Fish检测多发性骨髓瘤染色体异常及预后分析

发布时间：2018-06-06 17:01

本文选题：多发性骨髓瘤 + 常规细胞遗传学　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:应用荧光原位杂交(flurorescence in situ hybridization,FISH)技术检测多发性骨髓瘤(multiple myeloma,MM)患者染色体异常的发生情况及其与MM患者性别、年龄、分期、血常规、生化等临床资料、疗效及预后的关系,明确疾病预后危险分层,探讨其临床意义。方法:收集河北医科大学第三医院血液科自2013年2月1日至2017年1月1日初诊的66例MM患者骨髓标本,进行CD138磁珠分选富集浆细胞后,利用FISH技术,采用特异性探针(包括1q21、TP53、D13S319、IGH/CCND1、IGH/FGFR3、IGH/MAF探针)检测66例MM患者染色体异常的发生情况。结果:1 FISH结果1.1染色体异常的检出率:在66例MM患者中55例出现染色体异常,总异常检出率为83.3%(55/66)。累及探针个数≥2个占56.1%(37/66),用TP53探针检测17号染色体缺失,阳性率为12.1%(8/66);IGH探针检测14号染色体异常,阳性率为54.5%(36/66):其中IGH/MAF阳性率为3.0%(2/66),IGH/FGFR3阳性率为13.6%(9/66),IGH/CCND1阳性率为25.8%(17/66);用D13S319探针检测13号染色体缺失,D13S319阳性率为40.9%(27/66);用1q21探针检测1号染色体异常,1q21扩增阳性率为47.0%(31/66)。1.2 D13S319与IGH之间的关系:D13S319与IGH/FGFR3卡方检验发现D13S319阳性患者常伴随IGH/FGFR3阳性,具有统计学意义(P=0.0200.05),其他染色体异常之间无相关性。1.3染色体异常与患者临床资料之间的关系:1q21扩增与HGB降低有关(P=0.028);TP53与LDH升高有关(P=0.041),与首次化疗后感染有关(P=0.017);D13S319与高年龄有关(P=0.020);IGH/CCND1与骨损伤数量增加有关(P=0.003),与白蛋白减少有关(P=0.049);IGH/FGFR3与白蛋白减少有关(P=0.010)。其他染色体异常与MM患者临床资料无关。2 FISH结果与患者生存期关系随访至2017年1月1日13例(19.7%)患者死亡。FISH结果采用Kaplan-Meier法计算生存率并绘制生存曲线,单因素分析发现TP53阳性(8例)和阴性(53例),OS分别为28.250和42.605个月,PFS分别为17.857和28.415个月,具有统计学意义(P=0.027和P=0.045);1q21异常者(31例)PFS明显短于正常者(30例),PFS为21.017和31.896个月,具有统计学意义(P=0.022),但OS无统计学差异(P=0.622);IGH/FGFR3异常者(9例)和正常者(50例),OS分别为17.889和41.672个月,PFS分别为12.063和28.757个月,具有统计学意义(P=0.037和P=0.025);IGH/MAF基因异常者(2例)和正常者(57例),OS分别为7.500和41.457个月,不具有统计学差异(P=0.137),PFS分别为4.000和28.518个月,具有统计学差异(P=0.000);D13S319基因异常者(25例)和正常者(36例)OS分别为39.926和37.926个月,PFS分别为28.466和19.270个月,差异无统计学意义(P=0.700和P=0.182)。将1q21探针检测的61例患者分为三组:1q21阴性组、单纯1q21阳性、1q21阳性并伴有其他染色体异常者,OS分别为43.618、24.314和13.500个月,无统计学差异(P0.05);PFS分别为31.896、20.400和8.125个月,1q21阴性组和单纯1q21阳性之间存在统计学差异(P=0.038),其他无统计学差异(P0.05)。3临床资料与患者生存期关系3.1 ISS、R-ISS分期与预后的关系:根据ISS分期,Ⅰ期5例,Ⅱ期23例,Ⅲ期38例,OS分别为41.000、34.436和35.775个月,Ⅰ期Ⅱ期Ⅲ期,PFS分别为23.250、27.647和25.768个月,差异均无统计学意义(P=0.918和P=0.918);根据R-ISS分期,Ⅰ期5例,Ⅱ期47例,Ⅲ期14例,OS分别为47.022、37.585和20.674个月;PFS分别为23.250、30.005和13.464个月,Ⅲ期PFS较Ⅰ期/Ⅱ期明显缩短,差异均具有统计学意义(P=0.039和P=0.000);根据m SMART分期,OS低危41.771个月中危38.599个月高危22.111个月,PFS低危31.055个月中危20.045个月高危16.375个月,高危PFS较低危明显缩短,差异均有统计学意义(P=0.036和P=0.010)。3.2临床指标和预后的关系:HB100g/L较≥100g/L者OS为31.862和44.324个月,无统计学差异(P=0.136);PFS分别为20.935和30.552个月,具有统计学差异(P=0.020)。PLT100×109/L较≥100×109/L者OS明显缩短,OS分别为11.906和43.738个月,具有统计学差异(P=0.000);PFS分别为15.641和28.029个月,无统计学差异(P=0.069)。LDH≥225U/L较225U/L者OS、PFS缩短,OS分别为23.375和44.601、PFS分别为15.631和29.217个月,具有统计学差异(P=0.003和P=0.002)。首次化疗具有感染的患者OS较未感染的患者明显缩短,分别为30.236和47.917个月,具有统计学意义(P=0.005);PFS无统计学差异(P=0.673)。3.3预后多因素分析发现m SMART高危患者、浆细胞≥30%、PLT100×109/L、ALB35g/L、LDH≥225U/L可以使OS缩短;R-ISSⅢ期、HB100g/L、PLT100×109/L、Cr≥175umol/L、首次化疗存在感染的患者PFS缩短。结论:1 FISH方法发现MM患者最常见的基因异常为IGH易位,染色体异常多发生在在14、1、13号染色体。2 D13S319阳性患者常伴随IGH/FGFR3阳性。3 1q21扩增与HGB降低有关;TP53与LDH升高有关,与首次化疗后感染有关;D13S319与高年龄有关;IGH/CCND1与骨损伤数量增加有关,与白蛋白减少有关;IGH/FGFR3与白蛋白减少有关。其他染色体异常与MM患者临床资料无关。4 TP53、IGH/FGFR3和IGH/MAF是不良的预后因素,OS和PFS均具有统计学意义;D13S319 OS和PFS差异均无统计学意义;1q21阳性和阴性组PFS具有统计学意义,但OS无统计学差异;1q21阴性组和单纯1q21阳性之间PFS存在统计学差异。5 ISS分期OS和PFS差异均无统计学意义;R-ISS分期OS和PFS差异均具有统计学意义;m SMART分期,高危OS和PFS较低危明显缩短,差异均有统计学意义。6 HB100g/L较≥100g/L者OS无统计学差异,PFS具有统计学差异;PLT100×109/L较≥100×109/L者OS明显缩短,OS具有统计学差异,PFS无统计学差异;LDH≥225U/L较225U/L者OS、PFS缩短,均具有统计学差异;首次化疗具有感染的患者OS较未感染的患者明显缩短,具有统计学意义,PFS无统计学差异。7预后多因素分析发现m SMART高危患者、浆细胞≥30%、PLT100×109/L、ALB35g/L、LDH≥225U/L可以使OS缩短;R-ISSⅢ期、HB100g/L、PLT100×109/L、Cr≥175umol/L、首次化疗存在感染的患者PFS缩短。
