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目的建立人髓性白血病细胞血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)及其受体KDR和Flt-1在基因转录和蛋白质水平上的检测方法,为临床判断白血病患者的预后提供技术支持。方法用RT-PCR测定K562和HL-602种白血病细胞VEGF及其受体mRNA表达,免疫组织化学染色法检测VEGF及其受体蛋白表达;ELISA法检测细胞上清液中VEGF的分泌水平。结果2种髓系白血病细胞均检测到VEGF的基因转录和蛋白表达。HL-60细胞同时表达Flt-1和KDR2种受体,而K562细胞仅有受体Flt-1的表达,未见受体KDR的mRNA和蛋白表达。细胞培养上清液中检出VEGF的高分泌。结论VEGF及其受体在髓性白血病细胞表达增高,但不同细胞系VEGF受体表达有差异。VEGF及其受体在白血病的发生、发展中可能起重要作用。
OBJECTIVE: To establish a method for the detection of vascular endothelial growth factor (VEGF) and its receptors KDR and Flt-1 in human myeloid leukemia cells at gene transcription and protein level, and to provide technical support for the clinical judgment of the prognosis of leukemia patients. Methods The expression of VEGF and its receptor mRNA in K562 and HL-602 leukemia cells was detected by RT-PCR. The expression of VEGF and its receptor protein was detected by immunohistochemical staining. The secretion of VEGF in the supernatant was detected by ELISA. Results Both myeloid leukemia cells detected VEGF gene transcription and protein expression. HL-60 cells express both Flt-1 and KDR2 receptors, while K562 cells only express Flt-1, but no receptor KDR mRNA and protein expression. High secretion of VEGF was detected in cell culture supernatants. Conclusion The expression of VEGF and its receptor in myeloid leukemia cells increased, but the expression of VEGF receptor in different cell lines was different. VEGF and its receptors may play an important role in the occurrence and development of leukemia.