The Effect of Nerve Growth Factor on Cell Proliferation and Expression of Its Receptors in Cultured Human Keratinocytes.
- Author:
Ji Ho RYOU
1
;
Tuk Woo LIM
;
Jai Kyung PARK
;
Nack In KIM
Author Information
1. Department of Dermatology, College of Medicine, Kyung Hee University, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Nerve growth factor;
Keratinocyte
- MeSH:
Antibodies;
Blotting, Northern;
Blotting, Western;
Cell Count;
Cell Proliferation*;
Humans*;
Immunohistochemistry;
Keratinocytes*;
Nerve Growth Factor*;
Nervous System;
Neurons;
RNA, Messenger
- From:Korean Journal of Dermatology
2001;39(2):161-167
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Nerve growth factor(NGF) is a neurotrophic polypeptide necessary for the survival and growth of some central neurons, as well as sensory afferent and sympathetic neurons. In addition to its actions on the nervous system, it also has a significant biologic effects on cells of the immune-inflammatory compartment. Recent studies suggest that NGF is an important autocrine growth factor and survival factor for keratinocytes which express both high- and low-affinity receptors for NGF. OBJECTIVE: The purpose of this study is to detect NFG receptors on cultured human keratinocytes and to evaluate the effect of NGF on proliferation of cultured human keratinocytes. METHODS: Cultured human keratinocytes were examined for the expression of high affinity receptor TrkA and low affinity receptor p75 by Northern blot, Western blot and immunocytochemistry. The effects of NGF on proliferation of cultured human keratinocytes were also evaluated. To specify the NGF effect on proliferation of human keratinocytes, excess of anti-NGF neutralizing polyclonal antibody was added. RESULTS: 1) NGF significantly stimulated the proliferation of keratinocytes in both 1% of keratinocyte growth supplement(KGS)-added medium(100ng/ml) and 0.2% KGS-added media(50, 100, 500ng/ml), (p<0.05). The cell number was dose-dependently increased in 0.2% KGS-added media. 2) Whenever we added 500 ng/ml of anti-NGF polyclonal antibody to the growth media, the cell number was statistically higher in 100ng/ml NGF-added group of 1% KGS-added medium, but there was not any statistical significance in 0.2% KGS-added media group. 3) Immunocytochemical staining with specific antibodies to TrkA and p75 revealed positive findings for these receptors, but TrkB and TrkC were not detected. 4) We could not detect both the mRNA and protein of TrkA and p75 by Northern and Western blot methods. CONCLUSION: These results suggest that both high affinity- and low affinity receptors for NGF are expressed in cultured human keratinocytes and NGF can induce keratinocyte proliferation.
