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Title: Characterisation of KIM 2 murine mammary epithelial cells and reversion of the transformed phenotype via Tiam1 : a model for suppression of transformation in breast cancer?
Author: Kurian, Kathreena Mary
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2000
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The KIM2 mammary cell line, containing a temperature dependent SV40 large T antigen, was used as a model for normal and transformed mammary epithelial cells. At 37°C the mammary cell line appears epithelial and showed mammary differentiation (NKIM2), whereas at 33°C the cell line appears spindle-shaped and transformed (TKIM2). The aim of this study was therefore to determine the biological features of the NKIM2 and TKIM2 lines, revert the transformed TKIM2 to a non-transformed phenotype by retrovirally transducing Tiam 1 into the cell lines, assess the reverted cells in terms of the originally defined biological features, and explore new targets in the signalling pathway leading to phenotype reversion. The NKIM2 cell line showed features of epithelial differentiation including morphology, E-cadherin and ZO-1 expression, junctions and polarisation. In addition, the specialised myoepithelial cell type was identified among cultures. The NKIM2 cells also showed mammary differentiation in terms of milk protein expression in response to lactogenic hormones. However, the significance of the formation of dome-like structures in response to lactogenic stimuli remains unclear. The TKIM2 line demonstrated all the accepted features of transformation including morphology, lack of contact inhibition or serum dependence, anchorage independence in soft agar and tumour formation in scid mice. Unexpectedly, the NKIM2 lines also formed tumours in scid mice. This may have been a consequence of SV40 large T antigen binding p53 and resulting in a variable chromosomal content in both NKIM2 and TKIM2. There was no difference in the chromosomal locus for p53 and SEN6, which have been implicated in transformation and immortalisation respectively, in both cell lines. Retroviral transduction of control and active Tiam 1 constructs into both NKIM2 and TKIM2 lines was successful, with Tiam 1 protein expression in transduced cell lines at least three months after transduction.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (M.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available