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目的:探讨终末糖基化产物(advanced glycation end products,AGEs)对结肠癌SW620细胞上皮间质转化(epithelial-to-mesenchymal transition,EMT)及肿瘤干细胞标志物CD133的影响及其作用机制。方法:用不同浓度(0、50、100、200μg/ml)的AGEs处理SW620细胞后,采用划痕实验检测细胞的迁移能力;Transwell小室检测细胞的侵袭能力;流式细胞术检测CD133~+细胞的含量;Western blotting检测AGEs受体(receptor of AGEs,RAGE)、E-cadherin、Vimentin、ERK1/2、p-ERK1/2、CD133蛋白的表达情况。结果:与对照组(0μg/ml)比较,AGEs处理组(50、100、200μg/ml)在AGEs作用后,SW620细胞24h迁移距离[(1.55±0.15)、(1.58±0.19)、(1.75±0.21)vs(0.95±0.18)mm,均P<0.05]及48 h迁移距离[(2.11±0.22)、(2.21±0.37)、(2.68±0.23)vs(1.60±0.24)mm,均P<0.05]均明显增加;穿过Matrigel胶的数量明显增加[(176±19.52)、(194±17.70)、(220±25.5)vs(125±26.06)个,均P<0.05];CD133~+细胞比例明显增加[(4.75±1.49)、(10.34±1.54)、(14.45±2.41)%vs(0.77±0.41),均P<0.05]。与对照组比较,AGEs处理组(50、100、200μg/ml)Vimentin、RAGE、p-Erk1/2、CD133蛋白表达明显增加;而ERK1/2蛋白无明显变化;E-cadherin蛋白表达明显减少。结论:AGEs可以提高结肠癌SW620细胞体外的侵袭迁移能力,促进EMT的发生,诱导肿瘤干细胞的生成。其机制可能通过AGE-RAGE受体配体的激活,上调p-ERK1/2,从而调控EMT相关蛋白的表达,促进肿瘤干细胞的生成。
Objective: To investigate the effect of advanced glycation end products (AGEs) on the epithelial-to-mesenchymal transition (EMT) and the tumor stem cell marker CD133 in human colon cancer SW620 cells. Methods: SW620 cells were treated with different concentrations of AGEs (0, 50, 100, 200μg / ml), and the migration ability of SW620 cells was detected by scratch test. The invasion ability of SW620 cells was detected by Transwell chamber. Flow cytometry was used to detect CD133 ~ + cells . The expressions of AGEs receptor (RAGE), E-cadherin, Vimentin, ERK1 / 2, p-ERK1 / 2 and CD133 were detected by Western blotting. Results: Compared with the control group (0μg / ml), the migration of SW620 cells after exposure to AGEs for 50, 200 and 200μg / ml AGEs treatment group was significantly higher than that of the control group (1.55 ± 0.15, 1.58 ± 0.19 and 1.75 ± 0.21 ± 0.37), (2.68 ± 0.23) vs (1.60 ± 0.24) mm respectively (P <0.05) and the migration distance at 48 hours (P <0.05) ] (P <0.05). The number of CD133 ~ + cells increased significantly ([176 ± 19.52], (194 ± 17.70), (220 ± 25.5) vs (125 ± 26.06) (4.75 ± 1.49), (10.34 ± 1.54), (14.45 ± 2.41)% vs (0.77 ± 0.41), all P <0.05]. Compared with the control group, the expressions of Vimentin, RAGE, p-Erk1 / 2 and CD133 in AGEs treatment group (50, 100 and 200μg / ml) increased significantly, while there was no significant change in ERK1 / 2 protein expression and E-cadherin protein expression. CONCLUSION: AGEs can enhance the invasion and migration of colon cancer SW620 cells in vitro, promote the development of EMT and induce the generation of tumor stem cells. The mechanism may be through the activation of AGE-RAGE receptor ligand, upregulation of p-ERK1 / 2, thereby regulating the expression of EMT-related proteins, and promote cancer stem cell formation.