Effects of tumor microenviromental factors on VEGF expression

Wang,Feng; Xu,Ping; Xie,Kuang‑Chen; Chen,Xia‑Fang; Li,Chuan‑Yuan; Huang,Qian

doi:10.3892/br.2013.115

Effects of tumor microenviromental factors on VEGF expression

Authors:
- Feng Wang
- Ping Xu
- Kuang‑Chen Xie
- Xia‑Fang Chen
- Chuan‑Yuan Li
- Qian Huang
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Affiliations: Experimental Research Center, The First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, P.R. China, Department of Radiation Oncology, University of Colorado Health Sciences Center, Aurora, CO 80010, USA
Published online on: May 28, 2013 https://doi.org/10.3892/br.2013.115
Pages: 539-544

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Abstract

The expression of vascular endothelial growth factor (VEGF) is regulated by microenvironmental factors within the tumors, such as hypoxia, free radicals, pH imbalance and nutrient deficiency. The purpose of this study was to observe VEGF activity in tumor cells under different stress conditions. A plasmid was generated, consisting of green fluorescent protein (GFP) fused to a 1,217‑bp sequence, which was located downstream and upstream of the transcriptional start site of VEGF, respectively. The plasmid was stably transfected into 4T1 mouse breast carcinoma cells. Cells were cultured in a medium with nitric oxide (NO) donor sodium nitroprusside (SNP), hypoxia‑mimetic agent deferoxamine mesylate (DFX), H2O2, absence of serum and lowered or elevated pH, or were heat‑shocked, followed by measurement of VEGF activity by reverse transcription polymerase chain reaction (RT‑PCR) and Elisa. hypoxia, SNP and H2O2 led to increments of VEGF mRNA and protein expression, as well as of GFP expression. The pH alterations, serum deprivation and heat shock reduced VEGF mRNA expression, but had little effect on GFP expression. The results demonstrated that VEGF expression may be influenced by a number of microenvironmental factors and these factors may play important roles in regulating VEGF expression during tumorigenesis.

