ABSTRACT
Background: Breast cancer has been one of the most common types of cancer, as the leading cause of women death in world. Breast cancer has known as a heterogenic disease that the clinical path in different patients would be very different. Since the current classification has not covered the diverse clinical course of breast cancer, lots of efforts has done to find new biological markers. Integrins are hetero dimmer proteins of alpha and beta subunits on cell membrane. After binding to extra cellular matrix [ECM], integrins activate MAPK pathway that regulated different activities like survival, differentiation, migration, immunologic response. The interaction of integrins and ECM have a key role in cancer cell activities like survival and metastasis
Objectives: In this study the expression alpha[v]beta[3] integrin, substrate-dependent morphology and ERK and p-ERK activation was compared in MCF7 and Hek-293 cells lines
Materials and Methods: The expression alpha[v]beta[3] integrin was assayed by flow cytometry. These cell lines were cultured on pre-covered plates with fibronectin [FN], fibrinogen [Fg] or collagen [Col] and the expression of ERK and p-ERK proteins was assessed in attached and free cells for each substrate after 1 hour incubation. The morphology of the cells have examined under an inverted phase contrast microscope at 15 min, 1 hour, 3 hours, 5 hours and 1 day of incubation
Results: Different substrate induced the expression ERK or p-ERK differently in the two cell lines. In MCF7 cells, substrates induced the expression of ERK in all the attached cells but free cells in BSA, collagen and Fg showed a lower expression of ERK. In comparison with Hek-293 cells althought all the attached cells have expressed ERK peotein but only free cells in collagen plates showed the expression of ERK. None of the cell lines has shown any expression of ERK and p-ERK in attached or free cells except for the Hek-293 free cells in collagen platees that have shown a weak signal for p-ERK
Conclusions: Overall the breast cancer cell lines MCF7 and Hek-293 cells have differently responded on similar substrates regarding morphology or ERK and MEK expressions
Subject(s)
Humans , Female , MCF-7 Cells , Extracellular Signal-Regulated MAP Kinases , Mitogen-Activated Protein Kinases , HEK293 Cells , Cells, Cultured , Cell Culture TechniquesABSTRACT
Members of the eukaryotic Hsp90 family function as important molecular chaperones in the assembly, folding and activation of cellular signaling in development. Two hsp90 genes, hsp90alpha and hsp90beta, have been identified in fish and homeothermic vertebrates but not in poikilothermic vertebrates. In the present study, the expression of hsp90alpha and hsp90beta genes in Xenopus laevis, which is phylogenetically positioned between zebrafish and mammals, has been addressed. Partial Xenopus hsp90alph and hsp90beta cDNA were identified and isolated using RT-PCR, and a full-length Xenopus hsp90beta cDNA was isolated from an embryonic cDNA library. Northern-blot analysis was used to study the expression of hsp90alpha and hsp90 beta genes in total RNA of the embryos and in situ hybridization was used to compare the expression of these genes with that of hsp70 and MyoD genes in Xenopus embryogenesis. Northern-blot analysis revealed that the hsp90fl gene was strongly expressed constitutively at all stages of embryogenesis, but weakly induced following the heat shock. In contrast, the hsp90alpha gene was weakly expressed in embryos at control temperature, but strongly up-regulated following heat shock. In situ hybridization results showed that hsp90a gene was observed predominantly in cells of the developing somite. Microscopic sections showed that hsp90alpha and MyoD mRNA are expressed in similar regions in somite and this pattern was distinct from that of hsp70 and hsp90beta. These data support the hypothesis that the presence of hsp90alpha and hsp90beta genes is conserved among vertebrates, and these genes are differentially regulated in a tissue, stress, and development stage-specific manner