Knowing the mechanisms responsible for tissue differentiation and development of the organism is a major challenge both to developmental biology and molecular oncology. Coordinated behavior and cell movement are necessary for morphogenesis, wound healing and tumorigenesis. Change of the shape that is dependent on the cytoskeleton is the characteristic of the cells that move or differentiate. Here we investigated importance of the cytoskeleton in the growth and differentiation of MDCKII (Madin-Darby canine kidney epithelial cell) circular colonies and the differences between the two types of cells that emerged on the third day of growth. Compacted central cells increase phosphorylation of eIF2α which indicates increased cellular stress. eIF2α and phosphorylated eIF2α were detected by the Western blot analysis. By the same method we analyzed the level of cytoskeletal (actin, β-tubulin, vimentin, vinkulin) and adhesion proteins (β-catenin, E cadherin). We have shown that two types of cells differ significantly in the expression of vimentin, β catenin and E cadherin. Circular colony cells lost a differentiated appearance because of disruption of actin and microtubules by chemical compounds and suppresion of the vimentin expression by siRNA. Expression of nesprin – protein linking the cellular and nuclear skeleton – was analyzed via PCR method at the level of the messenger RNA. Disruption of actin and vimentin leads to a change in expression of nesprin 1 and nesprin 3. The results of this research point out the differences between the two types of cells inside circular colonies and the importance of cytoskeletal integrity in the growth and differentiation of the model tissue.