The metastatic spread of cancer cells to distant sites in the body accounts for the majority of cancer-related death and significantly decreases patient survival. Whilst cell migration is a physiologically important process, when uncontrolled, it can be a contributing factor to the metastatic phenotype. Actin polymerisation enables the dynamic restructuring of the cytoskeleton which is fundamental to cell migration and is stimulated by the Arp (actin-related protein) 2/3 protein complex which in turn is activated by members of the WAVE (WASP Verprolin homologous protein) family. WAVE1 and 3 expression was targeted separately in the PC-3 cell line utilising ribozyme transgene transfection. In vitro experiments revealed a reduction in cell growth and invasion following WAVE1 or 3 knockdown in PC-3 cells. These experiments were also repeated with small molecule inhibitors targeting the Arp2/3 complex, ROCK and N-WASP independently. This inhibitor work implicates Arp2/3 as a facilitator of cell proliferation through which WAVE regulates. Inhibition of Arp2/3, ROCK or N-WASP in WAVE1 knockdown cells increased cell invasion which may be attributed to the regulatory role of WAVE3 on MMP activity. Co-localisation of WAVE1 and 3 with ARP2 and ROCK-I was observed in PC-3 cells whilst this affect was abolished with WAVE1 or 3 knockdown. Furthermore, WAVE3 and WAVE1 knockdown affected ARP2 and ROCK-II tyrosine phosphorylation, respectively. These results suggest WAVE1 and 3 proteins are involved in several metastatic traits that characterise PC-3 cells. Furthermore, the contribution of WAVE in the networks that influence these traits may also involve association with Arp2/3 complex, ROCK-I and –II and N-WASP. Additionally, it sheds light on the similarities between these two related proteins and also highlights their subtle distinctions in PC-3 cells. The data outlined here provides justification to futher explore WAVE1 and 3 as potential contributors of prostate cancer progression.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:637137 |
| Date | January 2014 |
| Creators | Weeks, Hoi Ping |
| Publisher | Cardiff University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://orca.cf.ac.uk/70578/ |
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