Does a tumor-associated antigen, EpCAM, regulate formation and function of tight junction in striated squamous epithelium?

2018 Fiscal Year Final Research Report

• Project/Area Number: 17H07304
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Morphological basic dentistry
• Research Institution: Fukuoka Dental College
• Principal Investigator: SEO AKIRA 福岡歯科大学, 口腔歯学部, 助教 (70804037)
• Research Collaborator: Inai Tetsuichiro , 教授 ; Kitagawa Norio ; Otani Takahito ; Nikaido Misaki ; Ozaki Akane ; Miyazono Shoji
• Project Period (FY): 2017-08-25 – 2019-03-31
• Keywords: タイト結合 / EpCAM / ケラチノサイト / 細胞間透過性

Outline of Final Research Achievements

In cancer, highly expressed EpCAM is a transmembrane glycoprotein involved in cell adhesion, cell proliferation, and tumor progression, and expression is also observed in stratified squamous epithelium, which is a normal tissue. We aimed to create keratinocytes from which the EpCAM gene has been disrupted and to investigate changes in tight junction formation and intercellular permeability during stratified squamous epithelium formation using a three-dimensional culture system. In this study, a three-dimensional culture method using the mouse keratinocyte strain (K38) formed a non-keratinized stratified squamous epithelium like the oral mucosa epithelium The expression of the tight junction membrane protein claudin (CL) 1, 4, 6, 7 was examined by Western blot and fluorescent immunostaining to confirm that it was almost similar to that of the living body. However, cells in which the EpCAM gene was disrupted could not be obtained.

Free Research Field

細胞生物

Academic Significance and Societal Importance of the Research Achievements

本研究では、線維芽細胞を使わずケラチノサイト単独で重層扁平上皮を構築する三次元培養系を用いることで、線維芽細胞からの間接的な影響（成長因子などの分泌蛋白）を排除して、上皮単独で解析できる点で意義がある。また、目的とする遺伝子産物をマウスで破壊して解析する場合、一般的には多大な労力と費用がかかる。さらに、機能的・形態的に明瞭な変化がなかったり、胎生致死であるためにその後の解析が困難な場合があり得る。本研究では、ケラチノサイトで働く遺伝子の機能を解析するためのひとつのスクリーニング系として活用できる点で意義がある。