KLF4基因表达与JAK2 V617F点突变的关联性研究

Study on the relationship between KLF4 and JAK2 V617F point mutation

目的检测骨髓增殖性疾病(MPDs)中JAK2 V617F点突变与锌指蛋白转录因子(KLF4 mRNA)表达,探讨JAK2 V617F点突变与KLF4 mRNA表达水平与BCR/ABL阴性的MPDs的关系。方法用等位基因特异性聚合酶链反应(AS-PCR)技术检测4组JAK2 V617F点突变,根据结果分为阴性、阳性。反转录特异性聚合酶链反应(RT-PCR)技术检测4组KLF4 mRNA的表达水平。结果BCR/ABL阴性的MPDs的62例患者中JAK2 V617F基因突变慢性特发性骨髓纤维化(IMF)中有5例,阳性率55.56%(5/9),原发性血小板增多症(ET)中有15例,阳性率57.69%(15/26),真性红细胞增多症(PV)中有23例,阳性率88.46%(23/26),慢性嗜中性白血病1例。慢性粒细胞性白血病(CML)、急性白血病(AL)、对照组均没有阳性病例。KLF4mRNA在每例样本中均有表达。KLF4 mRNA表达水平在PV组显著高于其他3组(P<0.05)。结论BCR/ABL融合基因阴性的MPDs患者存在高频率JAK2 V617F点突变,KLF4 mRNA在PV患者中高表达。JAK2 V617F突变导致了KLF4基因表达的增高。JAK2 V617F突变阳性的ET患者,易出现并发症。AS-PCR是一种简单、有效检测JAK2 V617F突变方法。

Objective To detected JAK2V617F point mutation and the expression levels of the KLF4 mRNA of myeloproliferative diseases. To investigate the relationship between BCR/ABL negative MPDs and JAK2V617F point mutation as well as the expression levels of the KLF4 mRNA. Methods JAK2V617F point mutation was detected by allele-specific polymerase chain reaction. The expression levels of the KLF4 mRNA were detected by reverse transcription specific polymerase chain reaction. Results The JAK2V617F point mutation was detected in 62 BCR/ABL negative MPD patients, including 5/9 （55.56 % ） IMF patients, 15/26 （ 57.69 % ） ET patients, 23/26 （ 88.46 % ） PV patients and one CNL patient. While the somatic JAK2 V617F mutation was not found in either CML patients, the 12 AL patients or the 12 health adults. KLF4 mRNA expressed itself in everyone. The relative expression levels of KLF4 mRNA in PV group （mean value 1.0966, positive rate 100% ） were significantly higher than those of the other groups. Conclusion A somatic JAK2 V617F point mutation is found in a sizeable proportion of BCR/ABL fusion gene negative MPD patients. There is a significant difference in the expression levels of KLF4 mRNA between PV group and the other groups. JAK2 V617F mutation results in the increase of expression levels of KLF4 mRNA. JAK2 V617F positive ET patients have more complications, as compared with those of JAK2 V617F negative ET patients. AS-PCR is a sensitive and substantial technique in detecting JAK2 V617F mutation.