RT-PCR结合RFL P技术分析胰岛素样生长因子-2基因的印迹丢失  

Detection of Loss of Imprinting of the Insulin-Like Growth Factor Ⅱ Gene

作　　者：杨本海[1] 刘永忠[1] 

机构地区：[1]皖南医学院附属弋矶山医院临床医学研究所,241001

出　　处：《皖南医学院学报》2001年第3期169-171,共3页Journal of Wannan Medical College

摘　　要：目的　建立逆转录聚合酶链反应 (RT PCR)结合限制性片段长度多态性 (RFLP)技术 ,分析急、慢性髓性白血病的胰岛素样生长因子 2 (IGF 2 )基因的印迹丢失 (Lossofimprinting ,LOI)。 方法　利用IGF 2基因第 9外显子具有ApaⅠ酶多态性 ,首先进行PCR RFLP筛选杂合子信息个体 ;RT PCR RFLP结合银染技术分析杂合子信息个体IGF 2基因的LOI。结果　 2 6例AML患者中 11例为杂合子信息个体 ;其中 8例为双等位基因表达 ,即发生了LOI ;35例CML患者中 18例为杂合子信息个体 ,其中 8例为双等位基因表达。被检患者IGF 2基因第 9外显子ApaⅠ位点杂合子频率为 47 5 %(2 9/ 6 1)。结论　RT PCR RFLP能简便、快速、准确地检测IGF 2基因的LOI。Objective To develop a simple and rapid method of reverse transcription polymerase chain reaction(RT-PCR) and restriction fragment length polymorphism (RFLP) to analyze loss of imprinting(LOI) of insulin-like growth factor Ⅱ (IGF-2) gene in acute myeloid leukemia(AML) and chronic myeloid leukemia(CML).Methods By using the polymorphism site in 9 exon with Apa Ⅰ in IGF-2 gene, heterozygous patients were selected by PCR-RFLP and frequency of LOI of IGF-2 gene was further examined with RT-PCR-RFLP technique.Results 11 AMl heterozygous cases for IGF-2 gene were selected from among 26 AML patients and 8 cases were further detected as loss of imprinting of IGF-2 of 18 heterozygous cases from 35 CML patients, 8 cases were further detected as loss of imprinting of IGF-2.Of the patients detected, the frequency of heterozygote for Apa Ⅰ site was 47.5%(29/61).Conclusion The method used in this study is simple, rapid and accurate for detecting LOI of IGF-2.

关 键 词：胰岛素生长因子-2 印迹丢失 逆转录聚合酶链反应 限制性片段长度多态性 急性 慢性 髓性白血病 

分 类 号：R733.7[医药卫生—肿瘤] Q754[医药卫生—临床医学]