实时荧光定量聚合酶链反应法监测慢性髓系白血病外周血BCR-ABL融合基因水平的价值研究
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摘要
目的探讨实时荧光定量聚合酶链反应(qRT-PCR)法监测慢性髓系白血病(CML)外周血BCR-ABL融合基因水平的价值。方法选取2016年3月～2017年6月我院血液科收治的CML患者35例,应用qRT-PCR法检测患者的BCR-ABL融合基因水平,并监测不同疾病分期患者BCR-ABL转录水平,动态监测不同BCR-ABL融合基因水平组患者伊马替尼的治疗效果及无病生存期(DFS),患者治疗前后的BCR-ABL转录水平变化。结果急变期患者的BCR-ABL转录本水平明显高于加速期和慢性期,加速期患者的BCR-ABL转录本水平明显高于慢性期,组间两两比较差异均有统计学意义(F=4.68,P=0.00)。疗效满意方面:低水平组与中水平组患者比较,差异无统计学意义(P>0.05),但两组均明显高于高水平组,差异有统计学意义(P<0.05);中位DFS方面:BCR-ABL融合基因低水平组患者的中位DFS明显长于中水平组、高水平组,中水平组的中位DFS明显长于高水平组,组间两两比较差异均有统计学意义(F=36.15,P=0.00)。伊马替尼靶向治疗第3、6、12个月后的BCR-ABL转录本水平均明显低于治疗前(P=0.00);通过伊马替尼靶向治疗,患者的BCR-ABL转录本平明显降低,但前6个月的下降幅度最明显。结论 qRT-PCR技术可准确地监测患者外周血BCR-ABL融合基因水平,有利于辅助CML患者疾病的诊断和病情分期,同时,通过对CML患者不同时期外周血BCR-ABL融合基因水平的动态监测有利于准确评估患者的治疗效果和预后,对CML患者治疗方案的选择具有重要指导意义。
Objective To investigate the value of quantitative real-time polymerase chain reaction(qRT-PCR) method monitoring the BCR-ABL fusion gene level in peripheral blood of chronic myeloid leukemia(CML). Methods A total of 35 patients with CML treated in our department of hematology from March 2016 to June 2017 were selected. The BCRABL fusion gene level was detected by qRT-PCR in patients. The transcriptional level of BCR-ABL in patients with different disease stages were monitored. The therapeutic effect and disease-free survival(DFS) of Imatinib in patients with different BCR-ABL fusion gene level were monitored dynamically. The transcriptional levels of BCR-ABL in patients before and after treatment were also monitored. Results The transcriptional level of BCR-ABL in patients with acute phase was significantly higher than that in patients with accelerated phase and chronic phase. The transcriptional level of BCR-ABL in patients with accelerated phase was significantly higher than that in patients with chronic phase,and there were significant differences between the two groups(F=4.68, P=0.00). In terms of curative effect satisfaction,there was no significant difference between the low-level group and the middle-level group(P >0.05), but the two groups were significantly higher than the high-level group(P<0.05). In terms of median DFS, the middle DFS of BCRABL fusion gene in the low-level group was significantly longer than that of the middle-level group and the high-level group, the median DFS in the middle-level group was significantly longer than that in the high-level group, and there were significant differences between the two groups(F=36.15, P=0.00). BCR-ABL transcripts level after Imatinib targeted therapy at 3, 6 and 12 months were significantly lower than those before treatment(P=0.00). BCR-ABL transcripts level were significantly lower in patients with Imatinib targeted therapy, but the most significant decrease was in the first six months. Conclusion q RT-PCR technology can accurately monitor the level of BCR-ABL fusion gene in peripheral blood of patients, which is helpful to assist the diagnosis and stage of disease in patients with CML. At the same time, dynamic monitoring BCR-ABL fusion gene level in peripheral blood of CML patients at different stages is helpful to accurately evaluate the therapeutic effect and prognosis of patients, and has important guiding significance for the selection of treatment options for patients with CML.
