AGE/RAGE regulates the epithelial-mesenchymal transition and stemness maintenance of colorectal cancer cells / 肿瘤

ABSTRACT

Objective: To explore the effects of advanced glycation end products (AGEs) on the epithelialmesenchymal transition (EMT) and stemness maintenance of colorectal cancer HCT116 cells and its possible mechanism. Methods: The abilities of proliferation, migration, invasion, clone formation and sphere formation of HCT116 cells after treatment with 200 μg/mL AGEs and 200 μg/mL AGEs in combination with 50 μmol/L phosphatidylinositol-3 kinase (PI3K)-specific inhibitor LY294002 were detected by CCK-8, wound-healing assay, Transwell chamber assay, clone formation assay and sphere formation assay, respectively. After treatment with 200 μg/ mL AGEs and 200 μg/mL AGEs in combination with 50 μmol/L LY294002, respectively, the ability of invasion and the proportion of CD133plus signsphered HCT116 cells as well as the expressions of CD133, matrix metalloproteinase-2 (MMP-2) and MMP-9 in sphered HCT116 cells were detected by Transwell chamber assay, FCM and Western blotting, respectively. The expressions of the receptor of advanced glycation end products (RAGE), E-cadherin, N-cadherin, vimentin, Snail, PI3K, phosphorylated PI3K (p-PI3K), protein kinase B1 (PKB1, Akt1) and phosphorylated Akt1 (p-Akt1) in sphered HCT116 cells after treatment with different concentrations of AGEs, 200 μg/mL AGEs and 200 μg/mL AGEs in combination with 50 μmol/L LY294002 respectively were detected by Western blotting. Results: The abilities of migration, invasion, clone formation and sphere formation of HCT116 cells after treatment with 200 μg/mL AGEs were improved (P < 0.01, P < 0.05, P < 0.01, P < 0.01, P < 0.05), and these effects were inhibited by 50 μmol/L LY294002 (P < 0.05, P < 0.05, P < 0.01, P < 0.01, P < 0.05). After treatment with AGEs, the ability of invasion and the proportion of CD133plus sign sphered HCT116 cells were increased (P < 0.01, P < 0.05), and the expression levels of CD133, MMP-2 and MMP-9 were up-regulated (all P < 0.05), whereas these effects were opposite by 50 μmol/L LY294002 (P < 0.01, P < 0.05, all P < 0.05). The expression level of RAGE in HCT116 cells after treatment with different concentrations of AGEs was up-regulated (P < 0.05). After treatment with 200 μg/mL AGEs, the expression levels of p-PI3K, p-Akt1, N-cadherin, vimentin and Snail proteins were up-regulated (all P < 0.05), whereas the expression level of E-cadherin protein was down-regulated (P < 0.05); but these effects were opposite by 50 μmol/L LY294002 (all P < 0.05). Conclusion: AGE activates the RAGE signal pathway and improves the abilities of proliferation, migration, invasion, clone formation and stemness maintenance of colorectal cancer HCT116 cells, and this mechanism may be related to EMT mediated by AGE/RAGE/PI3K-Akt signal pathway.