荧光原位杂交技术可早期有效的预测慢性粒细胞性白血病的预后

doi:10.3969/j.issn.1673-8640.2015.02.016

摘要/Abstract

摘要： 目的

本研究探讨荧光原位杂交技术(FISH)在甲磺酸伊马替尼(IM)治疗慢性粒细胞性白血病(CML)长期预后中的早期预测价值。

方法

选取60例IM治疗阶段的CML患者骨髓标本,运用骨髓细胞形态学(BM)、FISH、逆转录-聚合酶链反应(RT-PCR)以及常规染色体分析方法(CC)对入组的CML患者每3~6个月进行48个月疗效观察。

结果

运用FISH技术在第3个月、第6个月以及第12个月检测融合基因Bcr-abl阳性细胞<10%的病例,其在第12个月具有较高的细胞遗传学完全缓解(CCR),同时,在第36个月和第48个月所观察的大多数患者均获得了无事件生存,与Bcr-abl阳性细胞百分数>10%的患者组比较,差异具有统计学意义(P<0.05)。相似的,运用RT-PCR和CC在第3个月和第6个月检测Bcr-Abl mRNA以及阳性分裂相百分数,两组病例在第36个月和第48个月的无事件生存率(EFS)差异无统计学意义(P>0.05),仅在第12个月差异具有统计学意义(P<0.05)。

结论

FISH分析技术可以为CML的长期预后提供早期有效的预测。

关键词: 慢性粒细胞性白血病, 荧光原位杂交技术, 无事件生存率, 早期预测价值

Abstract: Objective

To investigate the early predictive significance of fluorescence in-situ hybridization (FISH) in the long-term prognosis of treating chronic myeloid leukemia(CML) by imatinib(IM).

Methods

A total of 60 patients with CML undergoing IM treatment were enrolled, their bone marrow specimens were collected, and they were observed per 3-6 months for 48 months for treatment efficiency by bone marrow cell morphology(BM), FISH, reverse transcription polymerase chain reaction (RT-PCR) and karyotyping(CC).

Results

Applying FISH to detect the patients showed that BCR-ABL positive cell <10% in the 3rd month, the 6th month and the 12th month, which there was high CCR in the 12th month, and the most patients got free-event survival in the 36th and 48th months. Comparing with those whose BCR-ABL positive cell >10%, there was statistical significance (P<0.05). Similarly, applying RT-PCR and CC to detect Bcr-Abl mRNA and positive mitosis phase, there was no statistical difference in the 36th and 48th months(P>0.05), and there was statistical significance only in the 12th month (P<0.05).

Conclusions

FISH is able to provide the early and effective prediction for the long-term prognosis of CML.

Key words: Chronic myeloid leukemia, Fluorescence in-situ hybridization, Free-event survival rate, Early predictive significance

中图分类号:

Q503

彭友帆, 刘洋, 张朝霞. 荧光原位杂交技术可早期有效的预测慢性粒细胞性白血病的预后[J]. 检验医学, 2015, 30(2): 167-172.

PENG Youfan, LIU Yang, ZHANG Zhaoxia. Fluorescence in-situ hybridization can provide the early and effective prediction for the prognosis of chronic myeloid leukemia[J]. Laboratory Medicine, 2015, 30(2): 167-172.

