NCOA5 Is Highly Expressed In Colorectal Cancer And Modulates Cancer Cell Proliferation,migration And Invasion Via The PI3K/AKT Pathway

Objective: To detect the expression of NCOA5 in colorectal cancer and to analyze the relationship between the expression of NCOA5 and clinicopathological features of patients; To investigate the potential mechanisms on the cell proliferation, migration and invasion in different CRC cell lines.Methods: NCOA5 status was determined by performing immunohistochemistry(IHC) in 70 CRC patients’ tissues, and twelve paired tumor and adjacent normal specimens were examined by Western blot analysis. At the same time, five CRC cell lines including HT29, CCL244, HCE8693, SW620, SW480 were subjected to western blot for NCOA5 protein, respectively. Stable cell lines SW480 which were overexpressed of NCOA5 and SW620 which were knockdown of NCOA5 were established. Furthermore, knockdown of NCOA5 resulted in the suppression of proliferation, migration and invasion of SW620 cells as detected by CCK8 assay, cell-cycle analysis, wound-healing and transwell assay. Meanwhile, expression of p-AKT, AKT, Cyclin D1, P27, MMP9 was evaluated by western blot in SW480 and SW620 cells. PI3 K inhibitor LY294002 was used to further confirm the PI3K/AKT pathway.Results: The expression of NCOA5 in CRC Tissue samples was notably higher than that in the adjacent normal specimens. And it was found to be signifcantly correlated with length of tumor(P<0.001), regional Lymph nodes(P =0.005) and stage(P =0.018). Highly malignant cell lines, for example SW620, showed a high expession of NCOA5 than others. Knockdown of NCOA5 reduced the number of proliferated, migrated and invaded cells in SW620 cells. Furthermore, NCOA5 knockdown reduced AKT phosphorylation and suppressed Cyclin D1 and MMP9 activities, meanwhile it induced P27 activation in SW620 cells. The opposite result was gained in SW480 cells which were overexpressed of NCOA5. LY294002 could block the effect of PI3K/AKT/Cyclin D1, P27, MMP9 pathway in CRC cell lines.Conclusions: In the present study, we first demonstrated the high expression of NCOA5 in CRC patients’ tissues and its clinical implications. We further investigated role of NCOA5 which might contribute toumor cell proliferation, migration and invasion through activating PI3K/Akt signaling and subsequently increasing Cyclin D1 and MMP9,and also decreasing P27 in CRC cells.