Scott P A, Gleadle J M, Bicknell R, Harris A L
Molecular Oncology Group, ICRF, University of Oxford, John Radcliffe Hospital, UK.
Int J Cancer. 1998 Mar 2;75(5):706-12. doi: 10.1002/(sici)1097-0215(19980302)75:5<706::aid-ijc8>3.0.co;2-2.
Vascular endothelial growth factor (VEGF) is a potent angiogenic factor implicated in many pathological processes. We investigated the regulation of 4 alternatively spliced isoforms (121, 165, 189 and 206 amino acids) by hypoxia, hypoglycemia, acidity, female reproductive hormones and vitamin D in breast carcinoma cell lines representing different tumor phenotypes. There was a 17-fold difference in total VEGF mRNA expression across the cell lines. The isoform expression, 121 > 165 > 189, was unchanged by different culture conditions. Hypoxia was the most potent stimulus, and the cell lines demonstrated a 1.4- to 6.9-fold range of VEGF induction, maintained when other hypoxically regulated genes (phosphoglycerate kinase 1 and glucose transporter 1) and a HIF-1-dependent reporter gene were examined. The relative inducibility of the genes was maintained in each cell line, but basal expression was independent of -fold induction. VEGF expression decreased under acidic conditions in 2 cell lines, but the hypoxia stimulus remained effective under acidic conditions. Hypoglycemia, female reproductive hormones and vitamin D exerted no effect on expression, nor did inhibitors of mutant ras. Our results show that VEGF expression varies widely between cell lines and that capacity to respond to hypoxia is also cell specific, relating mostly to the hypoxic sensing of the cell and the signal transduction mechanism. Such characteristics, if maintained in vivo, have implications for the angiogenic potential of different tumor cells under normal and hypoxic conditions.
血管内皮生长因子(VEGF)是一种强效血管生成因子,与许多病理过程有关。我们研究了缺氧、低血糖、酸性环境、女性生殖激素和维生素D对代表不同肿瘤表型的乳腺癌细胞系中4种可变剪接异构体(分别含121、165、189和206个氨基酸)的调控作用。各细胞系中VEGF总mRNA表达存在17倍的差异。异构体表达为121>165>189,不受不同培养条件的影响。缺氧是最有效的刺激因素,各细胞系中VEGF的诱导倍数在1.4至6.9倍之间,在检测其他缺氧调控基因(磷酸甘油酸激酶1和葡萄糖转运蛋白1)以及HIF-1依赖性报告基因时仍保持这一倍数。各细胞系中基因的相对诱导能力保持不变,但基础表达与诱导倍数无关。在2种细胞系中,酸性条件下VEGF表达下降,但缺氧刺激在酸性条件下仍然有效。低血糖、女性生殖激素和维生素D对表达无影响,突变型ras的抑制剂也无作用。我们的结果表明,VEGF表达在不同细胞系之间差异很大,对缺氧的反应能力也是细胞特异性的,这主要与细胞的缺氧感知和信号转导机制有关。这些特征若在体内得以维持,则对不同肿瘤细胞在正常和缺氧条件下的血管生成潜力具有重要意义。