急性白血病患者血管内皮生长因子和碱性成纤维细胞生长因子的测定及临床意义
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摘要
目的: 血管内皮生长因子（Vascular endothelial growth factor VEGF）和碱性成纤维细胞生长因子 (basic fibroblast growth factor, bFGF)是两种重要的血管源性生长因子，不仅具有重要的生理作用，而且与实体瘤的生长、扩散、转移密切相关。VEGF可强烈的促进血管内皮细胞（VEC）的增殖、分裂和迁移；增加微血管的通透性，使肿瘤血管通透性增高；通过旁分泌方式作用于邻近的VEC。bFGF在体内分布广泛，能促进表皮内皮细胞的再生，促进血管内皮细胞分裂，刺激血管内皮细胞向肿瘤组织趋化运动并形成管状结构，还能抑制活化的自然杀伤细胞对肿瘤血管内皮细胞的粘附，这表明bFGF可以为肿瘤血管提供保护。近年来VEGF、bFGF在造血系统恶性疾病中的作用日益受到重视，国内外的大量研究表明VEGF、bFGF与白血病的发病机制、临床特征及预后密切相关。本文采用敏感的酶联免疫吸附法（emzyme linked immuno-sorbent assay, ELISA）检测急性髓性白血病（AML）、急性淋巴细胞白血病（ALL）患者和健康人群中的血清VEGF和bFGF的含量及AML患者和正常对照骨髓培养上清液中的VEGF含量，同时检测临床相关化验指标，并对所测数据进行统计分析，目的在于研究VEGF和bFGF在急性白血病中是否呈现过渡表达，探讨将其作为白血病
转归及预后指标的可行性，试图为临床的诊断、治疗及疾病的预后判断提供简便可靠的观察指标。
方法：选择初治的30例急性髓性白血病患者，26例急性淋巴细胞白血病患者，16例血清对照组（来自职工健康查体），20例骨髓对照组（骨髓形态学及病理学检查正常的白细胞减少症和未累及骨髓的实体肿瘤患者），用酶联免疫吸附试验（ELISA）检测AML、ALL患者及正常对照组血清VEGF、bFGF水平，检测AML患者及正常对照骨髓培养上清液中VEGF水平，并比较VEGF、bFGF在治疗前后的变化及与临床化验指标之间的相关性。所有数据均采用SPSS10.0统计软件分析。
结果：1 AML、ALL患者及正常对照组血清VEGF及bFGF水平的比较：(1) AML患者的血清VEGF（s-VEGF）浓度为(275.22±229.69)pg/ml,显著高于正常对照组(121.13±104.62)pg/ml(p<0.01)；血清bFGF(s-bFGF)浓度为(15.89±14.20)pg/ml,显著高于正常对照组(8.15±7.66)pg/ml(p<0.05)。(2) ALL患者的s-VEGF浓度为(248.73 ±245.17)pg/ml, 显著高于正常对照组(121.13±104.62)pg/ml(p<0.01), s-bFGF浓度为(15.48±12.83)pg/ml, 显著高于正常对照组(8.15±7.66)pg/ml(p<0.05)。2 AML、ALL患者化疗前后 VEGF及bFGF水平的比较：（1）将AML患者根据第一次化疗的疗效分为完全缓解组(CR组)和未完全缓解组（NR组），结果显示CR组和NR组治疗前s-VEGF量为（248.73±245.17）pg/ml和（485.08±271.38）pg/ml，NR组明显高于CR组，两组比较有统计学意义（P<0.05）； CR组和NR组治疗前s-bFGF量为
（15.64±13.54）pg/ml和（16.71±17.36）pg/ml，两组比较NR组高于CR组，但是无统计学意义（P>0.05）。(2)同样将ALL患者分为CR组和NR组，结果显示CR组和NR组治疗前s-VEGF量为（184.09±193.55）pg/ml和（520.23±272.59）pg/ml，NR组明显高于CR组,有统计学意义（P<0.01）； CR组和NR组治疗前s-bFGF量为（12.01±10.13）pg/ml和（30.41±13.68）pg/ml， NR组高于CR组，有统计学意义（P<0.01）。3 AML患者和对照组骨髓培养上清VEGF的水平的比较、治疗前后的变化及与临床化验指标和生存期的关系：（1）骨髓单个核细胞培养72小时后，VEGF的分泌量为(425.31±167.27)pg/ml，对照组则为（140.12±75.13）pg/ml， AML患者显著高于对照组，有统计学意义（P<0.001）。（2）我们将患者根据第一次化疗的疗效分为CR组和NR组，结果显示CR组和NR组治疗前VEGF量为（387.93±150.37）pg/ml和（562.39±165.59）pg/ml，两组比较有统计学意义（P<0.05）。（3）化疗前患者血清乳酸脱氢酶（LDH）、血清β2微球蛋白（β2-MG）、骨髓原始细胞数、血红蛋白（Hb）、血小板(PLT)与骨髓培养上清VEGF水平均无相关性（p>0.05， Pearson 相关检验）。(4)为研究VEGF分泌量与患者生存期的关系，将VEGF以其均值（425.31pg/ml）为界分为VEGF较低组（小于均值）和较高组（大于均值）。Kaplan-Meier分析显示，治疗前VEGF较低组的生存时间大于VEGF较高组（分别为16.9月和6.2个月，P<0.001）。4 AML、ALL患者s-VEGF、bFGF水平之间的相关性及与临床化验指标的相关性。（1）AML患者s-VEGF水平
