RESUMEN
Data are accumulating to support a role for adipose-derived mesenchymal stem cells (MSCs) in breast cancer progression; however, to date most studies have relied on adipose MSCs from non-breast sources. There is a particular need to investigate the role of adipose MSCs in the pathogenesis of basal-like breast cancer, which develops at a disproportionate rate in pre-menopausal African-American women with a gain in adiposity. The aim of this study was to better understand how breast adipose MSCs (bMSCs) contribute to the progression of basal-like breast cancers by relying on isogenic HMT-3255 S3 (pre-invasive) and T4-2 (invasive) human cells that upon transplantation into nude mice resemble this tumor subtype. In vitro results suggested that bMSCs may contribute to breast cancer progression in multiple ways. bMSCs readily penetrate extracellular matrix components in part through their expression of matrix metalloproteinases 1 and 3, promote the invasion of T4-2 cells and efficiently chemoattract endothelial cells via a bFGF-independent, VEGF-A-dependent manner. As mixed xenografts, bMSCs stimulated the growth, invasion and desmoplasia of T4-2 tumors, yet these resident stem cells showed no observable effect on the progression of pre-invasive S3 cells. While bMSCs form vessel-like structures within Matrigel both in vitro and in vivo and chemoattract endothelial cells, there appeared to be no difference between T4-2/bMSC mixed xenografts and T4-2 xenografts with regard to intra- or peri-tumoral vascularity. Collectively, our data suggest that bMSCs may contribute to the progression of basal-like breast cancers by stimulating growth and invasion but not vasculogenesis or angiogenesis.
Asunto(s)
Tejido Adiposo/patología , Neoplasias de la Mama/patología , Glándulas Mamarias Humanas/patología , Células Madre Mesenquimatosas/patología , Neoplasias Basocelulares/patología , Animales , Antígenos CD/metabolismo , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/patología , Neoplasias de la Mama/irrigación sanguínea , Línea Celular Tumoral , Quimiotaxis , Femenino , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Células Endoteliales de la Vena Umbilical Humana/fisiología , Humanos , Metaloproteinasas de la Matriz Secretadas/metabolismo , Células Madre Mesenquimatosas/metabolismo , Ratones , Ratones Desnudos , Invasividad Neoplásica , Trasplante de Neoplasias , Neoplasias Basocelulares/irrigación sanguínea , Carga Tumoral , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
A crucial step in human breast cancer progression is the acquisition of invasiveness. There is a distinct lack of human cell culture models to study the transition from preinvasive to invasive phenotype as it may occur "spontaneously" in vivo. To delineate molecular alterations important for this transition, we isolated human breast epithelial cell lines that showed partial loss of tissue polarity in three-dimensional reconstituted basement membrane cultures. These cells remained noninvasive; however, unlike their nonmalignant counterparts, they exhibited a high propensity to acquire invasiveness through basement membrane in culture. The genomic aberrations and gene expression profiles of the cells in this model showed a high degree of similarity to primary breast tumor profiles. The xenograft tumors formed by the cell lines in three different microenvironments in nude mice displayed metaplastic phenotypes, including squamous and basal characteristics, with invasive cells exhibiting features of higher-grade tumors. To find functionally significant changes in transition from preinvasive to invasive phenotype, we performed attribute profile clustering analysis on the list of genes differentially expressed between preinvasive and invasive cells. We found integral membrane proteins, transcription factors, kinases, transport molecules, and chemokines to be highly represented. In addition, expression of matrix metalloproteinases MMP9, MMP13, MMP15, and MMP17 was up-regulated in the invasive cells. Using small interfering RNA-based approaches, we found these MMPs to be required for the invasive phenotype. This model provides a new tool for dissection of mechanisms by which preinvasive breast cells could acquire invasiveness in a metaplastic context.
Asunto(s)
Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Transformación Celular Neoplásica , Regulación Neoplásica de la Expresión Génica , Animales , Línea Celular Tumoral , Femenino , Perfilación de la Expresión Génica , Humanos , Metaplasia/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Modelos Biológicos , Invasividad Neoplásica , Metástasis de la NeoplasiaRESUMEN
Polo-like kinase 1 (PLK1) has important functions in maintaining genome stability via its role in mitosis. Because PLK1 is up-regulated in many invasive carcinomas, we asked whether it may also play a role in acquisition of invasiveness, a crucial step in transition to malignancy. In a model of metaplastic basal-like breast carcinoma progression, we found that PLK1 expression is necessary but not sufficient to induce invasiveness through laminin-rich extracellular matrix. PLK1 mediates invasion via vimentin and beta1 integrin, both of which are necessary. We observed that PLK1 phosphorylates vimentin on Ser82, which in turn regulates cell surface levels of beta1 integrin. We found PLK1 to be also highly expressed in preinvasive in situ carcinomas of the breast. These results support a role for the involvement of PLK1 in the invasion process and point to this pathway as a potential therapeutic target for preinvasive and invasive breast carcinoma treatment.