Objective To investigate the value of quantitative real-time polymerase chain reaction(qRT-PCR) method monitoring the BCR-ABL fusion gene level in peripheral blood of chronic myeloid leukemia(CML). Methods A total of 35 patients with CML treated in our department of hematology from March 2016 to June 2017 were selected. The BCRABL fusion gene level was detected by qRT-PCR in patients. The transcriptional level of BCR-ABL in patients with different disease stages were monitored. The therapeutic effect and disease-free survival(DFS) of Imatinib in patients with different BCR-ABL fusion gene level were monitored dynamically. The transcriptional levels of BCR-ABL in patients before and after treatment were also monitored. Results The transcriptional level of BCR-ABL in patients with acute phase was significantly higher than that in patients with accelerated phase and chronic phase. The transcriptional level of BCR-ABL in patients with accelerated phase was significantly higher than that in patients with chronic phase,and there were significant differences between the two groups(F=4.68, P=0.00). In terms of curative effect satisfaction,there was no significant difference between the low-level group and the middle-level group(P >0.05), but the two groups were significantly higher than the high-level group(P<0.05). In terms of median DFS, the middle DFS of BCRABL fusion gene in the low-level group was significantly longer than that of the middle-level group and the high-level group, the median DFS in the middle-level group was significantly longer than that in the high-level group, and there were significant differences between the two groups(F=36.15, P=0.00). BCR-ABL transcripts level after Imatinib targeted therapy at 3, 6 and 12 months were significantly lower than those before treatment(P=0.00). BCR-ABL transcripts level were significantly lower in patients with Imatinib targeted therapy, but the most significant decrease was in the first six months. Conclusion q RT-PCR technology can accurately monitor the level of BCR-ABL fusion gene in peripheral blood of patients, which is helpful to assist the diagnosis and stage of disease in patients with CML. At the same time, dynamic monitoring BCR-ABL fusion gene level in peripheral blood of CML patients at different stages is helpful to accurately evaluate the therapeutic effect and prognosis of patients, and has important guiding significance for the selection of treatment options for patients with CML.
引文
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[11]吴薇,韩瑞玲,张力,等.RQ-PCR在慢性粒细胞白血病细胞BCR-ABL融合基因检测中的应用[J].检验医学与临床,2016,13(22):3133-3134.
[12]Shamsipur M,Nasirian V,Barati A,et al.Determination o c DNA encoding BCR/ABL fusion gene in patients with chronic myelogenous leukemia using a novel FRET-based quantum dots-DNA nanosensor[J].Anal Chim Acta,2017,19(6):62
[13]程静,陈少谦,江晓冰,等.BCR/ABL融合基因阳性的急性淋巴细胞白血病的细胞形态与免疫表型特点[J].实用医学杂志,2016,32(24):4125-4128.
[14]Martin-Cabrera P,Haferlach C,Kern W,et al.BCR-ABL1-positive and JAK2 V617F-positive clones in 23 patients with both aberrations reveal biologic and clinical importance[J].Br J Haematol,2016,176(1):283-288.
[15]任海英,戚行江,梁森苗,等.利用常规PCR和实时荧光定量PCR检测杨梅凋萎病菌[J].植物病理学报,2016,46(1):1-10.
[16]Diamond JMS,Almeida AMD.CALR-mutated primary myel ofibrosis evolving to chronic myeloid leukemia with both CALR mutation and BCR-ABL1 fusion gene[J].Ann Hematol,2016,95(12):1-4.
[17]董珊珊,郭英,谭红丽,等.应用实时荧光定量PCR建立鼠疫特异基因诊断技术[J].中华地方病学杂志,2016,35(2):119-122.
[18]Wu J,Huang Y,Bian X,et al.Biosensing of BCR/ABLfusion gene using an intensity-interrogation surface plasmon resonance imaging system[J].Opt Commun,2016,377:24-32.
[19]Chuang TJ,Wu CS,Chen CY,et al.NCLscan:accurate identification of non-co-linear transcripts(fusion,trans-splicing and circular RNA)with a good balance between sensitivity and precision[J].Nucleic Acids Res,2016,44(3):e29.
[20]肖恒,任彦斌,杨志明,等.实时荧光定量PCR检测BCR-ABL融合基因的临床意义[J].中国优生与遗传杂志,2017(5):22-23.
[2]Kurita D,Hatta Y,Hojo A,et al.Adult acute lymphoblastic leukemia with a rare b3a3 type BCR/ABL1 fusion transcript[J].Cancer Genet,2016,209(4):161-165.
