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血管内皮细胞生长因子 (VEGF)是血管内皮细胞特异性的丝裂原 ,与实体肿瘤的血管生成、生长、转移及不良预后密切相关。最近的报道表明 ,慢性髓性白血病 (CML)病人骨髓过度表达VEGF。然而 ,VEGF在CML细胞异常增殖中的作用还不清楚。为了探讨自分泌VEGF对CML细胞系K5 62增殖与凋亡的影响 ,本研究首次采用正义和反义VEGF12 1cDNA转染K5 62细胞 ,通过RT PCR及ELISA方法鉴定各转染克隆VEGFmRNA及蛋白的表达。结果显示 ,转染反义VEGF12 1cDNA可以降低K5 62细胞VEGF的表达 ,而转染正义VEGF12 1cDNA可使VEGF的表达提高 3倍以上。MTT ,集落形成试验及细胞凋亡检测试验观察 :反义VEGF12 1cDNA转染可以抑制K5 62细胞的增殖及其集落形成 ,而促进其凋亡 ;正义VEGF12 1cDNA转染与其结果相反。本研究结果说明自分泌VEGF在CML细胞系K5 62的增殖与凋亡中起重要作用
Vascular endothelial growth factor (VEGF) is a mitogen specific to vascular endothelial cells and is closely related to angiogenesis, growth, metastasis and poor prognosis of solid tumors. Recent reports indicate that bone marrow is overexpressing VEGF in patients with chronic myelogenous leukemia (CML). However, the role of VEGF in the abnormal proliferation of CML cells remains unclear. To investigate the effect of autocrine VEGF on the proliferation and apoptosis of CML cell line K5 62, K5 62 cells were transfected with sense and antisense VEGF 12 cDNA for the first time. The expression of VEGF mRNA and protein in each transfected clone was identified by RT PCR and ELISA. . The results showed that transfection of antisense VEGF12 1 cDNA can reduce the expression of VEGF in K5 62 cells, and transfection of sense VEGF12 1 cDNA can increase the expression of VEGF by more than 3 times. MTT, colony formation assay and apoptosis detection test: antisense VEGF12 1 cDNA transfection can inhibit the proliferation of K562 cells and colony formation, and promote its apoptosis; sense VEGF12 1 cDNA transfection is contrary to the results. The results of this study demonstrate that autocrine VEGF plays an important role in the proliferation and apoptosis of CML cell line K5 62