Asunto(s)
Neoplasias de la Mama/enzimología , Proteínas de Ciclo Celular/metabolismo , Matriz Extracelular/enzimología , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Animales , Apoptosis/fisiología , Western Blotting , Neoplasias de la Mama/genética , Neoplasias de la Mama/terapia , Carcinoma Ductal de Mama/enzimología , Carcinoma Ductal de Mama/patología , Carcinoma Intraductal no Infiltrante/enzimología , Carcinoma Intraductal no Infiltrante/patología , Proteínas de Ciclo Celular/antagonistas & inhibidores , Proteínas de Ciclo Celular/genética , Movimiento Celular , Colágeno , Combinación de Medicamentos , Femenino , Humanos , Etiquetado Corte-Fin in Situ , Integrina beta1/metabolismo , Laminina/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Fosforilación , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/genética , Proteoglicanos , Proteínas Proto-Oncogénicas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas/genética , ARN Interferente Pequeño/farmacología , Vimentina/metabolismo , Quinasa Tipo Polo 1RESUMEN
Extracellular matrix (ECM) regulates cell morphology and gene expression in vivo; these relationships are maintained in three-dimensional (3D) cultures of mammary epithelial cells. In the presence of laminin-rich ECM (lrECM), mammary epithelial cells round up and undergo global histone deacetylation, a process critical for their functional differentiation. However, it remains unclear whether lrECM-dependent cell rounding and global histone deacetylation are indeed part of a common physical-biochemical pathway. Using 3D cultures as well as nonadhesive and micropatterned substrata, here we showed that the cell 'rounding' caused by lrECM was sufficient to induce deacetylation of histones H3 and H4 in the absence of biochemical cues. Microarray and confocal analysis demonstrated that this deacetylation in 3D culture is associated with a global increase in chromatin condensation and a reduction in gene expression. Whereas cells cultured on plastic substrata formed prominent stress fibers, cells grown in 3D lrECM or on micropatterns lacked these structures. Disruption of the actin cytoskeleton with cytochalasin D phenocopied the lrECM-induced cell rounding and histone deacetylation. These results reveal a novel link between ECM-controlled cell shape and chromatin structure and suggest that this link is mediated by changes in the actin cytoskeleton.
Asunto(s)
Forma de la Célula , Regulación de la Expresión Génica , Histonas/metabolismo , Glándulas Mamarias Humanas/citología , Acetilación , Actinas/metabolismo , Técnicas de Cultivo de Célula , Línea Celular , Cromatina/química , Cromatina/metabolismo , Citoesqueleto/metabolismo , Matriz Extracelular/metabolismo , Femenino , Perfilación de la Expresión Génica , Humanos , Conformación de Ácido Nucleico , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
Progression from normal to malignant phenotype involves aberrations in the reciprocal interactions of multiple cell types with each other and with other components of the microenvironment. In this issue of Cancer Cell, demonstrate that progression to breast cancer involves genotypic as well as gene expression changes that are cell type-specific, suggesting that targeted therapies delivered to the tumor may need to include drugs targeted not only to the tumor cells, but also to the other cell types in the tumor microenvironment.
Asunto(s)
Mama/metabolismo , Homeostasis , Mama/patología , Neoplasias de la Mama/genética , Transformación Celular Neoplásica/genética , Epitelio/metabolismo , Epitelio/patología , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica/fisiología , HumanosAsunto(s)
Mama/patología , Células Epiteliales/patología , Animales , Neoplasias de la Mama/patología , Células Epiteliales/metabolismo , Epitelio , Laminina/metabolismo , Microscopía Electrónica , Microscopía Fluorescente , Modelos Biológicos , Proteínas Nucleares , Fenotipo , Receptores de EstrógenosRESUMEN
The Annual Meeting of the American Society for Cell Biology (ASCB) is a diverse conference covering all topics in cell biology. While all of the basic biology presented at this meeting may potentially contribute to breast cancer research, there were a significant number of presentations and posters directly pertinent to this field. Here we have summarized the research that is of greatest immediate relevance to breast cancer, with particular emphasis on mammary gland development and tumorigenesis in vivo, three-dimensional in vitro models of mammary morphogenesis, alterations of signal transduction pathways in breast cancer, and global studies in expression profiling and drug screening.
Asunto(s)
Fenómenos Fisiológicos Celulares , Biología Molecular , Animales , Mama/citología , Mama/enzimología , Mama/crecimiento & desarrollo , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Citocinas/fisiología , Células Epiteliales/efectos de los fármacos , Matriz Extracelular/fisiología , Genes Supresores de Tumor/fisiología , Humanos , Glándulas Mamarias Animales/citología , Glándulas Mamarias Animales/enzimología , Glándulas Mamarias Animales/crecimiento & desarrollo , Metaloproteinasas de la Matriz/fisiología , Glicoproteínas de Membrana/farmacología , Modelos Biológicos , Neoplasias/etiología , Oncogenes/fisiología , Transducción de Señal/fisiología , Sociedades Científicas , Células Madre/enzimología , Células Madre/fisiología , Sintaxina 1 , Estados UnidosRESUMEN
The current paradigm for cancer initiation and progression rests on the groundbreaking discoveries of oncogenes and tumor suppressor genes. This framework has revealed much about the role of genetic alterations in the underlying signaling pathways central to normal cellular function and to tumor progression. However, it is clear that single gene theories or even sequential acquisition of mutations underestimate the nature of the genetic and epigenetic changes in tumors, and do not account for the observation that many cancer susceptibility genes (e.g. BRCA1, APC) show a high degree of tissue specificity in their association with neoplastic transformation. Therefore, the cellular and tissue context itself must confer additional and crucial information necessary for mutated genes to exert their influence. A considerable body of evidence now shows that cell-cell and cell-extracellular matrix (ECM) interactions are essential organizing principles that help define the nature of the tissue context, and play a crucial role in regulating homeostasis and tissue specificity. How this context determines functional integrity, and how its loss can lead to malignancy, appears to have much to do with tissue structure and polarity.