Determination of the role of WSB-1 in breast cancer and metastasis to the bone

Lovell, Sapphire

Abstract

Breast cancer is the second most common cause of female-related deaths in the United Kingdom with metastatic spread the main reason for treatment failure and cancer-related deaths. The bone is the most frequent site of secondary cancer, significantly decreasing patient's quality of life and survival. WSB-1 is a hypoxic-responsive E3 ubiquitin ligase which has been found to be upregulated in metastatic cells. Furthermore, studies investigating WSB-1 have found it to be involved in the progression of various tumour types, including metastatic breast cancer, but specific roles in bone metastasis have not yet been identified. This study aims to identify potential pathways in breast cancer bone metastasis.
In order to do this, first in silico bioinformatic analysis of available RNA-sequencing data from the lab for WSB-1 knockdown was performed to identify significantly regulated bone metastasis factors potentially downstream of WSB-1. These were then validated through qPCR, ELISA and western blotting after siRNA treatment. Further identification of clinical relevance was then explored through in silico analysis of publicly available breast cancer patient datasets for correlation in gene expression between WSB1 and candidate genes.
Overall, in MDA-231-IV breast cancer bone homing cells, WSB-1 knockdown led to significant downregulation of ICAM1, IL1B, MMP1, MMP9, TGFBR1, PTGS2 and PTHLH, which are all key genes associated with breast cancer and bone metastasis. Further analysis of IL1 protein levels was explored using western blotting and ELISA, however, no significant differences was found when the knockdown of WSB-1 was present. Finally, in silico analysis further validated the correlation between WSB-1 and these factors in breast cancer patient mRNA expression datasets.
To conclude, WSB-1 could be involved in the biology of breast cancer bone metastasis by enhancing the vicious cycle of bone metastasis, but further validation of the identified factors and their impact in bone metastasis biology needs to be investigated. Further work could validate WSB-1 to be used clinically as a prognostic biomarker, with high levels indicating to drive metastatic potential or a predictive biomarker with the potential to inhibit WSB-1 using IL1 antibodies, and gene inhibitors such as ICAM1, MMP1, TGFBR1 and PTHLH. With further research, these treatment options could be successful and therefore aid in increasing distant metastasis-free survival and potentially patient overall survival rates.

Citation

Lovell, S. (2024). Determination of the role of WSB-1 in breast cancer and metastasis to the bone. (Thesis). University of Hull. https://hull-repository.worktribe.com/output/5086180

• Thesis Type: Thesis
• Deposit Date: Mar 20, 2025
• Publicly Available Date: Apr 11, 2025
• Keywords: Biomedical science
• Public URL: https://hull-repository.worktribe.com/output/5086180
• Additional Information: Faculty of Health Sciences; University of Hull
• Award Date: Dec 16, 2024