Our Methods
Along with standard cell culturing, molecular biology techniques (Western blotting, qRT-PCR), proliferation assays, and immunohistochemical methods, our research group uses a range of innovative models to drive forward our research goals.
Stably tranfected cells
We have significant experience in creating novel stable transfected cancer cell lines that more closely mimic the clinical condition. Successful development of these cells lines have aided our understanding
of the role of steroid sulphatase in breast cancer (see here and here), 17beta-hydroxysteroid dehydrogenase type-1 in breast cancer (see here), and 17beta-hydroxysteroid dehydrogenase type-3 in prostate cancer (see here and here).
We are currently developing novel colorectal cancer cell lines that over
express key oestrogenic enzymes in orrder to further understand the role oestrogens play in colorectal cancer progression.
Novel LC/MS methods
Using expertise within the Centre for Endocrinology, Diabetes, and Metabolism at the University of Birmingham, we use highly sensetive LC/MS technology to accurately measure various steroids in cell media,
mouse serum, and human tissue samples.
In collaboration with the MUSCLE Project (see here), we have recently developed a novel LC/MS methodology that will sensitively quantify oestrone, oestradiol, and conjugated oestrogens in a single sample at low cocentrations. These innovative techniques will allow us a greater understanding of oestrogen metabolism pathways in various cancers.
Steroid Enzyme Assays
In vivo Cancer Models
To examine the steroid metabolising capabilites of human and animal tissue, annd cancer cell lines, we use various radio-labelled enzyme
assays to measure steroid sulphatase and aromatase activity. We also have experience in using TLC to determine the activity of various other steroidogenic enzyme activity.
These assays have been vital for our ability to measure efficacy of steroid sulphatase and aromatase inhibitors in a range of cancer models (see here, here, and here).
We have over a decades experience in developing novel, innovative in vivo cancer models. Many of these have been used for the successful pre-clinical
development of various novel anti-cancer compounds. For example, our work was intrumental in providing data to support the further clinical development of the breast cancer drug Irusostat (see here and here).
Using this expertise, we are currently developing novel colorectal cancer and adrenocortical carcinoma mouse models.
Mitochondrial bioenergetics
Previously, we have used various techniques to measure mitochondrial bioenergetics. In particular, our group has utilised the proteo-metric dye
JC-1 to directly monitor the electron transport chain and its response to various pharmacological interventions, particularly anti-cancer agents.
We are now using these techniques to examine the role of NNT in adrenocortical carcinoma and whether this mitochondrial protein is a target for this particular cancer.
Flow Cytometry
We have previously developed novel flow cytometry methods that measure apoptosis and cell cycle in ex vivo tumour samples (see here). These methods have allowed for the direct measurement of these parameters in treated tumours, allow for direct assessment of treatment efficacy.
Consequently, our group has signficant experience in both novel and standard flow cytometric techniques. This allows us to measure proliferation, apoptosis, and cell cycle.