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为了研究急性白血病患者血清及细胞株培养上清中碱性成纤维细胞生长因子 (bFGF)、血管内皮细胞生长因子 (VEGF)的表达水平 ,并初步探讨白血病患者VEGF水平的评价方法以及VEGF特异性反义寡脱氧核苷酸(ASODN)的抗血管生成作用 ,用夹心ELISA法测定患者血清、细胞株 (HL 6 0、U937、NB4、JM和K5 6 2 )培养上清中bFGF和VEGF浓度 ,比较其差异 ,将VEGF血清浓度测定值 (pg ml)、血清浓度测定值经外周血血小板计数标化后标化值VEGF PLT(pg 10 6 )分别作为比较指示 ;HL 6 0细胞经不同浓度VEGFASODN作用后 ,利用噻唑蓝、ELISA法分别测定靶细胞的生长状态及其VEGF分泌水平。结果发现 :①急性白血病患者血清bFGF阳性率 (37.5 % )高于健康对照者 (10 % ) (P <0 .0 1) ,在 5株白血病细胞株中 ,NB4 、U937和K5 6 2细胞上清中检测到了bFGF表达 ;②急性白血病患者及健康者血清VEGF测定值差异无统计学差异 ,但二者标化值测相差显著 (P <0 .0 5 ) ;除U937外 ,其余 4株细胞株培养上清中均有VEGF表达 ;③HL 6 0细胞经 0 .5 ,1及 5 μmol LVEGFASODN作用 2 4小时后 ,其存活率与对照组相比明显降低 (P <0 .0 5 ) ;经 1,5 ,2 0 μmol LVEGFASODN作用 2 4小时后 ,HL 6 0细胞培养上清中VEGF浓度均明显低于对照 (P <0 .0 5 )。结论 :
In order to study the expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in serum and cell culture supernatant of patients with acute leukemia and to evaluate the VEGF level and VEGF specificity in patients with leukemia Antisense oligodeoxynucleotides (ASODNs). The serum concentrations of bFGF and VEGF in serum, cell lines (HL60, U937, NB4, JM and K562) VEGF pT10 (pg106) was compared with the normalized values of VEGF serum concentration (pg ml) and serum concentration measured by peripheral blood platelet count. HL60 cells were treated with different concentrations of VEGFASODN After the treatment, the growth status of target cells and the level of VEGF secretion were measured by using thiazolyl blue and ELISA respectively. The results showed that: ① The positive rate of bFGF in serum of patients with acute leukemia (37.5%) was higher than that of healthy controls (10%) (P <0.01). In 5 leukemia cell lines, NB4, U937 and K562 BFGF expression was detected in Qing; ② There was no significant difference in serum VEGF between acute leukemia patients and healthy subjects, but there was a significant difference between the two values (P <0.05); in addition to U937, the other four cells The survival rate of HL-60 cells treated with 0.5, 1 and 5 μmol LVEGFASODN for 24 hours was significantly lower than that of the control group (P <0.05) The concentration of VEGF in the supernatant of HL-60 cells was significantly lower than that of the control (P <0.05) after treated with 1,5, 20 μmol LVEGFASODN for 24 hours. in conclusion :