VEGF upregulates VEGF receptor-2 on human outer root sheath cells and stimulates proliferation through ERK pathway

Springer Science and Business Media LLC - Tập 39 - Trang 8687-8694 - 2012
Wei Li1, Zhong-Fa Lu1, Xiao-Yong Man1, Chun-Ming Li2, Jiong Zhou1, Jia-Qi Chen1, Xiao-Hong Yang3, Xian-Jie Wu1, Sui-Qing Cai1, Min Zheng1
1Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
2Department of Dermatology, Second Affiliated Hospital, Nanchang University School of Medicine, Nanchang, China
3Department of Dermatology, Zhejiang Provincial Hospital of Traditional Chinese Medicine, Hangzhou, China

Tóm tắt

Vascular endothelial growth factor (VEGF) is a key regulator of physiological and pathological angiogenesis. The biological effects of VEGF are mediated by receptor tyrosine kinases. VEGF receptor-2, the primary receptor for VEGF, is thought to mediate most functional effects. In this study, we examined the expression and roles of VEGF receptor-2 on human outer root sheath cells (ORS). The expression of VEGFR-2 was determined at mRNA and protein levels by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Localization of VEGFR-2 in ORS cells was detected by immunofluorescence. The effect of VEGF on ORS cell proliferation was determined by MTT assays. Our data showed the expression of VEGFR-2 on ORS cells at both mRNA and protein levels. Immunostaining for VEGFR-2 demonstrated strong signal on cultured ORS cells. Exogenous VEGF165 stimulated proliferation of ORS cells and upregulated expression of VEGFR-2 in a dose-dependent manner. Moreover, VEGF165 induced phosphorylation of VEGFR-2, PLC-γ1, PKC-α, MEK, and p44/42 MAPK (ERK1/2) in a time-dependent manner. Taken together, human ORS cells express functional VEGF receptor-2 and exogenous VEGF165 upregulates expression of VEGFR-2 and stimulates proliferation of ORS cells via VEGFR-2 mediated ERK signaling pathway.

Tài liệu tham khảo

Paus R, Foitzik K (2004) In search of the “hair cycle clock”: a guided tour. Differentiation 72:489–511 Paus R, Cotsarelis G (1999) The biology of hair follicles. N Engl J Med 341:491–497 Stenn KS, Paus R (2001) Controls of hair follicle cycling. Physiol Rev 81:449–494 Pena JC, Kelekar A, Fuchs EV et al (1999) Manipulation of outer root sheath cell survival perturbs the hair-growth cycle. EMBO J 18:3596–3603 Roy H, Bhardwaj S, Yla-Herttuala S (2006) Biology of vascular endothelial growth factors. FEBS Lett 580:2879–2887 Ferrara N, Gerber HP, LeCouter J (2003) The biology of VEGF and its receptors. Nat Med 9:669–676 Holmes K, Roberts OL, Thomas AM et al (2007) Vascular endothelial growth factor receptor-2: structure, function, intracellular signalling and therapeutic inhibition. Cell Signal 19:2003–2012 Olsson AK, Dimberg A, Kreuger J et al (2006) VEGF receptor signalling-in control of vascular function. Nat Rev Mol Cell Biol 7:359–371 Weis SM, Cheresh DA (2005) Pathophysiological consequences of VEGF-induced vascular permeability. Nature 437:497–504 Man XY, Yang XH, Cai SQ et al (2006) Immunolocalization and expression of vascular endothelial growth factor receptors (VEGFRs) and neuropilins (NRPs) on keratinocytes in human epidermis. Mol Med 12:127–136 Yang XH, Man XY, Cai SQ et al (2006) Expression of VEGFR-2 on HaCaT cells is regulated by VEGF and plays an active role in mediating VEGF induced effects. Biochem Biophys Res Commun 349:31–38 Man XY, Yang XH, Cai SQ et al (2008) Overexpression of vascular endothelial growth factor (VEGF) receptors on keratinocytes in psoriasis: regulated by calcium independent of VEGF. J Cell Mol Med 12:649–660 Kozlowska U, Blume-Peytavi U, Kodelja V et al (1998) Expression of vascular endothelial growth factor (VEGF) in various compartments of the human hair follicle. Arch Dermatol Res 290:661–668 Man XY, Yang XH, Cai SQ et al (2009) Expression and localization of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 in human epidermal appendages: a comparison study by immunofluorescence. Clin Exp Dermatol 34:396–401 Yano K, Brown LF, Detmar M (2001) Control of hair growth and follicle size by VEGF-mediated angiogenesis. J Clin Invest 107:409–417 Millar SE (2002) Molecular mechanisms regulating hair follicle development. J Invest Dermatol 118:216–225 Rendl M, Lewis L, Fuchs E (2005) Molecular dissection of mesenchymal-epithelial interactions in the hair follicle. PLoS Biol 3:e331 Ancelin M, Chollet-Martin S, Herve MA et al (2004) Vascular endothelial growth factor VEGF189 induces human neutrophil chemotaxis in extravascular tissue via an autocrine amplification mechanism. Lab Invest 84:502–512 Fan F, Wey JS, McCarty MF et al (2005) Expression and function of vascular endothelial growth factor receptor-1 on human colorectal cancer cells. Oncogene 24:2647–2653 Kim EJ, Park HY, Yaar M et al (2005) Modulation of vascular endothelial growth factor receptors in melanocytes. Exp Dermatol 14:625–633 Cebe-Suarez S, Zehnder-Fjallman A, Ballmer-Hofer K (2006) The role of VEGF receptors in angiogenesis; complex partnerships. Cell Mol Life Sci 63:601–615 Takahashi T, Yamaguchi S, Chida K et al (2001) A single autophosphorylation site on KDR/Flk-1 is essential for VEGF-A-dependent activation of PLC-gamma and DNA synthesis in vascular endothelial cells. EMBO J 20:2768–2778 Takahashi T, Ueno H, Shibuya M (1999) VEGF activates protein kinase C-dependent, but Ras-independent Raf-MEK-MAP kinase pathway for DNA synthesis in primary endothelial cells. Oncogene 18:2221–2230 Wong C, Jin ZG (2005) Protein kinase C-dependent protein kinase D activation modulates ERK signal pathway and endothelial cell proliferation by vascular endothelial growth factor. J Biol Chem 280:33262–33269 Issbrucker K, Marti HH, Hippenstiel S et al (2003) p38 MAP kinase–a molecular switch between VEGF-induced angiogenesis and vascular hyperpermeability. FASEB J 17:262–264 Fujio Y, Walsh K (1999) Akt mediates cytoprotection of endothelial cells by vascular endothelial growth factor in an anchorage-dependent manner. J Biol Chem 274:16349–16354 Holmqvist K, Cross MJ, Rolny C et al (2004) The adaptor protein shb binds to tyrosine 1175 in vascular endothelial growth factor (VEGF) receptor-2 and regulates VEGF-dependent cellular migration. J Biol Chem 279:22267–22275 Lachgar S, Moukadiri H, Jonca F et al (1996) Vascular endothelial growth factor is an autocrine growth factor for hair dermal papilla cells. J Invest Dermatol 106:17–23 Goldman CK, Tsai JC, Soroceanu L et al (1995) Loss of vascular endothelial growth factor in human alopecia hair follicles. J Invest Dermatol 104:18S–20S Limat A, Mauri D, Hunziker T (1996) Successful treatment of chronic leg ulcers with epidermal equivalents generated from cultured autologous outer root sheath cells. J Invest Dermatol 107:128–135