与骨髓单个核细胞培养上清VEGF水平及s-bFGF水平均成正相关（P<0.05， Pearson 相关检验）。ALL患者s-VEGF水平与s-bFGF水平成正相关，（P<0.01， Pearson 相关检验）。（2） AML、ALL患者s-VEGF、bFGF水平与LDH、血β2MG、血小板、骨髓原始细胞、Hb均无相关性（P>0.05， Pearson 相关检验）。5. AML、ALL患者s-VEGF、bFGF的水平与生存期的关系：（1）为研究AML 患者s-VEGF、bFGF水平与患者生存期的关系，将患者s-VEGF水平以其均值（248.73pg/ml）为界分为VEGF较低组（小于均值）和较高组（大于均值）。Kaplan-Meier分析显示，治疗前VEGF较低组的生存时间大于VEGF较高组（分别为12.83个月和6.20个月，P<0.01）；
Objective: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)are two essential angiogenic growth factors, not only having important physiologic function, but also playing critical roles in solid tumor development and metastasis. VEGF can promote proliferation, split and migration of vascular endothelial cells (VEC); improve permeability of microvessel and increase permeability of tumor vessel to plasma proteins; have function on near VEC by paracrine. BFGF is distributed over living body and can give tumor cell protection because it can promote epidermis endothelial cell to reproduce and promote to split, can stimulate vascular endothelial cells converting tumor cell and becoming tube structure, can inhibit NK cell to adhere to tumor cell. In recent year the function of VEGF and bFGF in hemic malignancies become more and more important and lots of studies showed VEGF, bFGF have closed relationship with leukemia. We detected concentration of serum VEGF and bFGF in acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and healthy control and concentration of VEGF of bone marrow
monocyte cell culture supernatant in AML and healthy control by emzyme linked immuno-sorbent assay ( ELISA). In the same time we detected some clinical chemical examination and analyzed them with statistic. Our objectives is to study if VEGF and bFGF have over expression in leukemia , discuss the possibility of VEGF and bFGF as predictor of outcome in leukemia and try to find simple, depended indices for clinical diagnose and therapy.
Methods: Pretreatment serum VEGF and bFGF concentration were measured in peripheral blood serum obtained from 30 patients with newly diagnosed AML, 26 patients with newly diagnosed ALL and 16 healthy controls ; VEGF concentration of marrow monocyte cell culture supernatant in AML and healthy control also was measured in 28 AML patients and 20 healthy controls. All data were analyzed by SPSS10.0. P Values less than 0.05 were considered to be statistically significant.
