The insulin-like growth factor system and adenocarcinoma of the colon

The insulin-like growth factor (IGF) system is important in normal growth and development. However, it is also known to be involved with malignant transformation and cellular proliferation. IGF binding proteins modulate the biological activity of IGF-I, either potentiating or inhibiting its activity, as well as determining how much enters the circulation at any one time. IGF binding protein-4 (IGFBP-4), for example is believed to be inhibitory to the effects of IGF-I. This thesis shows that the colon cancer cell lines Colo 205, HT29 and WiDR proliferate in response to IGF-I, and that IGFBP-4 at high concentrations inhibits their growth. However, it was found that with lower concentrationsof IGFBP-4, proliferation in HT29 and WiDR cells increased. Nevertheless in two cell lines, IGFBP-4 partially negated the proliferative effects of IGF-I. An antibody against IGFBP-4 was used to show that endogenous IGFBP-4 plays an important role in modifying cell growth. In order to start in vivo experiments which required considerable quantities of IGFBP-4, this protein was produced in an expression system and purified using an immunoaffinity column method. The rhIGFBP-4 thus produced was shown to be functional and to inhibit colorectal cancer cell growth in vitro. A nude mouse model of colon cancer was produced and the expression of components of the IGF system in this model determined using PCR. Experiments were performed using conditioned medium from Colo 205 cells to investigate IGFBP-4 protease activity. This thesis shows that manipulation of the IGF system is a potential target for further research into treatment for adenocarcinoma of the colon.