RESUMO
BACKGROUND: Melanoma is one of the most aggressive and deadliest skin tumor. Cholesterol content in melanoma cells is elevated, and a portion of it accumulates into lipid rafts. Therefore, the plasma membrane cholesterol and its lateral organization might be directly linked with tumor development. ATP Binding Cassette A1 (ABCA1) transporter modulates physico-chemical properties of the plasma membrane by modifying cholesterol distribution. Several studies linked the activity of the transporter with a different outcome of tumor progression depending on which type. However, no direct link between human melanoma progression and ABCA1 activity has been reported yet. METHODS: An immunohistochemical study on the ABCA1 level in 110 patients-derived melanoma tumors was performed to investigate the potential association of the transporter with melanoma stage of progression and prognosis. Furthermore, proliferation, migration and invasion assays, extracellular-matrix degradation assay, immunochemistry on proteins involved in migration processes and a combination of biophysical microscopy analysis of the plasma membrane organization of Hs294T human melanoma wild type, control (scrambled), ABCA1 Knockout (ABCA1 KO) and ABCA1 chemically inactivated cells were used to study the impact of ABCA1 activity on human melanoma metastasis processes. RESULTS: The immunohistochemical analysis of clinical samples showed that high level of ABCA1 transporter in human melanoma is associated with a poor prognosis. Depletion or inhibition of ABCA1 impacts invasion capacities of aggressive melanoma cells. Loss of ABCA1 activity partially prevented cellular motility by affecting active focal adhesions formation via blocking clustering of phosphorylated focal adhesion kinases and active integrin ß3. Moreover, ABCA1 activity regulated the lateral organization of the plasma membrane in melanoma cells. Disrupting this organization, by increasing the content of cholesterol, also blocked active focal adhesion formation. CONCLUSION: Human melanoma cells reorganize their plasma membrane cholesterol content and organization via ABCA1 activity to promote motility processes and aggressiveness potential. Therefore, ABCA1 may contribute to tumor progression and poor prognosis, suggesting ABCA1 to be a potential metastatic marker in melanoma.
Assuntos
Melanoma , Humanos , Membrana Celular , Análise por Conglomerados , Transportador 1 de Cassete de Ligação de ATPRESUMO
BACKGROUND: Melanoma is one of the most aggressive and deadliest skin tumor. Cholesterol content in melanoma cells is elevated, and a portion of it accumulates into lipid rafts. Therefore, the plasma membrane cholesterol and its lateral organization might be directly linked with tumor development. ATP Binding Cassette A1 (ABCA1) transporter modulates physico-chemical properties of the plasma membrane by modifying cholesterol distribution. Several studies linked the activity of the transporter with a different outcome of tumor progression depending on which type. However, no direct link between human melanoma progression and ABCA1 activity has been reported yet. METHODS: An immunohistochemical study on the ABCA1 level in 110 patients-derived melanoma tumors was performed to investigate the potential association of the transporter with melanoma stage of progression and prognosis. Furthermore, proliferation, migration and invasion assays, extracellular-matrix degradation assay, immunochemistry on proteins involved in migration processes and a combination of biophysical microscopy analysis of the plasma membrane organization of Hs294T human melanoma wild type, control (scrambled), ABCA1 Knockout ( ABCA1 KO) and ABCA1 chemically inactivated cells were used to study the impact of ABCA1 activity on human melanoma metastasis processes. RESULTS: The immunohistochemical analysis of clinical samples showed that high level of ABCA1 transporter in human melanoma is associated with a poor prognosis. Depletion or inhibition ofABCA1 impacts invasion capacities of aggressive melanoma cells. Loss of ABCA1 activity partially prevented cellular motility by affecting active focal adhesions formation via blocking clustering of phosphorylated focal adhesion kinases and active integrin ß3. Moreover, ABCA1 activity regulated the lateral organization of the plasma membrane in melanoma cells. Disrupting this organization, by increasing the content of cholesterol, also blocked active focal adhesion formation. CONCLUSION: Human melanoma cells reorganize their plasma membrane cholesterol content and organization via ABCA1 activity to promote motility processes and aggressiveness potential. Therefore, ABCA1 may contribute to tumor progression and poor prognosis, suggesting ABCA1 to be a potential metastatic marker in melanoma.