Results: 1.The comparison of serum VEGF and bFGF concentration in AML , ALL patients and healthy control (1)Serum VEGF(s-VEGF) levels were elevated in AML patients(275.22±229.69pg/ml) compared to control healthy(121.13±104.62pg/ml) (p<0.01); Serum bFGF(s-bFGF) levels were elevated in AML patients(15.89±14.20pg/ml), compared to control healthy(8.15±7.66pg/ml) (p<0.05).(2)S-VEGF levels were (248.73 ±245.17)pg/ml in ALL patients, significantly higher than
healthy controls (121.13±104.62pg/ml) (p<0.01); S-bFGF levels were (15.48±12.83)pg/ml in ALL patients, significantly higher than healthy controls (8.15±7.66pg/ml) (p<0.05)。2.The comparison between s-VEGF and bFGF concentration in AML , ALL patients pre- and post- treatment.(1)We divided AML patients into two groups(completely release CR and no completely release NR) depended on the reaction on first chemical therapy and found s-VEGF levels（248.73±245.17pg/ml ）of pretreatment patients in CR group were lower significantly than in NR(485.08±271.38pg/ml)（P<0.05）; s-bFGF levels（15.64±13.54pg/ml）of pretreatment patients in CR group were lower significantly than in NR（16.71±17.36）pg/ml), But it is not statistically significant（P>0.05）.(2) We found s-VEGF levels（184.09±193.55pg/ml）of pretreatment patients in CR group were lower significantly than in NR(520.23±272.59pg/ml)（P<0.05）; s-bFGF levels（12.01±10.13pg/ml）of pretreatment patients in CR group were lower significantly than in NR（30.41±13.68pg/ml） （P<0.01）.3. The comparison between VEGF concentration of marrow monocyte cell culture supernatant in AML and healthy controls ; The comparison between pre -and post -treatment in AML patients; The relationship between VEGF concentration and other chemical examinations: (1) VEGF levels in culture supernatant of marrow monocyte cell cultured 72 hours were (425.31±167.27)pg/ml, significantly
higher than healthy controls( 140.12±75.13pg/ml) （P<0.001）.(2) We divided AML patients into t
转归及预后指标的可行性，试图为临床的诊断、治疗及疾病的预后判断提供简便可靠的观察指标。
方法：选择初治的30例急性髓性白血病患者，26例急性淋巴细胞白血病患者，16例血清对照组（来自职工健康查体），20例骨髓对照组（骨髓形态学及病理学检查正常的白细胞减少症和未累及骨髓的实体肿瘤患者），用酶联免疫吸附试验（ELISA）检测AML、ALL患者及正常对照组血清VEGF、bFGF水平，检测AML患者及正常对照骨髓培养上清液中VEGF水平，并比较VEGF、bFGF在治疗前后的变化及与临床化验指标之间的相关性。所有数据均采用SPSS10.0统计软件分析。
结果：1 AML、ALL患者及正常对照组血清VEGF及bFGF水平的比较：(1) AML患者的血清VEGF（s-VEGF）浓度为(275.22±229.69)pg/ml,显著高于正常对照组(121.13±104.62)pg/ml(p<0.01)；血清bFGF(s-bFGF)浓度为(15.89±14.20)pg/ml,显著高于正常对照组(8.15±7.66)pg/ml(p<0.05)。(2) ALL患者的s-VEGF浓度为(248.73 ±245.17)pg/ml, 显著高于正常对照组(121.13±104.62)pg/ml(p<0.01), s-bFGF浓度为(15.48±12.83)pg/ml, 显著高于正常对照组(8.15±7.66)pg/ml(p<0.05)。2 AML、ALL患者化疗前后 VEGF及bFGF水平的比较：（1）将AML患者根据第一次化疗的疗效分为完全缓解组(CR组)和未完全缓解组（NR组），结果显示CR组和NR组治疗前s-VEGF量为（248.73±245.17）pg/ml和（485.08±271.38）pg/ml，NR组明显高于CR组，两组比较有统计学意义（P<0.05）； CR组和NR组治疗前s-bFGF量为
（15.64±13.54）pg/ml和（16.71±17.36）pg/ml，两组比较NR组高于CR组，但是无统计学意义（P>0.05）。(2)同样将ALL患者分为CR组和NR组，结果显示CR组和NR组治疗前s-VEGF量为（184.09±193.55）pg/ml和（520.23±272.59）pg/ml，NR组明显高于CR组,有统计学意义（P<0.01）； CR组和NR组治疗前s-bFGF量为（12.01±10.13）pg/ml和（30.41±13.68）pg/ml， NR组高于CR组，有统计学意义（P<0.01）。3 AML患者和对照组骨髓培养上清VEGF的水平的比较、治疗前后的变化及与临床化验指标和生存期的关系：（1）骨髓单个核细胞培养72小时后，VEGF的分泌量为(425.31±167.27)pg/ml，对照组则为（140.12±75.13）pg/ml， AML患者显著高于对照组，有统计学意义（P<0.001）。（2）我们将患者根据第一次化疗的疗效分为CR组和NR组，结果显示CR组和NR组治疗前VEGF量为（387.93±150.37）pg/ml和（562.39±165.59）pg/ml，两组比较有统计学意义（P<0.05）。（3）化疗前患者血清乳酸脱氢酶（LDH）、血清β2微球蛋白（β2-MG）、骨髓原始细胞数、血红蛋白（Hb）、血小板(PLT)与骨髓培养上清VEGF水平均无相关性（p>0.05， Pearson 相关检验）。(4)为研究VEGF分泌量与患者生存期的关系，将VEGF以其均值（425.31pg/ml）为界分为VEGF较低组（小于均值）和较高组（大于均值）。Kaplan-Meier分析显示，治疗前VEGF较低组的生存时间大于VEGF较高组（分别为16.9月和6.2个月，P<0.001）。4 AML、ALL患者s-VEGF、bFGF水平之间的相关性及与临床化验指标的相关性。（1）AML患者s-VEGF水平
与骨髓单个核细胞培养上清VEGF水平及s-bFGF水平均成正相关（P<0.05， Pearson 相关检验）。ALL患者s-VEGF水平与s-bFGF水平成正相关，（P<0.01， Pearson 相关检验）。（2） AML、ALL患者s-VEGF、bFGF水平与LDH、血β2MG、血小板、骨髓原始细胞、Hb均无相关性（P>0.05， Pearson 相关检验）。5. AML、ALL患者s-VEGF、bFGF的水平与生存期的关系：（1）为研究AML 患者s-VEGF、bFGF水平与患者生存期的关系，将患者s-VEGF水平以其均值（248.73pg/ml）为界分为VEGF较低组（小于均值）和较高组（大于均值）。Kaplan-Meier分析显示，治疗前VEGF较低组的生存时间大于VEGF较高组（分别为12.83个月和6.20个月，P<0.01）；
