The unconventional Myosin-1c (MYO1C) is located in a region of recurrent losses distal to tumor protein 53 (TP53) and has been proposed as a prominent tumour suppressor gene. Lowered expression of MYO1C is reported in endometrial cancer (EC). Earlier studies suggested a negative correlation between MYO1C protein level and activation of P13K/AKT signalling and cell proliferation. PI3K/AKT signaling pathway is known to play a critical role in cell proliferation and survival by regulation of the Forkhead Box O (FOXO) transcription factors. FOXOs have a critical role in cellular events such as cell differentiation, proliferation, survival, apoptosis and angiogenesis. FOXO proteins may have tumour suppressor characteristics since the loss of FOXO activities through hyperphosphorylation and/or lowered expression disable the ability of the cell to arrest at key cell cycle checkpoints and induction of apoptosis. Hence, FOXO proteins are suggested to be involved in the control of tumour growth through engaging with pro-apoptotic pathways as well as by blocking cell cycle progression.
This study was designed to analyze the expression levels of both active and inactive (phosphorylated forms) of FOXO proteins as correlated to MYO1C expression in a panel of clinical endometrial cancer samples. The overall objectives of the project were to set up Western blot protocols for the analysis of FOXO protein expression in EC samples, to investigate whether there is a correlation between levels of FOXO proteins and EC stages, and finally to examine for the potential correlation between inactive form of FOXO and MYO1C expression in these samples. Due to technical problems, in terms of protein expression correlation studies the results generated in this study were deemed inconclusive and no significant conclusions can be made. However, several troubleshooting approaches for Western blot analysis of FOXO proteins were practiced, which are presented in this paper that shall be useful for further research in the field.