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The role of prostate cancer microvesicles in epithelial mesenchymal transition

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Alokozai, Sarah (2019) The role of prostate cancer microvesicles in epithelial mesenchymal transition. Doctoral thesis, London Metropolitan University.

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Abstract

Microvesicles (MVs) are shed to the extracellular environment upon activation by both normal and diseased cells. Increasing evidence has shown that cancer cellderived MVs carry pathogenic components, such as proteins, messenger RNAs, microRNAs, long non-coding RNAs, DNAs, lipids, and transcriptional factors, which can mediate paracrine signaling in the tumor microenvironment, and that they are thus highly capable of altering the function of the recipient cells. MVs released from tumour cells are thought to play an important role in cancer metastasis. Most carcinomas are derived from epithelial cell lines, which normally adhere together tightly to form the structural foundations of many organs. Epithelial Mesenchymal Transition (EMT) has been linked to malignancy in many carcinomas of epithelial origin. In this study, I look at the role of PC3 (Prostate Cancer cell line) derived MVs in the induction of EMT in PNT2 (normal prostate) cells. Uptake of PKH26-labelled PC3-derived MVs by recipient cells was greater at a low pH of 5.4 and was dependent on MV-based protein and phosphatidylserine interaction with recipient cells.

Biochemical and morphological changes, indicative of EMT, (elongation, increased expression of mesenchymal marker Vimentin and reduced expression of epithelial marker, E-cadherin) were observed using fluorescent microscopy and flow cytometry. To further investigate the protein cargo of PC3, PNT2 and the transformed PNT2 (tPNT2) cells and their respective MVs LC-MS/MS analysis was carried out. Around 1290 proteins were identified collectively in all the samples.

Pathway analysis tools such as DAVID, PANTHER, KEGG and STRING were used to analyse the protein data. Importantly this study identified key proteins in PC3 MVs that may have contributed to the process of EMT, such as integrins which play a vital role in the biology of invasive carcinoma and setting up a pre-metastatic niche.

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