图/表 6

参考文献 20

[1]	熊树民,余润泉.临床血液细胞学图谱与应用[M].上海:上海交通大学出版社,2009: 65.
[2]	HEHLMANN R, HOCHHAUS A, BACCARANI M.Chronic myeloid leukaemia[J]. Lancet, 2007, 370(9584): 342-350.
[3]	HOCHHAUS A, KANTARJIAN HM, BACEARANI M, et al.Dasatinib induces notable hematologic and cytogenetic responses in chronic-phase chronic myeloid leukemia after failure of imatinib therapy[J].Blood, 2007, 109(6): 2303-2309.
[4]	张之南,沈悌. 血液病诊断及疗效标准[M].3版.北京:科学出版社,2007: 134-138.
[5]	HANFSTEIN B, MÜLLER MC, HEHLMANN R, et al. Early molecular and cytogenetic response is predictive for long-term progression-free and overall survival in chronic myeloid leukemia (CML)[J].Leukemia, 2012, 26(9): 2096-2102.
[6]	TAKAHASHI N, KAMEOKA Y, TAGAWA H, et al.Early prediction of a long-term outcome by neutrophil-FISH in patients with CML receiving imatinib mesylate[J]. Int J Hematol, 2010, 92(3): 559-561.
[7]	TAKAHASHI N, MIURA I, KOBAYASHI Y, et al.Fluorescence in situ hybridization monitoring of BCR-ABL-positive neutrophils in chronic-phase chronic myeloid leukemia patients during the primary stage of imatinib mesylate therapy[J]. Int J Hematol, 2005, 81(3): 235-241.
[8]	OHM L, ARVIDSSON I, BARBANY G, et at. Early landmark analysis of imatinib treatment in CML chronic phase: less than 10% BCR-ABL by FISH at 3 months associated with improved long-term clinical outcome[J]. Am J Hematol, 2012, 87(8): 760-765.
[9]	The Italian Cooperative Study Group on Chronic Myeloid Leukemia. Long-term follow-up of the Italian trial of interferon-alpha versus conventional chemotherapy in chronic myeloid leukemia[J]. Blood, 1998, 92(5):1541-1548.
[10]	CUNEO A, BIGONI R, EMMANUEL B, et al.Fluorescence in situ hybridization for the detection and monitoring of the Ph-positive clone in chronic myelogenous leukemia: comparison with metaphase banding analysis[J]. Leukemia, 1998,12(11):1718-1723.
[11]	GRJMWADE D, BIONDI A, MOZZICONACCI M, et al.Characterization of acute promyelocytic leukemia cases lacking the classic t(15;17):results of the European Working Party[J].Blood,2000, 96(4):1297-1308.
[12]	王小青,唐少华,裘晓乐.荧光原位杂交技术检测BCR/ABL融合基因[J].检验医学, 2006,21(2):152-154.
[13]	KANTARJIAN HM, TALPAZ M, CORTES J, et al.Quantitative polymerase chain reaction monitoring of BCR-ABL during therapy with imatinib mesylate (STI571; gleevec) in chronic-phase chronic myelogenous leukemia[J]. Clin Cancer Res, 2003, 9(1):160-166.
[14]	BRANFORD S, HUGHES TP, RUDZKI Z.Monitoring chronic myeloid leukaemia therapy by real-time quantitative PCR in blood is a reliable alternative to bone marrow cytogenetics[J]. Br J Haematol, 1999,107(3):587-599.
[15]	HOCHHAUS A, REITER A, SAUSSELE S, et al.Molecular heterogeneity in complete cytogenetic responders after interferon-alpha therapy for chronic myelogenous leukemia: low levels of minimal residual disease are associated with continuing remission. German CML Study Group and the UK MRC CML Study Group[J]. Blood, 2000, 95(1):62-66.
[16]	HUGHES TP, HOCHHAUS A, BRANFORD S, et al.Long-term prognostic significance of early molecular response to imatinib in newly diagnosed chronic myeloid leukemia: an analysis from the International Randomized Study of Interferon and STI571 (IRIS)[J]. Blood, 2010,116(19):3758-3765.
[17]	KLAMOVÁ H, POLÁKOVÁ KM, MUŽIK J. Evaluation of 5-year imatinib treatment of 458 patients with CP-CML in routine clinical practice and prognostic impact of different BCR-ABL cutoff levels[J]. Cancer Med, 2013, 2(2):216-225.
[18]	ZHEN C, WANG YL.Molecular monitoring of chronic myeloid leukemia international standardization of BCR-ABL 1 quantitation[J]. J Mol Diagn, 2013,15(5):556-564.
[19]	BRANFORD S, FLETCHER L, CROSS NC, et al.Desirable performance characteristics for BCR-ABL measurement on an international reporting scale to allow consistent interpretation of individual patient response and comparison of response rates between clinical trials[J]. Blood,2008,112(8):3330-3338.
[20]	秦亚溱,主鸿鸽.慢性髓性白血病分子监测手册[M].北京:人民卫生出版社,2013:62-66.

缓解率	3月	6月	9月	12月	24月	36月	48月
CHR	52(79)	39(97)	46(99)	53(96)	51(100)	46(100)	28(100)
CCR	49(18)	45(54)	39(67)	51(80)	39(97)	35(98)	28(96)
MMR	45(4)	46(11)	46(32)	49(41)	51(73)	38(71)	25(84)

缓解率	3月	6月	9月	12月	24月	36月	48月
CHR	52(79)	39(97)	46(99)	53(96)	51(100)	46(100)	28(100)
CCR	49(18)	45(54)	39(67)	51(80)	39(97)	35(98)	28(96)
MMR	45(4)	46(11)	46(32)	49(41)	51(73)	38(71)	25(84)

检测方法	阳性 (%)	第3个月检测			第6个月检测			第12个月检测
检测方法	阳性 (%)	CCR (12月)	EFS (36月)	EFS (48月)	CCR (12月)	EFS (36月)	EFS (48月)	EFS (36月)	EFS (48月)
CC	<10	25/26	25/25	26/26	20/22	18/20	18/20	35/36	34/35
	>10	14/21	15/18	15/18	14/23	16/23	15/22	7/15	7/15
	P值	0.022	0.131	0.122	0.046	0.270	0.179	0.000	0.000
FISH	<10	28/30	29/30	29/30	32/37	23/30	23/30	23/25	23/25
	>10	14/22	14/23	13/23	2/8	2/7	2/7	1/5	1/5
	P值	0.020	0.003	0.002	0.001	0.001	0.046	0.002	0.002
PCR	<10	16/18	18/20	17/19	30/35	27/30	24/27	37/42	34/38
	>10	13/18	12/18	11/17	7/11	11/15	11/15	1/4	1/4
	P值	0.400	0.173	0.144	0.240	0.388	0.309	0.000	0.004

检测方法	阳性 (%)	第3个月检测			第6个月检测			第12个月检测
检测方法	阳性 (%)	CCR (12月)	EFS (36月)	EFS (48月)	CCR (12月)	EFS (36月)	EFS (48月)	EFS (36月)	EFS (48月)
CC	<10	25/26	25/25	26/26	20/22	18/20	18/20	35/36	34/35
	>10	14/21	15/18	15/18	14/23	16/23	15/22	7/15	7/15
	P值	0.022	0.131	0.122	0.046	0.270	0.179	0.000	0.000
FISH	<10	28/30	29/30	29/30	32/37	23/30	23/30	23/25	23/25
	>10	14/22	14/23	13/23	2/8	2/7	2/7	1/5	1/5
	P值	0.020	0.003	0.002	0.001	0.001	0.046	0.002	0.002
PCR	<10	16/18	18/20	17/19	30/35	27/30	24/27	37/42	34/38
	>10	13/18	12/18	11/17	7/11	11/15	11/15	1/4	1/4
	P值	0.400	0.173	0.144	0.240	0.388	0.309	0.000	0.004