实时荧光定量PCR法检测外周血中BCR/ABL基因表达在Ph+急性淋巴细胞白血病患者造血干细胞移植治疗中的应用

doi:10.13481/j.1671-587X.20220122

摘要/Abstract

摘要： 目的

采用实时荧光定量PCR（RT-qPCR）法定期检测费城染色体阳性急性淋巴细胞白血病（Ph⁺ALL）患者外周血中BCR/ABL融合基因表达的动态变化，为患者制订不同的治疗方案，以提高患者总生存（OS）率。

方法

回顾性分析20例初治和难治复发Ph⁺ALL患者的临床资料，经过酪氨酸激酶抑制剂（TKI）+长春新碱+强的松（TKI+VP）或者酪氨酸激酶抑制剂+长春新碱+柔红霉素+强的松（TKI+VDP）等多种化疗方案诱导治疗，17例患者达到血液学完全缓解（HCR），3例未缓解（NR）。其中HCR患者中8例患者微小残留病（MRD）为阴性，3例患者选择自体造血干细胞移植（Auto-HSCT），在造血功能重建后均接受了TKI维持治疗至今；另5例MRD阴性患者、9例MRD阳性患者和3例NR患者均进行了异基因造血干细胞移植（Allo-HSCT）。所有患者分别在干细胞移植前和移植后1、2、3、6、9及12个月进行外周血BCR/ABL融合基因检测，评估患者的MRD。Allo-HSCT组患者造血重建后，均给予TKI口服干预治疗，其后基因检测持续阴性者，TKI口服干预治疗1年后停用。治疗期间如果监测MRD转阳则依据移植时间给予更换有效TKI药物或减停免疫抑制剂等治疗措施。

结果

全组患者造血干细胞移植后均达到造血重建，粒细胞植活中位时间为14（11~24）d，血小板植活中位时间为17（13~52）d。随访至2020年12月，中位随访时间为52（3~111）个月。全组患者3年OS率为（61.1±11.7）%。Allo-HSCT组和Auto-HSCT组患者移植后3年OS率分别为（52.8±13.4）%和100.0%，组间比较差异无统计学意义（P=0.178）。移植前MRD阴性组和移植前HCR但MRD阳性组患者3年OS率分别为（87.5±11.7）%和（42.8±15.6）%，组间比较差异无统计学意义（P=0.065）。移植前NR的患者全部死亡。全组死于疾病复发5例，死于肺感染1例，死于Ⅳ度急性移植物抗宿主病（GVHD）1例。

结论

对Ph⁺ALL患者依据BCR/ABL融合基因检测结果的动态变化选择Auto-HSCT或Allo-HSCT及术后分层干预治疗措施，可提高患者的OS率。

关键词: 费城染色体阳性急性淋巴细胞白血病, 造血干细胞移植, 实时荧光定量聚合酶链反应, 融合基因

Abstract: Objective

To examine the dynamic changes in BCR/ABL fusion gene expression in peripheral blood in the patients with Ph⁺ acute lymphoblastic leukemia （Ph⁺ALL） by real-time fluorescence quantitative PCR（RT-qPCR），and to formulate the different treatment measures for different patients and improve the overall survival（OS） rates of patients.

Methods

The clinical materials of twenty patients with newly treated and refractory recurrent Ph⁺ALL were analyzed retrospectively. After induction treatment with tyrosine kinase inhibitor+vincristine+prednisone（TKI+VP） or tyrosine kinase inhibitor+vincristine+daunorubicin+prednisone（TKI+VDP），17 patients achieved hematological complete remission（HCR） and 3 patients did no remission （NR）. Among the HCR patients， 8 patients for minimal residual disease （MRD） were negative， 3 patients chose Auto-HSCT， and all received TKI maintenance treatment till now after hematopoietic function reconstruction； Allogeneic hematopoietic stem cell transplantation （Allo-HSCT） was performed in other 5 MRD negative patients， 9 MRD positive patients and 3 NR patients.The BCR/ABL fusion genes in the peripheral blood of the patients were detected before transplantation，1 month and 2，3，6，9，and 12 months after stem cell transplantation，and the MRD of the patients was evaluated.After hematopoietic reconstruction in Allo-HSCT group， TKI oral intervention was given regardless of the results of BCR/ABL gene quantitative detection. TKI oral intervention was discontinued after one year for those who continued to test negative. If positive MRD conversion was monitored during the period， effective TKI drugs or immunosuppressant reduction， and other therapeutic measures were given according to the time of transplantation.

Results

All patients achieved hematopoietic reconstruction after transplantation， the median survival time of granulocyte transplantation was 14 （11－24） d， and the median survival time of platelet transplantation was 17 （13－52） d. Follow-up was conducted until December 2020， with a median follow-up time of 52 （3－111） months.The 3-year overall survival （OS） rates of all the patient was （61.1±11.7）%. The 3-year OS rates in Allo-HSCT group and Auto-HSCT group were （52.8±13.4）% and 100.0% （P=0.178），respectively， and the difference was not statistically significant（P=0.178）.The 3-year OS rates of the patients in MRD negative group before treatment and MRD positive group with HCR before treatment were （87.5±11.7）% and （42.8± 15.6）% （P=0.065）， respectively，and the difference was not statistically significant（P=0.065）.All patients with NR before transplantation died.Five cases in whole group died of disease recurrence，1 case of lung infection and 1 case of grade Ⅳ acute graft-versus-host disease.

Conclusion

In the patients with Ph⁺ALL， choosing autologous or allogeneic hematopoietic stem cell transplantation and postoperative stratified intervention according to the dynamic changes of BCR/ABL fusion gene results can improve the OS rate of patients.

Key words: Philadelphia chromosome positive acute lymphoblastic leukemia, Hematopoietic stem cell transplantation, Real-time fluorescence quantitative PCR, fusion gene

中图分类号:

R733.7

冯术青,姚艳红,史月,刘志彬,高峰. 实时荧光定量PCR法检测外周血中BCR/ABL基因表达在Ph⁺急性淋巴细胞白血病患者造血干细胞移植治疗中的应用[J]. 吉林大学学报(医学版), 2022, 48(1): 180-187.

Shuqing FENG,Yanhong YAO,Yue SHI,Zhibin LIU,Feng GAO. Application of real-time fluorescence quantitative PCR for detection of BCR/ABL gene expression in peripheral blood in hematopoietic stem cell transplantation therapy in patients with Ph⁺ acute lymphoblastic leukemia[J]. Journal of Jilin University(Medicine Edition), 2022, 48(1): 180-187.

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