期刊文献+

实时定量RT-PCR技术测定初治白血病患者常见融合基因转录子水平及其标准化的探讨 被引量:26

Detection of common fusion transcript levels in untreated leukemia patients by real-time quantitative RT-PCR technique
原文传递
导出
摘要 目的了解白血病常见融合基因 M-bcr-abl(表达产物为 P210^(bcr-abl))、m-bcr-abl(表达产物为 P190^(bcr-abl))、TEL-AML1、AML1-ETO、PML-RARα、CBFβ-MYH11在初治白血病患者中的表达水平。方法采用基于 TaqMan 探针的实时定量 RT-PCR(RQ-PCR)技术检测195例白血病患者骨髓细胞融合基因转录子水平,其中 M-bcr-abl^+慢性粒细胞白血病慢性期(CML-CP)50例、M-bcr-abl^+急性淋巴细胞白血病(ALL)10例、m-bcr-abl^+ ALL 19例、TEL-AML1^+ ALL 11例、AML1-ETO^+急性髓系白血病(AML)30例、PML-RARα^+急性早幼粒细胞白血病(APL)58例、CBFβ-MYH11^+ AML 患者17例,并检测13例 M-bcr-abl^+ CML-CP 患者的外周血细胞融合基因转录子水平。以 abl 作为内参基因,融合基因转录子水平以融合基因转录子拷贝数/abl 基因转录子拷贝数×100%表示。结果 CML-CP 患者骨髓及外周血 M-bcr-abl 转录子水平相似(中位值分别为30%和35%,P>0.05)、ALL 患者 M-bcr-abl 及m-bcr-abl 转录子水平相似(中位值分别为64%和54%,P>0.05),ALL 患者 M-bcr-abl 转录子水平明显高于 CML-CP 患者(P<0.001)。ALL 患者 TEL-AML1转录子中位水平为228%。表达特异性融合基因的 AML 患者中,AML1-ETO 转录子水平明显高于 CBFβ-MYH11及 PML-RARα(中位值分别为388%,145%和47%,P 值均<0.001),CBFβ-MYH11转录子明显高于 PML-RARα(P<0.001)。L 型、V 型及 S 型 PML-RARα转录子中位水平分别为45%、44%及55%,L 型明显低于 S 型(P=0.04)。结论不同类型融合基因转录子在初治白血病患者中的水平不同,并有个体差异。初治患者融合基因转录子水平的测定不仅为检测微量残留病、评价疗效提供基准值,而且是不同实验室间数据比较、实现标准化的基础。 Objective To evaluate levels of common specific fusion transcripts M-bcr-abl, m-bcr-abl,TEL-AML1, AML1-ETO, PML-RARα, CBFβ-MYHll in untreated leukemia patients. Methods Specific fusion transcript levels were detected by TaqMan-based real-time quantitative RT-PCR technique in a total of 208 samples, including 195 bone marrow samples from 50 M-bcr-abl( + ) chronic phase-chronic myeloid leukemia (CML-CP) , 10 M-bcr-abl( + ) acute lymphoblastic leukemia(ALL) , 19 m-bcr-abl( + ) ALL, 11 TEL-AML1( + ) ALL, 30 AML1-ETO( + ) acute myeloid leukemia(AML), 58 PML-RARα( + ) acute promyeloeytic leukemia(APL) and 17 CBFβ-MYH11 ( + ) AML patients and 13 peripheral blood samples from 13 M-ber-abl( + ) CML-CP patients, abl was chosen as internal control gene. Fusion transcript level was calculated as fusion transcript copies/abl transcript copies in percentage. Results Bone marrow and peripheral blood samples of CML-CP patients had similar M-bcr-abl fusion transcript levels ( median 30% vs 35%, P 〉 0.05 ). M- and m-bcr-abl ( median 64% vs 54% ) levels were similar in ALL patients ( P 〉 0.05 ), M-bcr-abl level was significantly higher in ALL than CML-CP patients( P 〈 0. 001 ). Median TEL-AML1 level was 228% in ALL patients. Among AML patients, AML1-ETO level was significantly higher than CBFβ- MYHll and PML-RARα levels (median 388% vs 145%, 388% vs 47%, all P 〈0.001 ), CBFβ-MYH11 level was significantly higher than PML-RARα level(P 〈 0. 001 ). Fusion transcript levels of L-, V- and S- type PML-RARα were 45% , 44% and 55% , respectively. L-type was significantly lower than S-type ( P =0.04). Conclusions Fusion transcript levels in untreated leukemia patients were different and patient-to-patient variations did exist. Detection of fusion transcript levels in untreated leukemia patients not only provides baseline for minimal residual disease monitoring and treatment evaluation but also enable the comparison in inter-laborato
出处 《中华血液学杂志》 CAS CSCD 北大核心 2007年第7期433-437,共5页 Chinese Journal of Hematology
关键词 白血病 融合蛋白质类 逆转录聚合酶链反应 实时定量 Leukemia Fusion proteins Reverse transciptase polymerase chain reaction, realtime, quantitative
  • 相关文献

