- Title
- The role of cancer procoagulant on the MTOR pathway
- Creator
- Chiuswa, Chengetanai
- Subject
- Cancer -- Research
- Subject
- Neovascularization
- Subject
- Biochemistry
- Subject
- Blood coagulation factors -- Biotechnology
- Date Issued
- 2016
- Date
- 2016
- Type
- Thesis
- Type
- Masters
- Type
- MSc
- Identifier
- http://hdl.handle.net/10948/7303
- Identifier
- vital:21316
- Description
- Cancer procoagulant (CP) is a cysteine protease found in tumour cells and amnion chorion membranes. The main function of CP is not yet known, but it has potential roles in tumour growth and metastasis. Initially, CP was believed to increase coagulation in cancer patients; however, research has shown that increase in CP concentration does not correlate with an increase in coagulation. The location of CP in amnion chorion membranes and tumour cells only led to the hypothesis that CP might be involved in inducing blood vessel and or lymph vessel formation. CP was shown to induce lymphangiogenesis (lymph vessel formation) in human telomerase reverse transcriptase-human dermal endothelial cells (hTERT-HDLEC) (Tshaka, 2011). CP-induced tube formation was inhibited by rapamycin; indicating that CP may be signalling via the mammalian target of rapamycin (mTOR) pathway. The aim of this study was to investigate the effect of CP on the mTOR signalling pathway using human umbilical endothelial vein cells (HUVECs) as a model. CP was isolated from amnion chorion membranes and purified using two anion exchange chromatography steps. Purified CP (2 μg/ml) was used to induce tube formation in endothelial cells (HUVECs) seeded on growth factor reduced (GFR) Matrigel. In addition, the 2 μg/ml CP was used to treat cultured HUVECs. Sodium dodecyl sulphate polyacrylamide electrophoresis (SDS-PAGE) and western blotting were used to determine phosphorylation levels of protein kinase B (Akt) and ribosomal protein S6 kinase (S6K). CP was successfully isolated and purified using anion exchange chromatography. The effect of CP on tube formation was not significant relative to the control in the HUVEC cell line. The role of CP on Akt and S6K phosphorylation still needs to be verified by using sensitive methods of quantification such as enhanced chemiluminescence (ECL) and enzyme linked immunosorbent assay (ELISA). The levels of CP activity were shown to be higher in early tumour growth than in advanced cancer suggesting that certain physiological factors could be increasing CP activity during early tumour growth. This study investigated the effect of cobalt chloride on CP activity in breast cancer cell lines. Cobalt chloride reduced CP activity in MCF-7 and promoted CP activity in MDA-MB-231.
- Format
- xiii, 101 leaves
- Format
- Publisher
- Nelson Mandela Metropolitan University
- Publisher
- Faculty of Science
- Language
- English
- Rights
- Nelson Mandela Metropolitan University
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