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Title: Endometrial cancer cell lines as a model for metastasis
Author: Parkes-Reed, C. M.
ISNI:       0000 0004 7656 526X
Awarding Body: University of Liverpool
Current Institution: University of Liverpool
Date of Award: 2018
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Aim: To assess the hormone receptors (Androgen, Progesterone, Oestrogen alpha and Oestrogen beta) and metastasis inducing protein (MIP; AGR2, S100A4, S100P) expression and location in endometrial cancer (EC) cell lines. To then modulate MIP expression using hormone treatments (17β-estradiol (E2), DHT and MPA), and shRNA technology, measuring the impact on the phenotype and functionality of the cells with regard to their proliferation, migration and chemoresistance. Method: Using four EC cell lines (Ishikawa, HEC1A, RL95-2 and MFE280) MIP expression was detected using immunoblotting (immunocytochemistry, immunofluorescence and western blotting) techniques. Cells lines were treated with hormones to modulate AGR2 and S100P expression. Function of S100P in the RL95-2 cell line was achieved by transduction with S100p shRNA in lentiviral plasmids. Cell lines were grown in the chorioallantoic membrane (CAM) model to assess the tumour formation capabilities of the cells along with the effect of chemotherapeutic drugs on the cell line tumours. Results: A pragmatic workflow for ECresearch was created to guide cell line projects for future basic research. The cell lines have differing growth patterns in 2D culture and as such different methods were needed to assess the less well differentiated RL95-2 and MFE280 cell lines. MIPS were expressed in all four cell lines in varying degrees and in differing subcellular compartments. THE HEC1A and RL95-2 cell lines were able to grow tumours in the CAM assay whereas ISK cells just grew on the CAM surface. S100P knockdown is possible in the RL95-2 cell line and knockdown resulted in significant reduction in S100p transcript and protein levels. The knockdown cells had reduced proliferation, migration capabilities and increased chemoresistance to doxorubicin and paclitaxel determined by MTT and FACS analyses. Conclusions: This panel of four cell lines offer a range of cell lines to begin ECresearch investigations. S100P has been shown to impact upon the functionality of several key cancer hallmarks in the RL95-2 cell line and highlights the need to further investigate S100P in the context of endometrial cancer.
Supervisor: Hapangama, Dharani ; Barraclough, B. R. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral