Navegando por Palavras-chave "focal adhesion"
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- ItemAcesso aberto (Open Access)Estudo do efeito do EcTI (Enterolobium Contortisiliquum Trypsin Inhibitor), inibidor de metalo-proteases com ação no complexo de adesão focal, em células extraídas de pacientes com leiomioma uterino - análise comparativa com 17b-estradiol e progesterona(Universidade Federal de São Paulo (UNIFESP), 2016-01-28) Bonazza, Camila [UNIFESP]; Castro, Rodrigo de Aquino [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Introduction: Leiomyomas are common benign tumors of the female reproductive tract. Nowadays the surgery treatment is public health is the most effective. A striking feature of uterine leiomyomas is their dependency on ovarian steroids. In this scenario, the ability to test new drugs to modulate its effects prompted us to test the protease inhibitor, ECTI, extracted from Enteolobium plant contortisiliquum as a modulator of this condition, since ECTI proved to be an effective antitumor agent in several established lines of cancer. Thus, we evaluated the ECTI action on the focal adhesion pathway, targeting proteins Src / FAK and p130Cas and as a result, we investigated proteins crucial processes cycle and cell death. Methods: We used the primary culture of cells extracted myometrial women with leiomyoma and their adjacent tissues were submitted to total hysterectomy. The experiments were conducted using the 17?-estradiol (E2) and progesterone (P4) alone and concomitantly the ECTI. Results: In hormone therapy observed similar results to those observed in the clinic. We note that the presence of E2 leiomyoma cells did not show its modified metabolic activity, unlike observed increased cell proliferation by phosphorylation of ERK1 / 2, acting in the S phase of the cell cycle. Already P4 in turn, operates in cell cycle arrest and G1 / S with the participation of f-Rb and p21; increased proliferation (activity of Erk1 / 2). The combined hormone therapy, E2 + P4 showed antagonistic effect to that observed in isolated therapy, altering the metabolic activity of the myometrial cells leiomyoma with f-Rb expression, Bax activation and the activation of Akt (survival pathway of cell) myoblasts of the tumor surrounding tissue. Treatment with ECTI significantly alter the metabolic activity of myometrial leiomyoma cells but not in cells extracted from the tissue adjacent to the tumor. We note that the effect was potentiated in the presence of type I collagen surface and in parallel, hormonal therapy E2 and P4 also showed similar effect. Combination therapy with ECTI and steroid hormones proved the most effective, reducing drastically the metabolic activity of cells extracted leiomyoma, there was also participation p130Cas adhesion protein phosphorylation of residue Y410 on the tissue cells adjacent, and the residue Y165 cells of patients with leiomyoma. The cell cycle arrest involves increased expression of p21 and Rb-f. With the analysis of the cell death pathway, the protein is overexpressed presented. With regard to the pro-apoptotic proteins (Bax) and anti-apoptotic (Bcl-2 and Bcl-xL), in tissue, there is an elevated expression of Bax and Bcl xL in the presence of P4. However, when combining the ECTI the expression of Bax and Bcl-xL drops dramatically. Conclusion: These findings suggest that combination therapy ECTI with E2 + P4 changes the metabolic activity of cells of the uterine leiomyoma, with the participation of focal adhesion protein p130Cas, survival pathway by Akt, cell cycle arrest by f-Rb and p21 . I saw death there is the participation of pro-apoptotic protein Bax and in consideration of the antiapoptotic protein Bcl-xL. We can say that there is a synergism between the therapy and ECTI in the treatment of uterine leiomyoma cells. These results appear promising since, in the tissue adjacent the effect of ECTI cells and hormone therapy did not affect cell viability.
- ItemSomente MetadadadosEvaluation of photodynamic therapy in adhesion protein expression(Spandidos Publ Ltd, 2014-08-01) Pacheco-Soares, Cristina; Maftou-Costa, Maira [UNIFESP]; Da Cunha Menezes Costa, Carolina Genuncio; De Siqueira Silva, Andreza Cristina; Moraes, Karen C. M.; Univ Vale do Paraiba; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)Photodynamic therapy (PDT) is a treatment modality that has clinical applications in both non-neoplastic and neoplastic diseases. PDT involves a light-sensitive compound (photosensitizer), light and molecular oxygen. This procedure may lead to several different cellular responses, including cell death. Alterations in the attachment of cancer cells to the substratum and to each other are important consequences of photodynamic treatment. PDT may lead to changes in the expression of cellular adhesion structure and cytoskeleton integrity, which are key factors in decreasing tumor metastatic potential. HEp-2 cells were photosensitized with aluminum phthalocyanine tetrasulfonate and zinc phthalocyanine, and the proteins beta 1-integrin and focal adhesion kinase (FAK) were assayed using fluorescence microscopy. the verification of expression changes in the genes for FAK and beta 1 integrin were performed by reverse transcription-polymerase chain reaction (RT-PCR). the results revealed that HEp-2 cells do not express beta-integrin or FAK 12 h following PDT. It was concluded that the PDT reduces the adhesive ability of HEp-2 cells, inhibiting their metastatic potential. the present study aimed to analyze the changes in the expression and organization of cellular adhesion elements and the subsequent metastatic potential of HEp-2 cells following PDT treatment.
- ItemAcesso aberto (Open Access)Heparan sulfate proteoglycans: structure, protein interactions and cell signaling(Academia Brasileira de Ciências, 2009-09-01) Dreyfuss, Juliana Luporini [UNIFESP]; Regatieri, Caio Vinicius Saito [UNIFESP]; Jarrouge, Thais R. [UNIFESP]; Cavalheiro, Renan Pelluzzi [UNIFESP]; Sampaio, Lucia O. [UNIFESP]; Nader, Helena Bonciani [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Heparan sulfate proteoglycans are ubiquitously found at the cell surface and extracellular matrix in all the animal species. This review will focus on the structural characteristics of the heparan sulfate proteoglycans related to protein interactions leading to cell signaling. The heparan sulfate chains due to their vast structural diversity are able to bind and interact with a wide variety of proteins, such as growth factors, chemokines, morphogens, extracellular matrix components, enzymes, among others. There is a specificity directing the interactions of heparan sulfates and target proteins, regarding both the fine structure of the polysaccharide chain as well precise protein motifs. Heparan sulfates play a role in cellular signaling either as receptor or co-receptor for different ligands, and the activation of downstream pathways is related to phosphorylation of different cytosolic proteins either directly or involving cytoskeleton interactions leading to gene regulation. The role of the heparan sulfate proteoglycans in cellular signaling and endocytic uptake pathways is also discussed.