[3]陈曦希,杨明珍,夏瑞祥.调节性T细胞及相关细胞因子在慢性髓系白血病中的变化[J].安徽医科大学学报,2016,51(7):1015-1018.
[4]Tong YQ,Zhao ZJ,Liu B,et al.New rapid method to detect BCR-ABL fusion genes with multiplex RT-qPCR in onetube at a time[J].Leuk Res,2018,1(8):69.
[5]张江召,赵哲,黎纬明,等.国产伊马替尼与原研伊马替尼治疗初诊慢性髓性白血病慢性期的临床疗效及用药安全性对比分析[J].临床血液学杂志,2017,5(4):527-531.
[6]Duan MH,Li H,Cai H.A rare e13a3(b2a3)BCR-ABL1 fusion transcript with normal karyotype in chronic myeloid leukemia:The challenges in diagnosis and monitoring minimal residual disease(MRD)[J].Leuk Res,2017,8(22):8.
[7]徐俊卿,徐泽锋,王静雅,等.615例Ph染色体/BCR-ABL融合基因阴性骨髓增殖性肿瘤患者的症状负荷评估[J]中华血液学杂志,2016,37(1):26-29.
[8]许大兵,孙余婕,沈佐君.CML患者TGF-β1mRNA与bcr ablP210融合基因转录本表达的关系[J].检验医学,2016,31(8):671-674.
[9]Manthawornsiri Y,Polpanich D,Yamkamon V,et al.Magnetic Nanoparticles PCR Enzyme-Linked Gene Assay for Quantitative Detection of BCR/ABL Fusion Gene in Chronic Myelogenous Leukemia[J].J Clin Lab Anal,2016,30(5):534-542.
[10]庞丽,权文强,吴军录,等.TaqMan-MGB实时荧光定量PCR检测RIG-G基因方法的建立及其方法学验证[J]中华检验医学杂志,2016,39(12):936-935.
[11]吴薇,韩瑞玲,张力,等.RQ-PCR在慢性粒细胞白血病细胞BCR-ABL融合基因检测中的应用[J].检验医学与临床,2016,13(22):3133-3134.
[12]Shamsipur M,Nasirian V,Barati A,et al.Determination o c DNA encoding BCR/ABL fusion gene in patients with chronic myelogenous leukemia using a novel FRET-based quantum dots-DNA nanosensor[J].Anal Chim Acta,2017,19(6):62
[13]程静,陈少谦,江晓冰,等.BCR/ABL融合基因阳性的急性淋巴细胞白血病的细胞形态与免疫表型特点[J].实用医学杂志,2016,32(24):4125-4128.
[14]Martin-Cabrera P,Haferlach C,Kern W,et al.BCR-ABL1-positive and JAK2 V617F-positive clones in 23 patients with both aberrations reveal biologic and clinical importance[J].Br J Haematol,2016,176(1):283-288.
[15]任海英,戚行江,梁森苗,等.利用常规PCR和实时荧光定量PCR检测杨梅凋萎病菌[J].植物病理学报,2016,46(1):1-10.
[16]Diamond JMS,Almeida AMD.CALR-mutated primary myel ofibrosis evolving to chronic myeloid leukemia with both CALR mutation and BCR-ABL1 fusion gene[J].Ann Hematol,2016,95(12):1-4.
[17]董珊珊,郭英,谭红丽,等.应用实时荧光定量PCR建立鼠疫特异基因诊断技术[J].中华地方病学杂志,2016,35(2):119-122.
[18]Wu J,Huang Y,Bian X,et al.Biosensing of BCR/ABLfusion gene using an intensity-interrogation surface plasmon resonance imaging system[J].Opt Commun,2016,377:24-32.
[19]Chuang TJ,Wu CS,Chen CY,et al.NCLscan:accurate identification of non-co-linear transcripts(fusion,trans-splicing and circular RNA)with a good balance between sensitivity and precision[J].Nucleic Acids Res,2016,44(3):e29.
[20]肖恒,任彦斌,杨志明,等.实时荧光定量PCR检测BCR-ABL融合基因的临床意义[J].中国优生与遗传杂志,2017(5):22-23.