参考文献11

  • 1Hughes T, Branford S. Molecular monitoring of bcr-abl as a guide to clinical management in chronic myeloid leukemia. Blood Rev, 2006, 20:29-41. 被引量:1
  • 2秦亚溱,刘艳荣,李金兰,王卉,常艳,付家瑜,阮国瑞,丘镜滢,陆道培,陈珊珊.慢性髓系白血病不同bcrabl融合基因转录子与临床关系的研究[J].中华血液学杂志,2003,24(7):347-350. 被引量:20
  • 3秦亚溱,李金兰,主鸿鹄,阮国瑞,李玲娣,张艳,许兰平,刘代红,黄晓军,陈珊珊,陆道培,刘艳荣.实时定量RT-PCR监测慢性粒细胞白血病患者造血干细胞移植后bcr-abl mRNA水平[J].中华血液学杂志,2006,27(8):511-514. 被引量:19
  • 4Beillard E, Pallisgaard N, van der Velden VH, et al. Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) -a Europe Against Cancer Program. Leukemia, 2003,17 : 2474-2456. 被引量:1
  • 5Gabert J, Beillard E, van der Velden VH,et al. Standardization and quality control studies of ' real-time ' quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia - a Europe Against Cancer program. Leukemia, 2003, 17:2318-2357. 被引量:1
  • 6Matin D, Kaeda J, Szydlo R, et al. Monitoring patients in complete cytogenetic remission after treatment of CML in chronic phase with imatinib : patterns of residual leukemia and prognostic factors for cytogenetic relapse. Leukemia, 2005, 19:507-512. 被引量:1
  • 7Asnafi V, Rubio MT, Delabesse E, et al. Prediction of relapse by day 100 bcr-abl quantification after allogeneic stem cell transplantation for chronic myeloid leukemia. Leukemia, 2006, 20:793-799. 被引量:1
  • 8Grimwade D, Lo Coco F. Acute promyelocytic leukemia: a model for the role of molecular diagnosis and residual disease monitoring in directing treatment approach in acute myeloid leukemia. Leukemia, 2002, 16 : 1959-1973. 被引量:1
  • 9Neumann F, Herold C, Hildebrandt B, et al. Quantitative real-time reverse-transcription polymerase chain reaction for diagnosis of bcr-abl positive leukemias and molecular monitoring following allogeneic stem cell transplantation. Eur J Haematol, 2003,70 : 1-10. 被引量:1
  • 10Branford S, Rudzki Z, Parkinson I, et al. Real-time quantitative PCR analysis can be used as a primary screen to identify patients with CML treated with imatinib who have bcr-abl kinase domain mutations. Blood, 2004, 104: 2926-2932. 被引量:1

二级参考文献21

  • 1Melo JV. The diversity of bcr-abl fusion proteins and their relationship to leukemia phenotype. Blood, 1996,88: 2375-2384. 被引量:1
  • 2Kwok S, Higuchi R . Avoiding false positives with PCR. Nature, 1989,339 : 237-238. 被引量:1
  • 3Pane F, lntrieri M, Quintarelli C, et al. BCR/ABL genes and leukemic phenotype: from molecular mechanisms to clinical correlations. Oncogene, 2002,21:8652-8667. 被引量:1
  • 4Perego RA, Costantini M, Comacchini G, et al. The possible influences of B2A2 and B3A2 BCR/ABL protein structure on thrombopoiesis in chronic myeloid leukaemia. Eur J Cancer, 2000,36 : 1395-1401. 被引量:1
  • 5Inokuchi K, Inoue T, Tojo A, et al. A possible correlation between the type of bcr-abl hybrid messenger RNA and platelet count in Philadelphiapositive chronic myelogenous leukemia Blood, 1991 , 78 : 3125-3127. 被引量:1
  • 6Saglio G, Pane F, Gottardi E, et al. Consistent amounts of acute leukemiaassociated P190^BCH/ABI transcripts are expressed by CML patients at diagnosis. Blood, 1996, 87:1075-1080. 被引量:1
  • 7van Rhee F, Hochhaus A, Lin F, et al. P190 BCR-ABL mRNA is expressed at low levels in p210-positive chronic myeloid and acute lymphoblastic leukemias. Blood, 1996, 87:5213-5217. 被引量:1
  • 8Hur M,Song EY, Kang SH, et al. Lymphoid preponderance and the absence of basophilia and splenomegaly are frequent in m-bcr-positive chronic myelogenous leukemia. Arm Hematol , 2002,81 : 219-223. 被引量:1
  • 9Yamaguchi H, lnokuchi K, Yokomizo E, et al. Philadelphia chromosomepositive acute myeloid leukemia with tetraploidy. In J Haematol, 2002,75 : 63-67. 被引量:1
  • 10Kelliher M, Knott A, Mclaughlin J, et al. Significance of the P210 versus P190 molecular abnormalities in adults with Philadelphia chromosomepositive acute leukemia. Mol Cell Biol, 1991,11:4710-4716. 被引量:1

共引文献36

同被引文献138

引证文献26

二级引证文献142

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部