난소의 황체협막세포에서 E-cadherin, N-cadherin과세포부착에 미치는 Prostaglandin F2 Alpha의 영향 = Effect of Prostaglandin F2 Alpha on E-cadherin, N-cadherin and Cell Adhesion in Ovarian Luteal Theca Cells

다국어 초록 (Multilingual 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 (PGF2α) 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 (3β-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 PGF2α for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed 3β-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 PGF2α treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM PGF2α compared to the 0 mM PGF2α treatment groups (P＜0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM PGF2α treatment group than in the 0 mM PGF2α treatment group (P＜0.05). In conclusion, PGF2α 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.

국문 초록 (Abstract)

Cadherin은 원형질막에 존재하며 세포-세포 결합에 관여하며, 황체 구조 유지에 필수적인 단백질이다. 본 연구에서는prostaglandin F2 alpha (PGF2α)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 LTCs 로 판단하였다. 이 후 steroidogenic 기능 및 혈관세포 유무를판단하기 위해 3β-HSD 및 VEGF2R mRNA 발현을 확인하였으며, E-cadherin 및 N-cadherin mRNA를 사용하여 LTCs 내cadherin의 존재여부를 판단하였다. 또한 0, 10−5, 10−4 및10−3 M PGF2α를 24시간 동안 처리하여 LTCs의 E- 및N-cadherin 단백질을 관찰한 후 세포-세포 접착 실험을 실시하였다. 그 결과, LTCs에서 3β-HSD mRNA가 발현되었지만, VEGFR2 mRNA는 발현되지 않았으며, E-cadherin 및N-cadherin mRNA 모두 발현되는 것을 확인하였다. 또한 E- 및 N-cadherin 단백질은 10−5, 10−4 및 10−3 M PGF2α를 처리한 LTCs에서 응집되어 발현되는 것을 확인하였으며, PGF2α에의해 LTCs의 세포부착 효율이 유의적으로 감소된 것을 확인하였다. 결론적으로 PGF2α는 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.

참고문헌 (Reference)

