Expression of Epithelial Cell Adhesion Molecule (EpCAM) in Tumor Spheroids of Human Colorectal Adenocarcinoma Cells
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Cell Technologies in Biology and Medicine, No. 3, November, 2020
Expression of Epithelial Cell Adhesion Molecule (EpCAM) in Tumor Spheroids of Human Colorectal Adenocarcinoma Cells A. M. Gisina1, Ya. S. Kim1, A. N. Gabashvili2, A. V. Tsvetkova1, I. V. Vakhrushev1, K. N. Yarygin1, and A. Yu. Lupatov1
Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 3, pp. 178-184, September, 2020 Original article submitted November 26, 2019 We studied the formation of spheroids by Caco-2, SW480, and HCT116 human colorectal adenocarcinoma cell lines under low-adhesion culturing conditions. Of these three cell lines, only HCT116 formed stable tumor spheroids. Flow cytometry analysis of 19 surface mar kers in monolayer HCT116 culture and spheroids formed by these cells revealed considerable similarity of the expression profiles in these two culturing modes. The only exception was EpCAM molecule: its expression in spheroids was 3-fold higher than in the monolayer culture. Scanning confocal laser microscopy showed equal EpCAM distribution in the inner mass of the spheroids. Key Words: tumor spheroids; colorectal cancer; HCT116; surface markers; EpCAM Tumor spheroids are 3D cultures of cancer cells formed during culturing under conditions providing minimum cell attachment and spreading on the substrate. It is believed that tumor spheroids are closer by their properties to real solid tumors and are more adequate model in comparison with monolayer (2D) cultures of tumor cells [8]. Four types of 3D cancer cell cultures are commonly used: 1) tumor spheroids obtained under non-adhesion conditions from stable cell lines; 2) tumor-spheres obtained under non-adhesion conditions from the primary culture of tumor cells in a serum-free medium containing growth factors and intended for culture enrichment with cancer stem cells; 3) tumor spheroids obtained by mechanical dissociation of the tumor tissue to exclude stroma cells; 4) organotypic spheroids obtained from mechanically minced tumor tissue and preserving the original cellular composition of the tumor. The most appropriate model is chosen based on the purpose of the study and the type of tumor [14]. 1 V. N. Orekhovich Research Institute of Biomedical Chemistry, Moscow, Russia; 2National University of Science and Technology MISIS, Moscow, Russia; Address for correspondence: alisa.gisina@gmail. com. A. M. Gisina
Tumor spheroids, in contrast to monolayer cultures, can partially reproduce differentiation of the original tumor. For instance, the outer layers of spheroids from hepatoma cell lines can differentiate into smooth densely packed polarized cells [11] and microvilli may appear on the surface of spheroid cells from intestinal cancer cell lines [4,16]. Histological signs of ovarian carcinomas that are absent in monolayer cultures can also reappear in tumor spheroids [10]. Some differences between 2D and 3D tumor cell cultures relate to increasing or decreasing the expression of surface molecules responsible for contacts with other cells and extracellular matrix.
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