3-D vascularized breast cancer model to study the role of osteoblast in formation of a pre-metastatic niche

Breast cancer cells (BCCs) preferentially metastasize to bone. It is known that BCCs remotely primes the distant bone site prior to metastasis. However, the reciprocal influence of bone cells on the primary tumor is relatively overlooked. Here, to study the bone-tumor paracrine influence, a tri-cellular 3-D vascularized breast cancer tissue (VBCTs) model is engineered which comprised MDA-MB231, a triple-negative breast cancer cells (TNBC), fibroblasts, and endothelial cells. This is indirectly co-cultured with osteoblasts (OBs), thereby constituting a complex quad-cellular tumor progression model. VBCTs alone and in conjunction with OBs led to abnormal vasculature and reduced vessel density but enhanced VEGF production. A total of 1476 significantly upregulated and 775 downregulated genes are identified in the VBCTs exposed to OBs. HSP90N, CYCS, RPS27A, and EGFR are recognized as upregulated hub-genes. Kaplan Meier plot shows HSP90N to have a significant outcome in TNBC patient survivability. Furthermore, compared to cancer tissues without vessels, gene analysis recognized 1278 significantly upregulated and 566 downregulated genes in VBCTs. DKK1, CXCL13, C3 protein and BMP4 are identified to be downregulated hub genes in VBCTs. Together, a multi-cellular breast cancer model and culture protocols are established to study pre-metastatic events in the presence of OBs.

Changes in the morphological, immunological and genetic characteristics of Mel Ibr melanoma cells in response to low concentration of embryo calf serum

Objective: the purpose of the study is research of the changes in phenotypic and genetic characteristics of melanoma cells Mel Ibr cultured in growth medium with low concentration (5 %) of embryo calf serum. Materials and methods. Cell line Меl Ibr was pre-cultured in RPMI-1640 medium with 5 % FBS for 3passages, and then 200-400 cells were replaced on the Petri dish with a diameter of 60 mm and left in the incubator for 14 days to form colonies. Colonies of spindle form cells were cultivated in the same conditions for more than 30 passages. Cells were stained according to the method of Leishman to investigate the morphology. The expression of antigens was determined in the reaction of indirect immunofluorescence. Results. Exposure of Mel Ibr human melanoma cells to low concentration of embryo calf serum resulted in the appearance of a subclone with morphological, immunological and cytogenetical characteristics differed to those of parental cells. The subclone cells distinguished with spindle form, more small size, large nucleus which broadcasted on whole cytoplasm, formation spheroid and sharply reduced percentage of HLA-DR (+) and CD54 (+) cells, a significantly elevated percentage of CD63 (+) cells, and appearance of CD133 (+). The subclone karyotype differed to those of parental cells. Conclusoin. The subclone cells were characterized by the same features as stem tumor cells and could be use as tumor progression model.

Inferring a Tumor Progression Model for Neuroblastoma From Genomic Data

Purpose The knowledge of the key genomic events that are causal to cancer development and progression not only is invaluable for our understanding of cancer biology but also may have a direct clinical impact. The task of deciphering a model of tumor progression by requiring that it explains (or at least does not contradict) known clinical and molecular evidence can be very demanding, particularly for cancers with complex patterns of clinical and molecular evidence. Materials and Methods We formalize the process of model inference and show how a progression model for neuroblastoma (NB) can be inferred from genomic data. The core idea of our method is to translate the model of clonal cancer evolution to mathematical testable rules of inheritance. Seventy-eight NB samples in stages 1, 4S, and 4 were analyzed with array-based comparative genomic hybridization. Results The pattern of recurrent genomic alterations in NB is strongly stage dependent and it is possible to identify traces of tumor progression in this type of data. Conclusion A tumor progression model for neuroblastoma is inferred, which is in agreement with clinical evidence, explains part of the heterogeneity of the clinical behavior observed for NB, and is compatible with existing empirical models of NB progression.