Differential expression of CD44(S) and variant isoforms v3, v10 in three-dimensional cultures of mouse melanoma cell lines.
Multi-cellular spheroids (MCS) generated from tumor cells serve as excellent in vitro models for understanding the mechanisms of tumor progression and micro-metastasis. We have compared the expression of molecular markers with reference to their growth as conventional adherent monolayers (2-D) and anchorage independent cultures (3-D) using two mouse melanoma cell lines, B16F10 and Clone M3. The two cell lines differed in their ability to form spheroids with respect to their aggregation potential, with B16F10 forming large clusters compared to Clone M3. A panel of molecular markers comprising cell adhesion molecules, cyclin dependent kinase inhibitors and members of the cadherin-catenin complex were analyzed by flow cytometry in 2-D and 3-D cultures. There was a distinct difference in the patterns of expression of CD44(S) and variant isoforms v3, v10 in spheroids compared to cells grown as monolayers in both cell lines. Also, there was an increase in cells positive for CDK inhibitor p27 in 3-D cultures from the B16F10 cell line. The expression of alpha and gamma catenin was down regulated in spheroids. As these molecules are implicated in the regulation of cell proliferation, alterations in the expression of these molecules in 3-D cultures compared to their 2-D counterparts suggests the importance of spheroids as experimental model for tumorigenesis.
National Centre for Cell Science (NCCS), NCCS Complex, Ganeshkhind, Pune 411007, India. shiras99@hotmail.com
Anjali
A
Shiras
Arti
A
Bhosale
Anjali
A
Patekar
Varsha
V
Shepal
Padma
P
Shastry
D000818:Animals; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D015820:Cadherins / Q000032:analysis; D002448:Cell Adhesion; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000379:methods; D018797:Cell Cycle Proteins / Q000096:biosynthesis / Q000235:genetics; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050760:Cyclin-Dependent Kinase Inhibitor p27; D016213:Cyclins / Q000032:analysis; D003598:Cytoskeletal Proteins / Q000096:biosynthesis / Q000235:genetics; D051180:Desmoplakins; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D018960:Hyaluronan Receptors / Q000096:biosynthesis* / Q000235:genetics; D046911:Macromolecular Substances; D008546:Melanoma, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D020033:Protein Isoforms / Q000096:biosynthesis* / Q000235:genetics; D018874:Spheroids, Cellular / Q000378:metabolism / Q000473:pathology; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics; D051178:alpha Catenin; D051176:beta Catenin; D051185:gamma Catenin
D000951:Antigens, Neoplasm; C121450:CD44V3,8-10; C117722:CD44v10 antigen; C495267:CTNNB1 protein, mouse; D015820:Cadherins; C493957:Cdkn1a protein, mouse; C493961:Cdkn1b protein, mouse; D018797:Cell Cycle Proteins; C495272:Ctnna1 protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; D018960:Hyaluronan Receptors; C495293:Jup protein, mouse; D046911:Macromolecular Substances; D020033:Protein Isoforms; D015534:Trans-Activators; D025521:Tumor Suppressor Proteins; D051178:alpha Catenin; D051176:beta Catenin; D051185:gamma Catenin; D050760:Cyclin-Dependent Kinase Inhibitor p27