Assuntos
Humanos , Melanoma , Análise por Conglomerados , Membrana Celular , Transportador 1 de Cassete de Ligação de ATPRESUMO
In this work was evaluated the cytotoxic activity of dehydrodieugenol B (1) and methyldehydrodieugenol B (2) isolated from Nectandra leucantha (Lauraceae) through cytokinesis-block micronucleus (CBMN) and Comet assay. Compounds 1 and 2 displayed in vitro toxicity against human melanoma cells (SKMEL-147) with IC50 values of 4.4 and 43.6⯵g/mL, respectively. The interaction of these compounds with resistant human melanoma cell line SKMEL-29 was also investigated. Obtained results showed a concentration-response relationship for DNA damage (DI and D%) in SKMEL-29 cells for compounds 1 and 2 causing an increase in DNA damage on their lower concentrations. Concerning the cytokinesis-block micronucleus cytome (CBMNCyt) assay, all treatments demonstrated an increase in cytostatic and cytotoxic indexes. In micronucleus quantification, compound 1 displayed higher index in comparison to control. Regarding necrotic and apoptotic cells, treatments with SKMEL-29 cells demonstrated 100% of cell death induced by compounds 1 and 2 at 25 and 88⯵g/mL, respectively. Additionally, it was observed that apoptosis is prevalent in SKMEL-147 cells treated with compound 1, while necrotic cells were observed in SKMEL-29 cells treated with both compounds. In conclusion, compounds 1 and 2 are suggested as promising cytotoxic agents against human melanoma resistant cells, emphasizing the potential use of these neolignans for the treatment of melanoma.
Assuntos
Citotoxinas/farmacologia , Lauraceae , Lignanas/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ensaio Cometa , Dano ao DNA , Humanos , Melanoma/tratamento farmacológico , Testes para Micronúcleos , Caules de PlantaRESUMO
Vasculogenic mimicry process has generated great interest over the past decade. So far, however, there have been only a few matrices available that allow us to study that process in vitro. Here, we have developed an innovative hydrogel platform with defined composition that mimics the structural architecture and biological functions of the extracellular matrix for vasculogenic mimicry of human melanoma cells (SK-MEL-28). We chemically immobilized IKVAV peptide on bacterial nanocellulose (BNC) fibers. BNC-IKVAV hydrogel was found to improve the adhesion and proliferation of SK-MEL-28 cells on the top and bottom surfaces. Particularly, the bottom surface of BNC-IKVAV induced SK-MEL-28 cells to organize themselves as well-established networks related to the vasculogenic mimicry process. Finally, our results showed that not only BNC-IKVAV but also BNC hydrogels can potentially be used as a three-dimensional platform that allows the screening of antitumor drugs. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2741-2749, 2018.
Assuntos
Bactérias/química , Adesão Celular , Proliferação de Células , Celulose/química , Hidrogéis/química , Laminina/química , Melanoma , Nanoestruturas/química , Neovascularização Patológica , Fragmentos de Peptídeos/química , Animais , Linhagem Celular Tumoral , Humanos , Melanoma/sangue , Melanoma/metabolismo , Melanoma/patologia , Camundongos , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologiaRESUMO
AIM: To evaluate the diagnostic value of dynamic contrast-enhanced computed tomography (CT) and magnetic resonance imaging (MRI) in the differential diagnosis of hepatic angiomyolipoma (HAML) and hepatocellular carcinoma (HCC) and to clarify the relationship between histopathological features and CT or MRI imaging performances in HAML. MATERIAL AND METHODS: Six HAML and 33 non-cirrhotic HCC patients confirmed by histopathology were retrospectively analyzed. The serum biomarkers, CT and MRI examinations were conventionally performed before the confirmatory histological diagnosis. The clinical data from their medical records was also analyzed. RESULTS: Six HAML patients were annotated as two types according to CT and MRI imaging characteristics, including hypovascular type (n = 1) and hypervascular type (n = 5). The imaging performances of the 33 HCC patients were hypervascular type. Moreover, all the 5 hypervascular type HAML patients were misdiagnosed as HCC by CT or MRI. We also found that the hypervascular type of HAML patients contained more vessels and less fatty tissues in histopathology than hypovascular type of HAML patients. However, the clinical features included HCC high risk factors (hepatitis B or C), non-specific symptoms, male and increased serum alpha fetoprotein (AFP) were more common in HCC patients than HAML patients (P < 0.05, respectively). CONCLUSIONS: The CT or MRI imaging performances of HAML patients containing more vessels and less fatty tissues in histopathology resemble the imaging performance of HCC patients. These clinical features may be of great help in the differential diagnosis in the current clinical practices.