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1	Carmeliet P and Jain RK: Angiogenesis in cancer and other diseases. Nature. 407:249–257. 2000. View Article : Google Scholar : PubMed/NCBI
2	Folkman J: Tumor angiogenesis. Adv Cancer Res. 43:175–203. 1985. View Article : Google Scholar
3	Folkman J, Watson K, Ingber D and Hanahan D: Induction of angiogenesis during the transition from hyperplasia to neoplasia. Nature. 339:58–61. 1989. View Article : Google Scholar : PubMed/NCBI
4	Bellen JA, Van Diest PJ and Baak JP: Relationships between vascularization and proliferation in invasive breast cancer. J Pathol. 189:309–318. 1999. View Article : Google Scholar
5	Li CY, Shan S, Huang Q, Braun RD, Lanzen J, Hu K, Lin P and Dewhirst MW: Initial stages of tumor cell-induced angiogenesis: evaluation via skin window chambers in rodent models. J Natl Cancer Inst. 92:143–147. 2000. View Article : Google Scholar
6	de Vries C, Escobedo JA, Ueno H, Houck K, Ferrara N and William LT: The fms-like tyrosine kinase, a receptor for vascular endothelial growth factor. Science. 255:989–991. 1992.
7	Millauer B, Wizigmann-Voos S, Schnurch H, Martinez R, Møller NP, Risau W and Ullrich A: High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell. 72:835–846. 1993. View Article : Google Scholar : PubMed/NCBI
8	Mazure NM, Chen EY, Laderoute KR and Giaccia AJ: Induction of vascular endothelial growth factor by hypoxia is modulated by a phosphatidylinositol 3-kinase/Akt signaling pathway in Ha-ras-transformed cells through a hypoxia inducible factor-1 transcriptional element. Blood. 90:3322–3331. 1997.
9	Bolat F, Kayaselcuk F, Nursal TZ, Yagmurdur MC, Bal N and Demirhan B: Microvessel density, VEGF expression, and tumor-associated macrophages in breast tumors: correlations with prognostic parameters. J Exp Clin Cancer Res. 25:365–372. 2006.PubMed/NCBI
10	Holash J, Maisonpierre PC, Compton D, Boland P, Alexander CR, Zagzag D, Yancopoulos GD and Wiegand SJ: Vessel cooption, regression, and growth in tumors mediated by angiopoietins and VEGF. Science. 284:1994–1998. 1999. View Article : Google Scholar : PubMed/NCBI
11	Yancopoulos GD, Davis S, Gale NW, Rudge JS, Wiegand SJ and Holash J: Vascular-specific growth factors and blood vessel formation. Nature. 407:242–248. 2000. View Article : Google Scholar : PubMed/NCBI
12	Ikeda E, Achen MG, Breier G and Risau W: Hypoxia-induced transcriptional activation and increased mRNA stability of vascular endothelial growth factor in C6 glioma cells. J Biol Chem. 270:19761–19766. 1995. View Article : Google Scholar : PubMed/NCBI
13	Forsythe JA, Jiang BH, Iyer NV, Agani F, Leung SW, Koos RD and Semenza GL: Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol. 16:4604–4613. 1996.PubMed/NCBI
14	Levy NS, Goldberg MA and Levy AP: Sequencing of the human vascular endothelial growth factor (VEGF) 3′ untranslated region (UTR): conservation of five hypoxia-inducible RNA-protein binding sites. Biochem Biophys Acta. 1352:167–173. 1997.
15	Shima DT, Kuroki M, Deutsch U, Ng YS, Adamis AP and D’Amore PA: The mouse gene for vascular endothelial growth factor. Genomic structure, definition of the transcriptional unit, and characterization of transcriptional and post-transcriptional regulatory sequences. J Biol Chem. 271:3877–3883. 1996.
16	Fukumura D, Xu L, Chen Y, Gohongi T, Seed B and Jain RK: Hypoxia and acidosis independently up-regulate vascular endothelial growth factor transcription in brain tumors in vivo. Cancer Res. 61:6020–6024. 2001.PubMed/NCBI
17	Melillo G, Taylor LS, Brooks A, Musso T, Cox GW and Varesio L: Functional requirement of the hypoxia-responsive element in the activation of the inducible nitric oxide synthase promoter by the iron chelator desferrioxamine. J Biol Chem. 272:12236–12243. 1997. View Article : Google Scholar : PubMed/NCBI
18	Wang GL and Semenza GL: Desferrioxamine induces erythropoietin gene expression and hypoxia-inducible factor 1 DNA-binding activity: implications for models of hypoxia signal transduction. Blood. 82:3610–3615. 1993.PubMed/NCBI
19	Potier E, Ferreira E, Dennler S, Mauviel A, Oudina K, Logeart-Avramoglou D and Petite H: Desferrioxamine-driven upregulation of angiogenic factor expression by human bone marrow stromal cells. J Tissue Eng Regen Med. 2:272–278. 2008. View Article : Google Scholar : PubMed/NCBI
20	Koshikawa N, Takenaga K, Tagawa M and Sakiyama S: Therapeutic efficacy of the suicide gene driven by the promoter of vascular endothelial growth factor gene against hypoxic tumor cells. Cancer Res. 60:2936–2941. 2000.PubMed/NCBI
21	Liu LZ, Hu XW, Xia C, He J, Zhou Q, Shi X, Fang J and Jiang BH: Reactive oxygen species regulate epidermal growth factor-induced vascular endothelial growth factor and hypoxia-inducible factor-1α expression through activation of AKT and P70S6K1 in human ovarian cancer cells. Free Radic Biol Med. 41:1521–1533. 2006.PubMed/NCBI
22	Roybal CN, Hunsaker LA, Barbash O, Vander Jagt DL and Abcouwer SF: The oxidative stressor arsenite activates vascular endothelial growth factor mRNA transcription by an ATF4-dependent mechanism. J Biol Chem. 280:20331–20339. 2005. View Article : Google Scholar
23	Jung YD, Nakano K, Liu W, Gallick GE and Ellis LM: Extracellular signal-regulated kinase activation is required for up-regulation of vascular endothelial growth factor by serum starvation in human colon carcinoma cells. Cancer Res. 59:4804–4807. 1999.