Microfilaments, microtubules, and intermediate filaments form the cytoskeleton. These substructures play a significant role in cell motility, transport, divisions, differentiation, tumor transformation, and apoptosis. These processes are related with changes in cell shape, in which cytoskeletal proteins take an active part. In non-muscle cells, actin is an essential constituent of microfilaments, tubulin forms microtubules, and vimentin is one of the characteristic proteins of intermediate filaments. The differentiation of cells is associated inseparably with tissue and organ formation, and the induction of malignant cell differentiation can be a method of treatment, especially in hematopoietic steam cell disease therapy. In studies on tumor cell differentiation, agents such as cytokines, retinoids, forbol esters, and vitamin D3 are the most commonly used, and results show these substances may participate in different pathways of signal transduction. Retinoids and vitamin D3 mostly affect gene transcription via nuclear receptors, whereas cytokines act through membrane receptors. The results of studies show actin, tubulin, and vimentin reorganization during the differentiation of leukemia cells, but it remains unknown whether the observed changes are the cause or the result of the differentiation process.