Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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Effect of Prostaglandin F2 Alpha on E-cadherin, N-cadherin and Cell Adhesion in Ovarian Luteal Theca Cells

난소의 황체협막세포에서 E-cadherin, N-cadherin과 세포부착에 미치는 Prostaglandin F2 Alpha의 영향

  • Lee, Sang-Hee (Discipline of ICT, School of Technology, Environments and Design, University of Tasmania) ;
  • Jung, Bae Dong (College of Veterinary Medicine, Kangwon National University) ;
  • Lee, Seunghyung (College of Animal Life Sciences, Kangwon National University)
  • 이상희 (타즈매니아대학교 기술환경디자인대학 ICT학과) ;
  • 정배동 (강원대학교 수의과대학) ;
  • 이승형 (강원대학교 동물생명과학대학)
  • Received : 2019.07.19
  • Accepted : 2019.08.21
  • Published : 2019.09.30
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Abstract

Cadherins are essential transmembrane proteins that promote cell-cell adhesion and maintain the corpus luteum structure in the ovary. This study examined the influence of prostaglandin F2 alpha ($PGF2{\alpha}$) on E-cadherin, N-cadherin, and adhesion in luteal theca cells (LTCs). The luteal cells were isolated from the mid-phase corpus luteum, and the LTCs were cultured separately from the luteal heterogeneous cells according to the morphology of the mesenchymal cells and to determine if steroidogenic and endothelial cells of LTCs, 3beta-hydroxysteroid dehydrogenase ($3{\beta}$-HSD), and vascular endothelial growth factor receptor 2 (VEGFR2) mRNA were used. The LTCs were then incubated in the culture medium supplemented with 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed $3{\beta}$-HSD mRNA expression in the LTC but no VEGF2R mRNA expression. The E-cadherin and N-cadherin proteins of the LTCs were damaged in the 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM $PGF2{\alpha}$ compared to the 0 mM $PGF2{\alpha}$ treatment groups (P<0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM $PGF2{\alpha}$ treatment group than in the 0 mM $PGF2{\alpha}$ treatment group (P<0.05). In conclusion, $PGF2{\alpha}$ affected the disruption of cadherin proteins and cell adhesion in LTCs. These results may help better understand the cadherin and adhesion mechanism during corpus luteum regression in the ovary.

Cadherin은 원형질막에 존재하며 세포-세포 결합에 관여하며, 황체 구조 유지에 필수적인 단백질이다. 본 연구에서는 prostaglandin F2 alpha ($PGF2{\alpha}$)가 황체의 협막세포(luteal theca cells, LTCs)의 E-cadherin, N-cadherin 및 세포-세포부착에 미치는 영향에 대해서 수행하였다. 황체세포는 소의 황체중기 조직으로부터 분리하였으며, 황체세포 중에서 mesenchymal 세포 형태학적 특성을 가지는 세포만을 분리하여 LTCs로 판단하였다. 이 후 steroidogenic 기능 및 혈관세포 유무를 판단하기 위해 $3{\beta}$-HSD 및 VEGF2R mRNA 발현을 확인하였으며, E-cadherin 및 N-cadherin mRNA를 사용하여 LTCs 내 cadherin의 존재여부를 판단하였다. 또한 0, $10^{-5}$, $10^{-4}$ 및 $10^{-3}M$ $PGF2{\alpha}$를 24시간 동안 처리하여 LTCs의 E- 및 N-cadherin 단백질을 관찰한 후 세포-세포 접착 실험을 실시하였다. 그 결과, LTCs에서 $3{\beta}$-HSD mRNA가 발현되었지만, VEGFR2 mRNA는 발현되지 않았으며, E-cadherin 및 N-cadherin mRNA 모두 발현되는 것을 확인하였다. 또한 E-및 N-cadherin 단백질은 $10^{-5}$, $10^{-4}$ 및 $10^{-3}M$ $PGF2{\alpha}$를 처리한 LTCs에서 응집되어 발현되는 것을 확인하였으며, $PGF2{\alpha}$에 의해 LTCs의 세포부착 효율이 유의적으로 감소된 것을 확인하였다. 결론적으로 $PGF2{\alpha}$는 LTCs의 E- 및 N-cadherin을 붕괴시켜 세포부착을 감소시켰고, 이러한 결과는 황체퇴행의 새로운 원인을 밝혀 내기 위한 cadherin과 세포부착의 역할을 이해하는데 중요한 자료로 활용될 것으로 판단된다.

Keywords

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