Suppression subtractive hybridisation to isolate differentially expressed genes involved in invasiveness of melanoma cell line cultured under different conditions
| Autor: | Pavol Kudela, Jaromir Pastorek, Maria Regendova, Vladimir Majerciak, Dagmar Eleveld-Trancikova, V. Zelnik, Silvia Pastorekova, Jozef Bizik |
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| Rok vydání: | 2002 |
| Předmět: |
Cancer Research
RHOA DNA Complementary Cell Polymerase Chain Reaction Proto-Oncogene Proteins p21(ras) Gene expression medicine Tumor Cells Cultured Humans Neoplasm Invasiveness Cloning Molecular Gene Melanoma Oncogene biology Reverse Transcriptase Polymerase Chain Reaction Membrane Proteins Nucleic Acid Hybridization Sequence Analysis DNA Cell cycle medicine.disease Molecular biology Immunohistochemistry Fibronectins Fibronectin Isoenzymes medicine.anatomical_structure Oncology Genetic Techniques Cyclooxygenase 2 Prostaglandin-Endoperoxide Synthases Cancer research biology.protein rhoA GTP-Binding Protein |
| Zdroj: | International journal of oncology. 20(3) |
| ISSN: | 1019-6439 |
| Popis: | We have previously analysed the invasion capacity of different melanoma cell lines in the three-dimensional dermal equivalent. The melanoma cell line M4Beu acquired invasive behaviour upon changing its cultivation conditions before the seeding on top of the collagen lattice from single cell suspension to spheroid. Based on this phenomenon SSH was used to search for the genes related to the invasive phenotype of melanoma cells. From differentially expressed clones we focused on four: fibronectin, RhoA, COXII, and H-ras-like protein. By RT-PCR the expression of these genes were tested in different populations (monolayer, spheroids on dermal equivalent) of melanoma cell line M4Beu and three additional melanoma cell lines. The expression of fibronectin was also examined by immunohistochemistry staining of co-culture spheroids-dermal equivalent. |
| Databáze: | OpenAIRE |
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