The IGF system is a crucial regulator of normal growth and development, however dysregulation of the system on multiple levels is associated with the incidence of a wide variety of malignancies including the breast, thyroid, lung, and colon, making the IGF system an important anti-cancer therapeutic target. Due to its role in mediating cellular proliferation, protection from apoptosis, and metastasis, traditional focus has been set on examining the role of the type 1 IGF receptor [IGF1R] in cancer. However there is mounting evidence to suggest the insulin receptor [IR] may also be involved in the potentiation and pathogenesis of cancers. The observation that IGF-II is overexpressed, compared to normal tissues, by cancers suggests signaling via target receptors by this ligand has important implications on cancer pathogenesis. Indeed, both the IGF1R and IR have been demonstrated to be up-regulated in a variety of malignancies. In regards to IR isoform, the IGF-II binding IR-A is preferentially expressed by a number of cancer cell types. Together with the observation that an autocrine proliferative loop exists between IGF-II and the IR-A in malignant thyrocytes and cultured breast cancer cells, suggests signaling via the IR-A may play a role in cancer cell growth and survival. However, very few studies on the IR-A have been conducted in cells co-expressing the IGF1R. This is mainly due to the difficulties associated with discrimination between signaling arising from IGF1R homodimers, IR-A homodimers, and IGF1R/IR-A hybrid receptors. It is not known how the IR-A interacts, and functions in conjunction with the other receptors of the IGF system to signal biologically relevant outcomes, especially in terms of anti-cancer therapeutics that aim to block and down-regulate the IGF1R. Current anti-cancer therapies targeting the IGF system have concentrated on blocking IGF signaling via the IGF1R, due mostly to the functional properties of the receptor, but also in part due to the metabolic consequences associated with blockade and inhibition of the IR. This individual targeting of the IGF1R potentially leaves a pathway by which IGF-II secreted by the tumour can circumvent current IGF1R based therapies. Consequently, this thesis investigated whether the IR-A could compensate for the targeted loss of the IGF1R and how the IR-A interacts with the IGF1R in cells co-expressing these two receptors. In addition, the individual ability of the IR isoforms to signal biological outcomes in response to IGF stimulation was assessed. The main experimental techniques used throughout this body of work included; assessment of protein expression and activation by Western blot, siRNA mediated gene silencing, and measures of cell proliferation, survival, and migration. The key areas of investigation included: 1. Investigation of the individual ability of the IR isoforms to signal biological outcomes in response to IGF stimulation 2. Identification of an appropriate cell line model in which to investigate the interactions between the IR-A and IGF1R 3. Optimisation of siRNA mediated knock-down of the IR-A and IGF1R in SW480 colorectal adenocarcinoma cells 4. Determination of the biological role of the IR-A in SW480 cells co-expressing the IGF1R The key findings from this work included: 1. The IR-A could not compensate for IGF1R depletion in SW480 cells 2. Dual silencing of the IR-A and IGF1R indicated signaling via the IGF1R was dominant to signaling via the IR-A in SW480 cells 3. Signaling via IR-A/IGF1R hybrid receptors may not be as potent as signaling via IGF1R homodimers 4. IGF-I at physiological concentrations can stimulate biological responses via both isoforms of the IR. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1337339 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 2008
Identifer | oai:union.ndltd.org:ADTP/272796 |
Date | January 2008 |
Creators | Brierley, Gemma Victoria |
Source Sets | Australiasian Digital Theses Program |
Detected Language | English |
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