Oktem G.Sercan O.Guven U.Uslu R.Uysal A.Goksel G.Ayla S.Bilir A.2024-07-222024-07-2220141021335Xhttp://akademikarsiv.cbu.edu.tr:4000/handle/123456789/16993Cancer stem cells (CSCs) have the ability to self-renew similar to normal stem cells. This process is linked with metastasis and resistance to chemotherapy and radiotherapy. In the present study, we constructed an in vitro differentiation model for CSCs. CSCs isolated and proliferated for one passage were maintained as monolayers or spheroid-forming cells with serum included media for differentiation process. Differentiation of adhesion molecules and cellular ultrastructural properties were investigated and compared in both monolayer and spheroid cultures. CD133+/CD44+ cancer-initiating cells were isolated from DU-145 human prostate cancer cell line monolayer cultures and propagated as tumor spheroids and compared with the remaining heterogeneous cancer cell bulk population. Microarray-based gene expression analysis was applied to determine genes with differential expression and protein expression levels of candidates were analyzed by immunohistochemistry. Electron microscopy showed detailed analysis of morphology. TGFβ1 was found to be significantly upregulated in monolayer CSCs. High expression levels of VCAN, COL7A1, ITGβ3, MMP16, RPL13A, COL4A2 and TIMP1 and low expression levels of THBS1, MMP1 and MMP14 were detected when CSCs were maintained as serum-grown prostate CSC spheroids. Immunohistochemistry supported increased immunoreactivity of TGFβ1 in monolayer cultures and VCAN in spheroids. CSCs were found to possess multipotential differentiation capabilities through upregulation and/or downregulation of their markers. TGFβ1 is a triggering molecule, it stimulates versican, Col7A1, ITGβ3 and, most importantly, the upregulation of versican was only detected in CSCs. Our data support a model where CSCs must be engaged by one or more signaling cascades to differentiate and initiate tumor formation. This mechanism occurs with intracellular and extracellular signals and it is possible that CSCc themselves may be a source for extracellular signaling. These molecules functioning in tumor progression and differentiation may help develop targeted therapy.EnglishAll Open Access; Bronze Open AccessAntigens, CDAntigens, CD44Cell DifferentiationCell Line, TumorCollagen Type VIIGene Expression Regulation, NeoplasticGlycoproteinsHumansIntegrin beta3MaleNeoplastic Stem CellsPeptidesProstatic NeoplasmsSpheroids, CellularTransforming Growth Factor beta1Tumor Markers, BiologicalVersicansbeta3 integrincollagen type 7collagen type 7A1interstitial collagenasematrix metalloproteinase 14matrix metalloproteinase 16nerve cell adhesion moleculenerve cell adhesion molecule 1osteopontintransforming growth factor beta1unclassified druguvomorulinversicanbeta3 integrinCD133 antigenCD44 protein, humanCOL7A1 protein, humancollagen type 7glycoproteinHermes antigenITGB3 protein, humanleukocyte antigenpeptideTGFB1 protein, humantransforming growth factor beta1tumor markerVCAN protein, humanversicanarticlecancer stem cellcarcinogenesisCDH1 genecell differentiationcell proliferationCOL7A1 genecontrolled studydown regulationgene expressionhumanhuman cellimmunoreactivityin vitro studymaleMMP1 geneMMP16 genepriority journalprostate cancerprotein expressionsignal transductionTGF beta1 genetumor growthtumor spheroidupregulationVCAN genecancer stem cellcell differentiationgene expression regulationgeneticsmetabolismmulticellular spheroidpathologyprostate tumortumor cell lineArticle10.3892/or.2014.3252