• 1 Stocco C, "The molecular control of corpus luteum formation, function, and regression" 28 : 117-149, 2007
• 2 Mui KL, "The mechanical regulation of integrin– cadherin crosstalk organizes cells, signaling and forces" 129 : 1093-1100, 2016
• 3 Desouza M, "The actin cytoskeleton as a sensor and mediator of apoptosis" 2 : 75-87, 2012
• 4 Maroni D, "TGFB1 disrupts the angiogenic potential of microvascular endothelial cells of the corpus luteum" 124 : 2501-2510, 2011
• 5 Skarzynski DJ, "Role of tumor necrosis factor- and nitric oxide in luteolysis in cattle" 29 : 340-346, 2005
• 6 Yadav VK, "Prostaglandin F2-mediated activation of apoptotic signaling cascades in the corpus luteum during apoptosis" 280 : 10357-10367, 2005
• 7 Yuan W, "Programmed cell death in human ovary is a function of follicle and corpus luteum status" 82 : 3148-3155, 1997
• 8 Tajima K, "Ovarian theca cells in follicular function" 15 : 591-609, 2007
• 9 Okuda K, "Multiple roles of TNF super family members in corpus luteum function" 1 : 95-, 2003
• 10 Korzekwa A, "Luteolytic effect of prostaglandin F2 on bovine corpus luteum depends on cell composition and contact" 43 : 464-472, 2008
• 11 McCracken JA, "Luteolysis : a neuroendocrine-mediated event" 79 : 263-323, 1999
• 12 Brannian JD, "Isolation of ovine luteal cell subpopulations by flow cytometry" 48 : 495-502, 1993
• 13 Harald Herrmann, "Intermediate filaments: from cell architecture to nanomechanics" Springer Science and Business Media LLC 8 (8): 562-573, 2007
• 14 Korzekwa A, "Influence of prostaglandin F2 analogues on the secretory function of bovine luteal cells and ovarian arterial contractility in vitro" 199 : 131-137, 2014
• 15 Po JW, "Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of epithelial EpCAM and mesenchymal N-cadherin" 7 : 1-10, 2018
• 16 Firyal S. Khan-Dawood, "Immunocytochemical Localization and Expression of E-Cadherin, β-Catenin, and Plakoglobin in the Baboon (Papio anubis) Corpus Luteum1" Oxford University Press (OUP) 55 (55): 246-253, 1996
• 17 Skarzynski D, "Growth and regression in bovine corpora lutea : regulation by local survival and death pathways" 48 : 25-37, 2013
• 18 Grzesiak M, "Flutamide alters -catenin expression and distribution, and its interactions with E-cadherin in the porcine corpus luteum of mid-and late pregnancy" 30 : 1341-1352, 2015
• 19 Shirasuna K, "Expression of mRNA for cell adhesion molecules in the bovine corpus luteum during the estrous cycle and PGF2alpha-induced luteolysis" 53 : 1319-1328, 2007
• 20 김민성, "Expression of Fas and TNFR1 in the Luteal Cell Types Isolated from the Ovarian Corpus Luteum" 대한의생명과학회 25 (25): 107-112, 2019
• 21 Berisha B, "Expression and localization of gap junctional connexins 26 and 43 in bovine periovulatory follicles and in corpus luteum during different functional stages of oestrous cycle and pregnancy" 44 : 295-302, 2009
• 22 Anna KORZEKWA, "Effects of Prostaglandin F2α and Nitric Oxide on the Secretory Function of Bovine Luteal Cells" Japanese Society of Animal Reproduction 50 (50): 411-417, 2004
• 23 Abera A, "EP2 receptor mediated cAMP release is augmented by PGF2 activation of the FP receptor via the calcium-calmodulin pathway" 22 : 71-79, 2010
• 24 Sundfeldt K, "E-cadherin-catenin complex in the rat ovary : cell-specific expression during folliculogenesis and luteal formation" 118 : 375-386, 2000
• 25 Machell N, "E-and N-cadherin expression and distribution during luteinization in the rat ovary" 125 : 791-800, 2003
• 26 Baum B, "Dynamics of adherens junctions in epithelial establishment, maintenance, and remodeling" 192 : 907-917, 2011
• 27 Spanel-Borowski K, "Diversity of ultrastructure in different phenotypes of cultured microvessel endothelial cells isolated from bovine corpus luteum" 266 : 37-49, 1991
• 28 Alila H, "Control of progesterone production in small and large bovine luteal cells separated by flow cytometry" 82 : 645-655, 1988
• 29 Yamagishi-Kimura R, "Contribution of prostanoid FP receptor and prostaglandins in transient inflammatory ocular hypertension" 8 : 11098-, 2018
• 30 Berisha B, "Changes in the expression of prostaglandin family members in bovine corpus luteum during the estrous cycle and pregnancy" 7 : 622-634, 2018
• 31 O'shea J, "Cellular composition of the cyclic corpus luteum of the cow" 85 : 483-487, 1989
• 32 Groten T, "Cell junctional proteins in the human corpus luteum : changes during the normal cycle and after HCG treatment" 21 : 3096-3102, 2006
• 33 Sheetz MP, "Cell control by membrane–cytoskeleton adhesion" 2 : 392-396, 2001
• 34 Spitschak M, "Bovine large luteal cells show increasing de novo DNA methylation of the main ovarian CYP19A1 promoter P2" 178 : 37-45, 2012
• 35 Yadav VK, "Apoptosis during spontaneous and prostaglandin F2-induced luteal regression in the Buffalo Cow(Bubalus bubalis) : involvementof mitogenactivated protein kinases" 67 : 752-759, 2002
• 36 Yuyin Yi, "Activin A promotes ovarian cancer cell migration by suppressing E-cadherin expression" Elsevier BV 382 (382): 111471-, 2019
• 37 Alila H, "A comparison of the effects of cyclooxygenase prostanoids on progresterone production by small and large bovine luteal cells" 36 : 259-270, 1988