia ,APL)标本中发现一种因早幼粒细胞白血病 (promyelocyticleukemia ,PML)基因剪接位置不同造成的新型PML/RARα融合转录本。结论 RT MultiplexNestedPCR可用于白血病患者初诊时染色体畸变的筛选 。
Objective The chromosomal aberration is one of the important lesions in leukemia It is difficult to get satisfied results with conventional chromosome karyotyping We have established an RT multiplex nested PCR to analyze leukemia samples and evaluate its clinical value Methods Thirty samples, which were from 19 pediatric acute leukemia patients (15 acute lymphoblastic leukemia, ALL, and 4 acute myelogenous leukemia, AML), aged from 1 5 to 16 years,and 11 adult leukemia patients (2 ALL, 5 AML, 1 hybrid acute leukemia, HAL, 2 chronic myelogenous leukemia, CML, and 1 chronic lymphoblastic leukemia, CLL), aged from 17 to 65 years were collected An RT multiplex nested PCR with the sequences involving 29 leukemia associated chromosome structural aberrations′ breakpoints (86 mRNA breakpoints or splice variants) was established This reaction, including 8 parallel PCR, could screen 29 chromosomal structural aberrations at the same time, which were important for diagnosis and prognosis of leukemia Results (1) Seventeen patients were detected with chromosome structural aberrations including t(1;19) (2 cases), t(8;21) (3 cases), t(9;22) (4 cases), t(12;21) (2 cases), t(15;17) (2 cases), dupMLL(11q23) (3 cases) and TAL1D (1 case) HOX11 was found activated in 7 cases of ALL and 1 case of AML Of these 8 patients, 6 patients also carried dupMLL, t(1;19) and t(9;22); another 2 patients were found to have activation of EVI1 The activation of EVI1 was involved in 13 lymphocytic leukemia, 1 AML, 1 CML and 1 HAL cases Ten out of 16 patients also had other chromosomal aberrations such as t(1;19), t(9;22), t(12;21), dupMLL or HOX11 activated (2) A new type of PML/RARA fused transcript in an adult APL patient who carried 2 types of PML/RARA fused transcripts was first reported One was L form as sequencing result indicated,the other fragment resulted from the different splicing site of PML gene The full length of the sequence belonged to the coding regions of PML and RARA genes, respectively There was no any identical sequence in the GeneBank So we thought that it was a new splice variant of t(15;17) Conclusions (1)We have established an RT multiplex nested PCR which, including 8 PCR, could screen 29 chromosome structural aberrations at the same time (2) Proto oncogene HOX11 could be activated in both ALL and AML, and always existed simultaneously with other chromosome structural aberrations or activation of EVI1 Proto oncogene EVI1 could be activated in ALL, AML, CML, CLL and HAL, and also showed up with other chromosome structural aberrations or activation of HOX11 (3) For the first time, we found a novel PML/RARA fused transcript in an APL patient, which resulted from a new splicing site of PML gene
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2001年第11期682-685,共4页
Chinese Journal of Pediatrics
