论文白血病患者ETV基因异常检测及临床意义

1、ETV6论文：白血病患者ETV6基因异常检测及临床意义【中文摘要】1、检测急慢性白血病病患者ETV6基因异常。2、探讨ETV6基因异常在白血病的发病机制中的作用。3、初步揭示ETV6基因异常与白血病患者的预后的关系。材料与方法1、病例资料初诊的急慢性白血病患者31例,其中急性白血病13例,慢性粒白血病12例,慢性淋巴细胞6例,全部符合2001年WHO的分型和诊断标准。男性14例,女性17例,中位年龄45岁(17岁-76岁)。2、骨髓细胞培养及染色体制备取患者骨髓标本,在无刺激条件下培养并收获细胞,采用直接法和(或)短期培养法制备细胞染色体。3、传统细胞遗传学采用R显带技术对患者骨髓细胞进行染色

2、体分析。4、Split-signal FISH应用分裂探针(RH77972和D12S922)对31例急慢性白血病患者染色体标本进行荧光原位杂交检测其ETV6异常情况。5、FISH对检测到ETV6缺失的患者用12号染色体着丝粒探针进一步行FISH检测12号染色体情况。6、RT-PCR对涉及ETV6异常的急性早幼粒细胞白血病患者行RT-PCR检测PML-RARa融合基因。结果1、细胞遗传学检查31例患者发现12例存在染色体畸变,畸变率38.7%,其中涉及12号染色体畸变1例慢淋患者为12号三体,其它无涉及12号染色体异常。2、Split-signal FISH对31例白血病患者染色体标本进行荧光原

3、位杂交检测其ETV6异常情况,在一例急性早幼粒细胞白血病(APL)患者检测到ETV6缺失(部分或完全)。核型分析为正常核型。3、进一步FISH用12号染色体着丝粒探针检测该例存在ETV6缺失的APL患者的骨髓细胞均为2个信号。4、RT-PCR对该例存在ETV6缺失的患者进行RT-PCR检测：PML-RARa,凝胶电泳无产物,该患者PML-RARa融合基因阴性。结论1、该例存在ETV6缺失的APL患者PML-RARa阴性,ETV6的缺失与APL的发病密切相关。2、该例患者APL成功治疗缓解后发生治疗相关的MDS (t-MDS),伴有ETV6缺失的APL预后不良。3、国际首次报道伴有ETV6缺失P

4、ML-RARa融合基因阴性的APL患者,治疗缓解后发生t-MDS.初步揭示APL的发生及其后来发生的t-MDS发病机制可能为ETV6的缺失使其抑癌活性丧失导致疾病发生。4、Split-signal FISH是检测ETV6基因异常的准确可靠的手段,与细胞遗传学技术结合可以提高染色体异常的检出率及准确性。【英文摘要】sTo explore the abnormality of ETV6 gene in patients with acute leukemia or chronic leukemia.To explore the potential oncogenic mechanisms of E

5、TV6 gene abnormality in leukemia.To announce initially the relationship of ETV6 gene abnormality and prognosis of leukemia.Materials andMethodsThirty-one newly diagnosed patients (14 males,17 females, mean age 45 years, range 17-76 years)were studied. The diagnosis of acute leukemia or chronic leuke

6、mia was based on 2001 WHO classification and thediagnostic criteria.Cytogenetic analysis of bone marrow cells was performed by direct method and/or 24 h culture method. Chromosome abnormality by conventional cytogenetics of R-banding technique.Split-FISH analysis with split probes RH77972 and D12S92

7、2 was performed on the bone marrow samples to detect the abnormality of ETV6.Further FISH experiment was performed using chromosome 12 centromere probe in APL patient with part deletion of ETV6 gene.PML/RARfusion gene was detected by RT-PCR in APL patient with part deletion of ETV6 gene.ResultsBone

8、marrow cells R-banding chromosome karyotype analysis of 31 cases of luekemia patients showed 12 of 31 had chromosome abnormalies, of which 1 case with +12, and the traditional cytogenetic analysis had not found any other chromosome abnormalies of chromosome 12.A APL patient with part deletion of ETV

9、6 was detected by Split-signal FISH, with normal karyotype 46 XX.Further FISH showed two signal in every bone marrow cell.RT-PCR did not detect PML/RARa fusion gene products in sample of this APL patient with part or whole deletion of ETV6.ConclusionsPart deletion of ETV6 was detected in the APL,but

10、 this APL was one case without PML/RARa fusion gene, suggesting that it was close relationship between the pathogenesis of APL and the deletion of ETV6.This case APL without PML-RARa but with part deletion of ETV6 was prognosised Myelodysplastic syndrome MDS afert successful APL therapy, and has a b

11、ad prognosis. To our knowledge, this is the first time to report t-MDS of APL only with deletion of ETV6 after successful APL therapy. The deletion of ETV6 lead to the ETV6 protein inactive and losing the function of inhibiting tumor, so the myeloid hematopoietic cells proliferate excessively and pr

12、ocess to hematological malignancies.Split-signal FISH is a reliable measure to detect abnormality of ETV6 gene. Combination of the cytogenetic and FISH analysis can bring about more comprehensive and accurate results.【关键词】ETV6 急性早幼粒细胞白血病 骨髓增生异常综合症 荧光原位杂交【英文关键词】ETV6 Acute Promyelocytic Leukemia Myelodysplastic syndrome Fluorescence In situ hybridization【