ABSTRACT
Treatment of human breast epithelial cells MCF-10F with 17-ß-estradiol has been reported to result in E2-transformed cells which have given rise to highly invasive C5 cells that in turn generate tumors in SCID mice. From these tumors, various cell lines, among which C5-A6-T6 and C5-A8-T8, were obtained. Although different phases of the tumorigenesis process in this model have been studied in molecular biology and image analysis assays, no cytological data on apoptotic ratios and mitotic abnormalities have been established to accompany the various steps leading to 17-ß-estradiol-treated MCF-10F cells to tumorigenesis. Here we detected that the apoptotic ratio decreases with the transformation and tumorigenesis progress, except for the tumor cell line C5-A8-T8, probably on account of its more intense proliferation rate and a more rapid culture medium consumption. Increased frequency of mitotic abnormalities contributed by triple- and tetrapolar metaphases, and by lagging chromosomes and chromosome bridges observed at the anaphase found by transformation and tumorigenesis progress. However, no difference was found under these terms when the C5-A6-T6 and C5-A8-T8 tumor cell lines were compared to each other. Present findings are in agreement with the nuclear instability and enrichment of dysregulated genes in the apoptotic process promoted by transformation and tumorigenesis in 17-ß-estradiol-treated MCF-10F cells.
Subject(s)
Apoptosis/drug effects , Breast Neoplasms/pathology , Epithelial Cells/drug effects , Epithelial Cells/pathology , Estradiol/pharmacology , Mitosis/drug effects , Animals , Cell Line, Transformed , Cell Line, Tumor , Cell Transformation, Neoplastic , Female , Humans , MiceABSTRACT
Treatment of human breast epithelial cells MCF-10F with 17-β-estradiol has been reported to result in E2-transformed cells which have given rise to highly invasive C5 cells that in turn generate tumors in SCID mice. From these tumors, various cell lines, among which C5-A6-T6 and C5-A8-T8, were obtained. Although different phases of the tumorigenesis process in this model have been studied in molecular biology and image analysis assays, no cytological data on apoptotic ratios and mitotic abnormalities have been established to accompany the various steps leading to 17-β-estradiol-treated MCF-10F cells to tumorigenesis. Here we detected that the apoptotic ratio decreases with the transformation and tumorigenesis progress, except for the tumor cell line C5-A8-T8, probably on account of its more intense proliferation rate and a more rapid culture medium consumption. Increased frequency of mitotic abnormalities contributed by triple- and tetrapolar metaphases, and by lagging chromosomes and chromosome bridges observed at the anaphase found by transformation and tumorigenesis progress. However, no difference was found under these terms when the C5-A6-T6 and C5-A8-T8 tumor cell lines were compared to each other. Present findings are in agreement with the nuclear instability and enrichment of dysregulated genes in the apoptotic process promoted by transformation and tumorigenesis in 17-β-estradiol-treated MCF-10F cells.
O tratamento das células epiteliais mamárias humanas MCF-10F com 17-β-estradiol tem sido relatado como resultando nas células transformadas E2, que deram origem às células C5, altamente invasivas, e que geraram tumores em camundongos SCID. A partir desses tumores foram originadas em cultura células tumorais, dentre as quais C5-A6-T6 e C5-A8-T8. Embora diversas fases do processo tumorigênico neste modelo tenham sido estudadas por ensaios de biologia molecular e análise de imagem, não foram ainda estimados dados citológicos referentes a índices apoptóticos e anomalias mitóticas que acompanhassem os vários passos que levam as células CF-10F tratadas com 17-β-estradiol à tumorigênese. Neste trabalho detectamos que o índice apoptótico decresce com a transformação e o avanço da tumorigênese, exceto na linhagem celular tumoral C5-A8-T8, provavelmente por causa de sua velocidade de proliferação mais intensa, que poderia levá-la a um consumo mais rápido do meio de cultura presente e à morte celular. Um aumento na frequência de anomalias mitóticas contribuídas por metáfases tripolares e tetrapolares e por pontes cromossômicas e cromossomos desgarrados, identificáveis na anáfase, foi observado com a transformação e o progresso da tumorigênese. Contudo não foram detectadas diferenças nesses parâmetros quando se compararam as linhagens tumorais C5-A6-T6 e C5-A8-T8 entre si. Os presentes achados estão de acordo com a instabilidade nuclear e o enriquecimento em desregulação de genes que atuam no processo apoptótico, promovidos pela transformação e tumorigênese nas células MCF-10F tratadas com 17-β-estradiol.
Subject(s)
Female , Humans , Animals , Mice , Apoptosis , Epithelial Cells , Epithelial Cells/pathology , Estradiol/pharmacology , Mitosis , Cell Line, Transformed , Cell Line, Tumor , Cell Transformation, NeoplasticABSTRACT
AIM: The purposes of this study were: 1) to evaluate age and gender differences in physical activity (PA) of children and adolescents; 2) to find out if children and adolescents fulfill the PA recommendations of 60 min x day(-1) of moderate (MPA) to vigorous PA (VPA). METHODS: PA was assessed in 265 female and 238 male subjects, ranging from 6 to 18 years of age, grouped in 4 age groups, with MTI ActiGraph model 7164, during 7 consecutive days. The MTI actigraph data was reduced to bouts (30-, 20-, 10-, and 5-min) and minutes spent in MPA, VPA, and very VPA (VVPA). RESULTS: The oldest boys and girls revealed a lower number of PA bouts than the younger ones. Significant gender differences were found in daily VPA, F(1, 492)=37.67, P<0.001; and VVPA F(1, 494)=24.11, P<0.001. Boys were more active than girls. Significant age group differences were also found in MPA, F(3, 494)=87.4, P<0.001; VPA, F(3, 492)=78.15, P<0.001; and VVPA, F(3, 454)=54.89, P<0.001. In both genders MPA, VPA and VVPA decreased with age. Till the age of 14, children had means between 79.6+/-30.6 and 144.1+/-76.9 min*day(-1) of PA. After this age, there was a decrease to 44.1+/-19.9 min*day(-1) in girls and to 56.3+/-31.9 min*day(-1) in boys. CONCLUSION: Boys had more minutes a day of VPA and VVPA than girls. PA decreased with age. The subjects of this study, aged 6 to 15, fulfilled the recommendations of 60 min x day(-1) of MPA to VPA.
