Investigation of potential CTCF regulatory targets in breast cancer cells
CTCF is a multifunctional transcription factor and a candidate tumour suppressor gene. Surprisingly for a tumour suppressor the levels of CTCF in breast cancer cell lines and tumours were found to be elevated compared to breast cell lines with finite life span and normal breast tissues. We proposed that up-regulation of CTCF may be linked to resistance of breast cancer cells to apoptosis.
For this purposed we used a proteomics approach to compare protein profiles in two different physiological situations in the breast cancer cell line ZR75-1 after transfection with CTCF siRNA and non-target control siRNA (control). Extracts were subjected to the two-dimensional gel electrophoresis (2D gel). Differentially expressed proteins were excised from the gels and analyzed by Mass Spectrometry: MALDI-TOF MS/MS or LC-MS/MS. A number of differentially expressed proteins were identified.
The aim of this project is to validate the expression of a panel of these proteins.
The role of CTCF and BORIS in prostate tumourigenesis
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CTCF is a new tumour suppressor gene and BORIS which is a paralogue of CTCF has features of an oncogene. The major objective of this proposal is to provide direct evidence that CTCF and BORIS are important in the pathogenesis of the prostate cancers.
Specifically we plan to:Establish the role of CTCF and BORIS in prostate tumourigenesis using a panel of prostate cell lines.
Investigate the connection between CTCF and BORIS gene abnormal structure and/or expression and prostate cancers.Rationale design approaches and measurements: The projects which are currently are carried out in our laboratory are focussed on biochemical and functional characterization of CTCF and BORIS. All the knowledge technology and expertise to perform various biochemical immunological and functional tests in vitro and in vivo in different cellular models for the thorough analysis of CTCF and BORIS have been accumulated in our laboratory. Generation of cellular models for constitutive expression of CTCF and BORIS.
Generation of cellular models for inducible expression of the CTCF and BORIS.
Generation of cellular models for transient CTCF and BORIS knock down. Functional characterization cells and ability of the wtCTCF to normalize the malignant phenotype could be of major importance for understanding prostate tumourigenesis. Future studies conceivably might lead to the development of molecular tools relevant to prostate cancer diagnosis prognosis and possibly lay ground for a therapeutic use of CTCF.
Validation of novel blood biomarkers for the monitoring of systemic therapy in metastatic breast cancer
The purpose of this investigation is to characterise identified potential biomarkers in the blood of patients with metastatic breast cancer undergoing chemotherapy or hormone therapy. Currently, the assessment of response to therapy relies on both subjective and objective measures. A series of tests ( Real-Time-qPCR immunohistochemistry Western blot assay and others) will be used to demonstrate the biomarker properties of the selected candidates. The discovery of a suitably sensitive blood biomarker could potentially replace and/or complement the assessment currently performed using imaging such as CT Isotope bone scans and MRI.
Understanding the Roles of CTCF and BORIS in the Hematopoietic System
CTCF and its paralogue BORIS share the same DNA-binding domain. While CTCF is ubiquitous and has features of a tumour suppressor BORIS is expressed in the testis but activated in various cancers (e.g. in