急性淋巴细胞白血病患者FGFR3表达对循环内皮细胞增殖的影响

摘要

目的: 了解急性淋巴细胞白血病(ALL)患者成纤维细胞生长因子受体3(FGFR3)表达对循环内皮细胞(CECs)增殖的影响.方法: 通过RT-PCR法检测44例ALL患者骨髓中FGFR3 mRNA表达,分成FGFR3+组及FGFR3-组进行Kaplan-Meier生存分析.利用免疫磁珠结合流式细胞术分离计数CECs,对比两组CECs数量、表面抗原表达、生长曲线及集落生成数的差异.免疫荧光组化染色测定CECs表面FGFR3表达水平.结果: 44例核型正常ALL患者中FGFR3 mRNA表达阳性率为43.2%,T-ALL组的FGFR3表达高于B-ALL组(P<0.05).19 d骨髓原始细胞比例≥5%的患者FGFR3表达升高(P<0.05).FGFR3阳性组的总生存率明显低于阴性组(P<0.05).分离出的CECs高表达CD31、CD144、VEGFR-2和CD146,基本不表达CD45.与FGFR3阴性组相比,阳性组CECs数量、CD133的阳性率及集落生成数均增多(P<0.05).体外培养中,FGFR3阳性组与阴性组相比,生长曲线3个时点上的细胞数差异有统计学显著性(P<0.05).19例ALL-FGFR3+患者CECs表面均能检测到FGFR3表达,阳性率为29.00%±15.71%.结论: 致癌基因FGFR3对ALL患者CECs的增殖有促进作用,可能具备了抗肿瘤及抗血管生成的双重靶点身份,可以为今后的分子治疗提供更多的选择.%AIM: To evaluate the expression of fibroblast growth factor receptor 3 (FGFR3) in acute lymphocytic leukemia (ALL) patients and its contribution to the proliferation of circulating endothelial cells (CECs).METHODS: The mRNA expression levels of FGFR3 in 44 patients with ALL were assayed by RT-PCR.Overall survival (OS) rates of the patients in FGFR3+ group and FGFR3-group were estimated by Kaplan-Meier analysis.The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cell counts, antigen expression, growth curve and colony forming rate of the CECs in the 2 groups were determined.The FGFR3 expression of CECs was identified by immunofluorescence staining.RESULTS: The positive rate of FGFR3 mRNA expression was 43.2% in 44 ALL patients with normal karyotype.T-ALL expressed higher level of FGFR3 than B-ALL (P<0.05).FGFR3 was over-expressed in ALL patients with bone marrow blast proportion ≥5% (P<0.05).The probability of OS was significantly lower in FGFR3+ group than that in FGFR3-group (P<0.05).The sorted CECs highly expressed CD31, CD144, VEGFR-2 and CD146, and rarely expressed CD45.The counts of CECs and expression level of CD133 significantly increased in FGFR3+ group compared with FGFR3-group.The same result of the amount of colony formation was observed (P<0.05).There was significant difference at 3 time points of cultured CECs count in vitro between FGFR3+ group and FGFR3-group (P<0.05).The positive rate of FGFR3 expression of CECs from 19 ALL-FGFR3+ patients was (29.00±15.71)%.CONCLUSION: The over-expression of FGFR3 gene in ALL may be helpful to evaluate the prolife-ration of CECs, and become a double target with anti-tumor and anti-angiogenesis effects to offer more choice for molecular therapy in the future.