Introduction: Colorectal cancer is caused by the abnormal division of cells within the epithelial lining of the colon. Growing body of evidence suggests that tumour initiation and progression is caused by a small subset of cells called cancer stem cells.
Studies have suggested that two novel genes, GAEC1 and JK-1, are involved in the initiation and early establishment of tumours through their involvement in cell proliferation and formation of tumours in nude mice.
Objectives: The project aims to identify novel colorecal cancer stem cell genes involved in tumour formation.
Methodology: Human colorectal cancer stem cells were isolated from the remaining populations of two colorectal cancer cell lines, SW480 and SW48. The stem cells were first isolated based on their expression of the stem cell marker ALDH1 using the Aldeflour Assay kit, before being sorted from the remaining population based on their expression of a second stem cell marker, CD44, using flow cytometry. The cells were then subjected to a self-renewal sphere formation assay. The stem-like phenotype was confirmed using qPCR and Western blot methodology to observe their expression of other stem cell markers such as SOX-2, Notch, Oct-3/4, Sail1 and Twist. The expression of the novel cancer genes GAEC1 and JK-1 was also determined using the qPCR and Western blot methodology. Expression of these markers was normalised to the SW480 and SW48 cell population.
Results: Cancer stem-like cells were isolated from the remaining population. Expression of the stem cell markers and novel genes GAEC1 and JK-1 varied between the isolated stem-like cell population and the control SW480 and SW48 cells.
Conclusion: Our primary findings indicate the importance of GAEC1 and JK1 in colorectal cancer initiation. Further functional studies will need to be done to further unravel the role that these genes play in colorectal cancer