Functional study of Gap Junctional Intercellular Communication in Human Keratinocyte Cell Line

Gap junctions, unique membrane structures that mediate the intercellular communications among almost all tissues, are composed of connexins in vertebrate. The essential role of gap junction-mediated intercellular communications(GJIC) for coordination of cell homeostasis, tissue functions and organ development are well reported by many researches using genetics and mutant studies. Mutations of connexin 26 and connexin 30 have been linked to deafness. In human middle ear cholesteatoma, upregulated expression of both connexin 43 and connexin 26 were observed compared to those in normal human skin. In this study, spontaneously immortalized non-tumorigenic and a highly diffenrentiating keratinocyte(HaCaT) cell line was used to find substances that can change the GJIC function via fuctional study using scrape-loading dye transfer(SLDT) method. Toxic drug level of acetic acid , H₂O₂, dexamethasone, Epicatechin epigallocatechin-3-gallate, Epicatechin and retinoic acid were assesed by Neutral red uptake test. The adequate concentration of these six material was determined and added to the culture medium of the HaCaT cell line. Scrape-loading dye transfer method was used to compare the functional aspect of GJIC in the different condition, By this method, fluorescent Lucifer yellow dye was introduced into the HaCaT cells and observed by the fluorescence inverted microscope. Captured images were analysed with image software and as a result, acetic acid, hydrogen peroxide, dexamethasone have down-regulated and epicatechin have up-regulated the GJIC function showing statistically significant difference(p<0.05) of the mean diffusion length of Lucifer yellow dye compared to that of the control group. The discovery of these substances will lead to the success of the medical treatment of cholesteatoma.

Author dc:creator

• 강, 희선

Contributors dc:contributor

• 정, 연훈
• 대학원 의학과
• 105161

• cholesteatoma
• HaCaT
• gap junction
• connexin
• scrape-loading and dye transfer
• acetic acid
• H2O2
• epicatechin
• epicatechin epigallocatechin-3-gallate
• retinoic acid

Language dc:language

ko

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000002076
000000002076

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/1636