論文白血病患者ETV基因異常檢測及臨床意義

1、ETV6論文：白血病患者ETV6基因異常檢測及臨床意義【中文摘要】1、檢測急慢性白血病病患者ETV6基因異常。2、探討ETV6基因異常在白血病的發(fā)病機(jī)制中的作用。3、初步揭示ETV6基因異常與白血病患者的預(yù)后的關(guān)系。材料與方法1、病例資料初診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,慢性淋巴細(xì)胞6例,全部符合2001年WHO的分型和診斷標(biāo)準(zhǔn)。男性14例,女性17例,中位年齡45歲(17歲-76歲)。2、骨髓細(xì)胞培養(yǎng)及染色體制備取患者骨髓標(biāo)本,在無刺激條件下培養(yǎng)并收獲細(xì)胞,采用直接法和(或)短期培養(yǎng)法制備細(xì)胞染色體。3、傳統(tǒng)細(xì)胞遺傳學(xué)采用R顯帶技術(shù)對患者骨髓細(xì)胞進(jìn)行染色

2、體分析。4、Split-signal FISH應(yīng)用分裂探針(RH77972和D12S922)對31例急慢性白血病患者染色體標(biāo)本進(jìn)行熒光原位雜交檢測其ETV6異常情況。5、FISH對檢測到ETV6缺失的患者用12號染色體著絲粒探針進(jìn)一步行FISH檢測12號染色體情況。6、RT-PCR對涉及ETV6異常的急性早幼粒細(xì)胞白血病患者行RT-PCR檢測PML-RARa融合基因。結(jié)果1、細(xì)胞遺傳學(xué)檢查31例患者發(fā)現(xiàn)12例存在染色體畸變,畸變率38.7%,其中涉及12號染色體畸變1例慢淋患者為12號三體,其它無涉及12號染色體異常。2、Split-signal FISH對31例白血病患者染色體標(biāo)本進(jìn)行熒光原

3、位雜交檢測其ETV6異常情況,在一例急性早幼粒細(xì)胞白血病(APL)患者檢測到ETV6缺失(部分或完全)。核型分析為正常核型。3、進(jìn)一步FISH用12號染色體著絲粒探針檢測該例存在ETV6缺失的APL患者的骨髓細(xì)胞均為2個信號。4、RT-PCR對該例存在ETV6缺失的患者進(jìn)行RT-PCR檢測：PML-RARa,凝膠電泳無產(chǎn)物,該患者PML-RARa融合基因陰性。結(jié)論1、該例存在ETV6缺失的APL患者PML-RARa陰性,ETV6的缺失與APL的發(fā)病密切相關(guān)。2、該例患者APL成功治療緩解后發(fā)生治療相關(guān)的MDS (t-MDS),伴有ETV6缺失的APL預(yù)后不良。3、國際首次報(bào)道伴有ETV6缺失P

4、ML-RARa融合基因陰性的APL患者,治療緩解后發(fā)生t-MDS.初步揭示APL的發(fā)生及其后來發(fā)生的t-MDS發(fā)病機(jī)制可能為ETV6的缺失使其抑癌活性喪失導(dǎo)致疾病發(fā)生。4、Split-signal FISH是檢測ETV6基因異常的準(zhǔn)確可靠的手段,與細(xì)胞遺傳學(xué)技術(shù)結(jié)合可以提高染色體異常的檢出率及準(zhǔn)確性?！居⑽恼縮To 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.【關(guān)鍵詞】ETV6 急性早幼粒細(xì)胞白血病 骨髓增生異常綜合癥 熒光原位雜交【英文關(guān)鍵詞】ETV6 Acute Promyelocytic Leukemia Myelodysplastic syndrome Fluorescence In situ hybridization【