[Abstract]:Objective: to detect the occurrence of chromosomal abnormalities in patients with multiple myeloma (multiple myeloma, MM) by Flurorescence in situ hybridization (FISH) and their relationship with the sex, age, staging, blood routine, biochemical and other clinical data, the effect and prognosis of the patients with multiple myeloma, and to clarify the risk stratification of the prognosis of the disease. Methods: the bone marrow specimens of 66 MM patients from February 1, 2013 to January 1, 2017 in the Department of Hematology, Third Hospital of Hebei Medical University, were collected to enrich the plasma cells by CD138 magnetic beads sorting. The specific probes (including 1q21, TP53, D13S319, IGH/CCND1, IGH/FGFR3, IGH/MAF probe) were used to detect the staining of 66 cases of MM patients. Results: 1 FISH results: the detection rate of 1.1 chromosome abnormality: in 66 cases of MM, 55 cases had chromosomal abnormalities, the total abnormal detection rate was 83.3% (55/66). The number of probe number more than 2 accounted for 56.1% (37/66), and the TP53 probe was used to detect the deletion of chromosome 17, the positive rate was 12.1% (8/66), and the IGH probe detected chromosome 14 abnormality, Yang The rate of sex was 54.5% (36/66), of which the positive rate of IGH/MAF was 3% (2/66), the positive rate of IGH/FGFR3 was 13.6% (9/66), and the positive rate of IGH/CCND1 was 25.8% (17/66); the chromosome deletion of chromosome 13 was detected with D13S319 probe and the positive rate of D13S319 was 40.9% (27/66). The 1q21 probe was used to detect chromosome 1, and the positive rate of 1q21 amplification was 47%. Relationship: D13S319 and IGH/FGFR3 chi square test found that D13S319 positive patients were often accompanied by IGH/FGFR3 positive, statistically significant (P=0.0200.05), other chromosomal abnormalities associated with the relationship between abnormal.1.3 chromosomes and the patient's clinical data: 1q21 amplification was associated with HGB reduction (P=0.028); TP53 was associated with LDH elevation (P=0.041), and first Infection related after secondary chemotherapy (P=0.017); D13S319 was associated with high age (P=0.020); IGH/CCND1 was associated with increased number of bone injuries (P=0.003), associated with albumin reduction (P=0.049); IGH/FGFR3 was associated with albumin reduction (P=0.010). Other chromosomal abnormalities were not related to clinical data in patients with MM; the relationship between.2 FISH and patient survival was followed up to 1 in 2017. On the 1 day, 13 cases (19.7%) patients died.FISH results using Kaplan-Meier method to calculate survival rate and draw survival curve. Single factor analysis found that TP53 positive (8 cases) and negative (53 cases), OS were 28.250 and 42.605 months respectively, PFS was 17.857 and 28.415 months respectively, with statistical significance (P=0.027 and P=0.045); 1q21 abnormal (31 cases) PFS obviously shorter than The normal (30 cases), PFS for 21.017 and 31.896 months, had statistical significance (P=0.022), but there was no statistical difference in OS (P=0.622); abnormal IGH/FGFR3 (9 cases) and normal (50 cases), OS were 17.889 and 41.672 months respectively, PFS was 12.063 and 28.757 months respectively, P=0.037 and P=0.025 (2 cases) and normal (2 cases) and normal. (57 cases), OS was 7.500 and 41.457 months respectively, with no statistical difference (P=0.137), PFS was 4 and 28.518 months, respectively, with statistical difference (P=0.000); D13S319 gene abnormal (25 cases) and normal (36 cases) OS were 39.926 and 37.926 months respectively, PFS was 28.466 and 19.270 months respectively, the difference was not statistically significant (P=0.700 and P=0.182). 