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

Mervenur Kalender,Muhammet Bulbul,Ilknur Keskin,Bircan Kolbasi

doi:10.31557/apjcp.2022.23.1.39

Mervenur Kalender, Muhammet Bulbul + Show 2 more

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https://doi.org/10.31557/apjcp.2022.23.1.39

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Abstract

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 for understanding the communication between cells and the in vivo microenvironment. Changes in expression levels of E-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 no expression of E-cadherin in endothelial cells. Methods:In our study, both two-dimensional and three dimensional mono-culture and co-culture models were created using Huvec and Ishikawa cells (endometrial cancer cell lines) to better 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-culture models were measured by immunofluorescence staining. Spheroid images were recorded using a Z-stack. Intensity measurements in both two-dimensional and three-dimensional mono-culture and co-culture models were made using the 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 significantly larger 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 Huvec and co-culture cells. Likewise, while the expression levels of β-catenin were higher in the mono-culture of Ishikawa cells 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 reduced in co-cultures of Ishikawa cells with Huvec cells in both two and three dimensions . These results support the idea that endothelial cells may cause changes in endometrial cancer progression by suppressing E-cadherin expression in Ishikawa cells.

Highlights

In vitro cell cultures are often used the mechanisms underlying in vivo cell behavior
Changes in expression levels of E-cadherin are closely related to cancer metastasis and progression. β-catenin mediates cell adhesion of E-cadherin
E-cadherin and β-catenin expression levels of two-dimensional and three dimensional mono-culture and co-culture models were measured by immunofluorescence staining

Summary

Introduction

In vitro cell cultures are often used the mechanisms underlying in vivo cell behavior. Two-dimensional cell culture techniques are preferred over in vivo studies due to their simplicity and high cell viability (Lee et al, 2008). Some evidence shows that two dimensional cell culture techniques cannot mimic the in vivo responses against the cellular or environmental changes. The morphology of cells that are isolated from the tissue and transferred to two dimensional cell culture conditions is different from the in vivo. Because of the implementation of unusual geometric and mechanical constraints to cells, two dimensional cell cultures have only some of the characteristics of normal tissues (Sun et al, 2006)

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