Assuntos
Angiomiolipoma/diagnóstico por imagem , Angiomiolipoma/patologia , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/patologia , Imageamento por Ressonância Magnética , Tomografia Computadorizada por Raios X , Adulto , Idoso , Angiomiolipoma/química , Biomarcadores Tumorais/análise , Biópsia , Carcinoma Hepatocelular/diagnóstico por imagem , Carcinoma Hepatocelular/patologia , Meios de Contraste/administração & dosagem , Diagnóstico Diferencial , Erros de Diagnóstico , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/química , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Retrospectivos , Fatores de RiscoRESUMO
Cancer is a public health problem and the second leading cause of death worldwide. The incidence of cutaneous melanoma has been notably increasing, resulting in high aggressiveness and poor survival rates. Taking into account the antitumor activity of biflorin, a substance isolated from Capraria biflora L. roots that is cytotoxic in vitro and in vivo, this study aimed to demonstrate the action of biflorin against three established human melanoma cell lines that recapitulate the molecular landscape of the disease in terms of genetic alterations and mutations, such as the TP53, NRAS and BRAF genes. The results presented here indicate that biflorin reduces the viability of melanoma cell lines by DNA interactions. Biflorin causes single and double DNA strand breaks, consequently inhibiting cell cycle progression, replication and DNA repair and promoting apoptosis. Our data suggest that biflorin could be considered as a future therapeutic option for managing melanoma.
Assuntos
DNA/metabolismo , Melanoma/genética , Naftoquinonas/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Metilases de Modificação do DNA/genética , Enzimas Reparadoras do DNA/genética , GTP Fosfo-Hidrolases/genética , Humanos , Proteínas de Membrana/genética , Mutação , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas B-raf/genética , Timidilato Sintase/genética , Proteína Supressora de Tumor p53/genética , Proteínas Supressoras de Tumor/genética , Proteínas ras/genéticaRESUMO
Background Polycosanols derived from plant species have traditionally been used in medicine as antiproliferative agents for treating various viruses (primarily the herpes simplex virus). However, few studies have studied their effects on hyperproliferative cell lines. In this work, the antiproliferative capacity of polycosanols from tall-oil pitch, obtained from black liquor soaps in the kraft pulping process of cellulose (specifically from Pinus radiata, Pinus taede, and Eucalyptus globulus), was evaluated on CHO-K1 and CRL-1974 human melanoma cell lines. Results The proliferative capacities and cell viabilities were measured for 72 and 140 h, respectively. Treatment with docosanol produced differential effects on the CHO-K1 and human melanoma cells and significantly affected their proliferation rates, but not their cell viabilities. Tetracosanol produced a significant negative effect on the proliferation of human melanoma cells, and this effect was less than that caused by docosanol. However, it had no effect on the proliferation of CHO-K1 cells and did not induce any significant effect on the viability of the studied cell lines. Conclusion Docosanol and tetracosanol induced antiproliferative effects on the studied cell lines and exhibited significantly greater effects on the oncogenic cell lines. Prior to this study, the capacity of these polycosanols has never been investigated. Future studies will be necessary to determine their mechanisms of action on these cell systems.