61 patients with 1q21 probe were divided into three groups: 1q21 negative group, simple 1q21 positive, 1q21 positive and other chromosomal abnormality, OS was 43.618,24.314 and 13.500 months respectively, without statistical difference (P0.05); PFS was 31.896,20.400 and 8.125 months respectively, 1q21 negative group and simple 1q21 positive were statistically different (P=0.038), Other non statistical difference (P0.05).3 clinical data and patient survival relationship 3.1 ISS, the relationship between R-ISS staging and prognosis: according to ISS stage, stage I 5 cases, 23 cases in stage II, 38 cases in stage III, OS respectively 41.000,34.436 and 35.775 months, stage I stage III, PFS respectively 23.250,27.647 and 25.768 months, the difference is not statistically significant (P=0.918 and P=0. .918); according to R-ISS staging, 5 cases, 47 cases in stage II, 14 cases in stage III, OS for 47.022,37.585 and 20.674 months respectively; PFS was 23.250,30.005 and 13.464 months respectively. The stage III PFS was significantly shorter than phase I / II, the difference was statistically significant (P=0.039 and P=0.000), the root m SMART staging, and the low risk 41.771 months of OS low risk 38.599 months risk 22.111 The risk of high risk for 20.045 months was 16.375 months in the low risk of PFS 31.055 months, and the high risk of PFS was significantly shorter than that of low risk. The difference was statistically significant (P=0.036 and P=0.010).3.2 clinical indicators and prognosis: HB100g/L was more than 100g/L in OS for 31.862 and 44.324 months, with no statistical difference (P=0.136); PFS was 20.935 and 30.552 months, respectively. The study difference (P=0.020).PLT100 x 109/L was significantly shorter than that of 100 x 109/L, OS was 11.906 and 43.738 months respectively, with statistical difference (P=0.000); PFS was 15.641 and 28.029 months respectively. There was no statistical difference (P=0.069).LDH > 225U/L compared with 225U/L, 23.375 and 44.601 respectively, respectively, 15.631 and 29.217 months respectively. Statistical differences (P=0.003 and P=0.002). OS was significantly shorter in patients with infection than in uninfected patients, 30.236 and 47.917 months, respectively, with statistical significance (P=0.005). There was no statistical difference in PFS (P=0.673).3.3 prognosis multivariate analysis found that m SMART high risk patients, plasma cells more than 30%, PLT100 x 109/L, ALB35g/L, or more than It is possible to shorten OS, R-ISS III, HB100g/L, PLT100 * 109/L, Cr > 175umol/L, and PFS shortening in patients with infection in the first chemotherapy. Conclusion: the 1 FISH method found that the most common gene abnormality in MM patients is IGH translocation. Low related; TP53 is associated with elevated LDH, associated with first chemotherapy after chemotherapy; D13S319 is associated with high age; IGH/CCND1 is associated with increased number of bone injuries, associated with albumin reduction; IGH/FGFR3 is associated with albumin reduction. Other chromosomal abnormalities are not related to the clinical data of MM patients.4 TP53, IGH/FGFR3 and IGH/MAF are adverse prognostic factors, OS and PFS. There was no statistical significance in both D13S319 OS and PFS, while PFS in 1q21 positive and negative group had statistical significance, but there was no statistical difference between OS and 1q21 negative group and 1q21 positive. There was no statistical difference between.5 ISS and 1q21 positive..5 ISS stage OS and differences were not statistically significant. M SMART staging, high risk OS and PFS were significantly shorter than low risk. There was no statistical difference between.6 HB100g/L and 100g/L in OS, PFS had statistical difference, and PLT100 x 109/L was shorter than that of 100 x 109/L. Differences; OS was significantly shorter than those in uninfected patients for the first time chemotherapy. There was statistical significance. There was no statistical difference in PFS. The multivariate analysis of prognostic factors of.7 found that m SMART was more than 30%, PLT100 x 109/L, ALB35g/L, LDH > 225U/L could shorten the OS; PFS in patients with infection was shortened.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.3

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