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How various critical oncogenic signals affect the differentiation state and thus affect the breast cancer subgrouping is still not well known.

The reason for this is partly the lack of in vitro culture systems, modeling both normal breast stem cell biology and neoplastic transformation.

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Dysregulation of the signaling pathways activated by the epidermal growth factor receptor (EGFR) family is oncogenic in many epithelial cells.

Amplification of the HER2 (Erb B2) gene is especially injurious in breast epithelium where upregulation of HER2 is seen in approximately 20% of all breast cancers, New breast cancer treatment regimens have, however, actualized new clinical challenges.

D492 is a breast epithelial cell line with stem cell properties that can undergo epithelial to mesenchyme transition (EMT), generate luminal- and myoepithelial cells and form complex branching structures in three-dimensional (3D) culture.

Here, we show that overexpression of HER2 in D492 (D492) was not oncogenic in vivo.

D492 is a breast epithelial cell line with stem cell properties that was established by isolating and immortalizing a MUC1 D492 cells differentiate into both luminal- and myoepithelial cells and form branching bi-layered cellular structures resembling the terminal duct lobular units when cultured in reconstituted basement membrane matrix (r BM).

Furthermore, these cells can respond to microenvironmental signals to undergo epithelial to mesenchyme transition (EMT).Initially, we analyzed HER2 and EGFR expression in the normal breast.CK19 and CK14 were used to identify luminal epithelial- and myoepithelial cells, respectively (Figure 1, top left).Co-staining of EGFR and HER2 with either CK19 or CK14 revealed distinct expression patterns with EGFR expression associated with the basal/myoepithelial compartment.HER2 expression was predominantly associated with the luminal epithelial cells (Figure 1, lower right).To analyze the differentiation- and oncogenic potential of HER2 on mammary progenitor cells, the protein was overexpressed in D492 (Supplementary Figure S1).

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