In 2D and 3D Cell Culture Models, Effects of Endothelial Cells on E-cadherin / β-catenin Expression Levels and Spheroid Sizes in Ishikawa Cells


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Kalender M., Bulbul M. V., KOLBAŞI B., KESKİN İ.

Asian Pacific Journal of Cancer Prevention, cilt.23, sa.1, ss.39-51, 2022 (Scopus) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 23 Sayı: 1
  • Basım Tarihi: 2022
  • Doi Numarası: 10.31557/apjcp.2022.23.1.39
  • Dergi Adı: Asian Pacific Journal of Cancer Prevention
  • Derginin Tarandığı İndeksler: Scopus, CAB Abstracts, EMBASE, MEDLINE, Veterinary Science Database, Directory of Open Access Journals
  • Sayfa Sayıları: ss.39-51
  • Anahtar Kelimeler: Co-culture, E-cadherin, Immunofluorescence, Spheroid, Β-catenin
  • İstanbul Medipol Üniversitesi Adresli: Evet

Özet

Objective: Increasing evidence shows that three dimensional cell culture models better reflect the in vivo tumor microenvironment than two dimensional cell culture models. Co-culture models are ideal cell culture models forunderstanding the communication between cells and the in vivo microenvironment. Changes in expression levels ofE-cadherin are closely related to cancer metastasis and progression. β-catenin mediates cell adhesion of E-cadherin.Endothelial cells are stromal cell components in the tumor microenvironment. It is known that there is little or noexpression of E-cadherin in endothelial cells. Methods: In our study, both two-dimensional and three dimensionalmono-culture and co-culture models were created using Huvec and Ishikawa cells (endometrial cancer cell lines) tobetter reflect cell interactions. Spheroids were followed for three days in the three-dimensional cell culture model.E-cadherin and β-catenin expression levels of two-dimensional and three dimensional mono-culture and co-culturemodels were measured by immunofluorescence staining. Spheroid images were recorded using a Z-stack. Intensitymeasurements in both two-dimensional and three-dimensional mono-culture and co-culture models were made usingthe Image J software. Study groups were evaluated by one-way analysis of variance (One-Way ANOVA). Values of p<0.05 were considered statistically significant. Results: The size of the co-culture spheroids was recorded significantlylarger than the mono-culture spheroids (p <0.0001). In two (p = 0.0175) and three dimensional models (p <0.0001),expression levels of E-cadherin in the mono-culture of Ishikawa cells were recorded significantly higher than in Huvecand co-culture cells. Likewise, while the expression levels of β-catenin were higher in the mono-culture of Ishikawacells in two-dimensional models (p <0.05), no significant difference was observed in three dimensional models (p>0.05). Conclusion: In summary, it has been noted that the expression levels of E-cadherin are significantly reducedin co-cultures of Ishikawa cells with Huvec cells in both two and three dimensions. These results support the ideathat endothelial cells may cause changes in endometrial cancer progression by suppressing E-cadherin expression inIshikawa cells