AGEs通过JAK2/STAT3信号通路诱导结肠癌SW480细胞增殖、侵袭及上皮间质转化

Advanced glycation end products enhance proliferation and invasion of human colon cancer SW480 cells through JAK2/STAT3 pathway

目的观察糖基化终末产物(advanced glycation end products,AGEs)对结肠癌SW480细胞增殖、凋亡、侵袭、迁移及上皮间质转化的影响,并探讨其相关机制。方法将结肠癌SW480细胞分为对照组、AGEs组(200μg/m L AGEs)、AGEs(200μg/m L)+AG490(50μmol/L)组,CCK-8检测细胞增殖活力,流式细胞术检测细胞周期、凋亡,Transwell实验检测细胞体外侵袭能力,划痕实验检测细胞体外迁移能力。不同浓度(0、50、100、200μg/m L)AGEs和200μg/m L AGEs、50μmol/L AG490单独处理及二者联合处理SW480细胞后,Western blot检测上皮间质转化相关分子标志物E-cadherin、N-cadherin、Vimentin及JAK2/STAT3信号通路相关分子的变化。结果与对照组比较,AGEs能明显促进结肠癌SW480细胞的增殖,减少G1/S期阻滞,抑制凋亡,增强体外的侵袭、迁移能力(P<0.05)。与AGEs组比较,AGEs+AG490组结肠癌SW480细胞增殖减少,G1/S期阻滞增加,凋亡增加,侵袭、迁移能力降低,差异有统计学意义(P<0.05)。Western blot检测结果显示,AGEs处理结肠癌SW480细胞后E-cadherin表达降低,N-cadherin、Vimentin、p-STAT3、p-JAK2表达升高,而AG490可逆转AGEs对结肠癌SW480细胞的作用。结论 AGEs能够通过JAK2/STAT3信号通路促进结肠癌SW480细胞的增殖,抑制凋亡,增强侵袭、迁移能力,促进上皮间质转化。

Objective To explore the effects and the possible mechanisms of advanced glycation products （AGEs） on the proliferation,invasion and epithelial-mesenchymal transition（EMT） of human colon cancer cell line SW480. Methods The SW480 cells were divided into control group, AGEs （200 μg/mL） group, and AGEs （200 μg/mL）＋AG490（50 μmol/L） group. After the treatment, the cell proliferation, cell cycle and apoptosis, invasion and migration were tested by CCK-8 assay, flow cytometry, Transwell chamber test and wound healing assay. After the cells were treated with 0, 50, 100 and 200 μg/mL AGEs or 200 μg/mL AGEs plus 50 μmol/L AG490, the expression levels of EMT markers, E-cadherin, N-cadherin and Vimentin, and JAK2/STAT3 pathway molecules, STAT3, p-STAT3, JAK2 and p-JAK2 were detected by Western blotting. Results Compared with the control group, AGEs significantly promoted the proliferation,invasion and migration of SW480 cells, reduced the number of cells arrested at G1/S phase, and inhibited cell apoptosis （P〈0.05）. Whereas, these effects were obviously reversed by the addition of 50 μmol/L AG490 （P〈0.05）. Western blotting showed that AGEs increased the expression levels of N-cadherin, Vimentin, p-STAT3 and p-JAK2, and decreased that of E-cadherin. But AG490 reversed such changes resulted from AGEs treatment. Conclusion AGEs enhance proliferation, invasion and migration, and promote EMT in human colon cancer cell line SW480 through JAK2/STAT3 signaling pathway.