Objective: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)are two essential angiogenic growth factors, not only having important physiologic function, but also playing critical roles in solid tumor development and metastasis. VEGF can promote proliferation, split and migration of vascular endothelial cells (VEC); improve permeability of microvessel and increase permeability of tumor vessel to plasma proteins; have function on near VEC by paracrine. BFGF is distributed over living body and can give tumor cell protection because it can promote epidermis endothelial cell to reproduce and promote to split, can stimulate vascular endothelial cells converting tumor cell and becoming tube structure, can inhibit NK cell to adhere to tumor cell. In recent year the function of VEGF and bFGF in hemic malignancies become more and more important and lots of studies showed VEGF, bFGF have closed relationship with leukemia. We detected concentration of serum VEGF and bFGF in acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and healthy control and concentration of VEGF of bone marrow
monocyte cell culture supernatant in AML and healthy control by emzyme linked immuno-sorbent assay ( ELISA). In the same time we detected some clinical chemical examination and analyzed them with statistic. Our objectives is to study if VEGF and bFGF have over expression in leukemia , discuss the possibility of VEGF and bFGF as predictor of outcome in leukemia and try to find simple, depended indices for clinical diagnose and therapy.
Methods: Pretreatment serum VEGF and bFGF concentration were measured in peripheral blood serum obtained from 30 patients with newly diagnosed AML, 26 patients with newly diagnosed ALL and 16 healthy controls ; VEGF concentration of marrow monocyte cell culture supernatant in AML and healthy control also was measured in 28 AML patients and 20 healthy controls. All data were analyzed by SPSS10.0. P Values less than 0.05 were considered to be statistically significant.
Results: 1.The comparison of serum VEGF and bFGF concentration in AML , ALL patients and healthy control (1)Serum VEGF(s-VEGF) levels were elevated in AML patients(275.22±229.69pg/ml) compared to control healthy(121.13±104.62pg/ml) (p<0.01); Serum bFGF(s-bFGF) levels were elevated in AML patients(15.89±14.20pg/ml), compared to control healthy(8.15±7.66pg/ml) (p<0.05).(2)S-VEGF levels were (248.73 ±245.17)pg/ml in ALL patients, significantly higher than
healthy controls (121.13±104.62pg/ml) (p<0.01); S-bFGF levels were (15.48±12.83)pg/ml in ALL patients, significantly higher than healthy controls (8.15±7.66pg/ml) (p<0.05)。2.The comparison between s-VEGF and bFGF concentration in AML , ALL patients pre- and post- treatment.(1)We divided AML patients into two groups(completely release CR and no completely release NR) depended on the reaction on first chemical therapy and found s-VEGF levels（248.73±245.17pg/ml ）of pretreatment patients in CR group were lower significantly than in NR(485.08±271.38pg/ml)（P<0.05）; s-bFGF levels（15.64±13.54pg/ml）of pretreatment patients in CR group were lower significantly than in NR（16.71±17.36）pg/ml), But it is not statistically significant（P>0.05）.(2) We found s-VEGF levels（184.09±193.55pg/ml）of pretreatment patients in CR group were lower significantly than in NR(520.23±272.59pg/ml)（P<0.05）; s-bFGF levels（12.01±10.13pg/ml）of pretreatment patients in CR group were lower significantly than in NR（30.41±13.68pg/ml） （P<0.01）.3. The comparison between VEGF concentration of marrow monocyte cell culture supernatant in AML and healthy controls ; The comparison between pre -and post -treatment in AML patients; The relationship between VEGF concentration and other chemical examinations: (1) VEGF levels in culture supernatant of marrow monocyte cell cultured 72 hours were (425.31±167.27)pg/ml, significantly
higher than healthy controls( 140.12±75.13pg/ml) （P<0.001）.(2) We divided AML patients into t
引文
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