IL-17A通过NF-κB通路介导MMP-2/9表达促进结肠癌SW480细胞迁移和侵袭

IL-17A promotes migration and invasion of colon cancer SW480 cells through NF-κB pathway mediating MMP2/9 expression

目的:探讨IL-17A对结肠癌细胞株SW480侵袭、迁移的作用及其机制。方法:体外培养结肠癌SW480细胞,分为实验组(IL-17A 50 ng/ml)及对照组(空白组)。通过细胞划痕实验观察细胞的迁移能力,Transwell侵袭实验检测细胞的侵袭能力,Western blotting检测细胞MMP-2/9蛋白及PI3k/AKT/NF-κB通路相关蛋白的表达水平。结果:经50 ng/ml的IL-17A处理后,(1)SW480细胞的迁移距离及穿膜细胞数目均明显增加[(2.49±0.18)vs(1.54±0.21)mm及(262.00±24.60)vs(92.00±31.16)个,均P<0.05];(2)SW480细胞MMP-2/9蛋白水平明显上调[(0.41±0.05)vs(0.23±0.03)及(0.76±0.09)vs(0.25±0.04),均P<0.05];(3)SW480细胞AKT磷酸化水平表达增加[(0.72±0.1)vs(0.28±0.04),P<0.05],P65和P50蛋白表达水平明显升高[(0.78±0.10)vs(0.35±0.04)和(0.85±0.15)vs(0.44±0.06),均P<0.05],而c-Rel、Re LB和P52蛋白表达无明显变化(P>0.05)。结论:IL-17A诱导结肠癌SW480细胞迁移和侵袭,其机制可能与激活PI3K/AKT/NF-κB转导通路、调节MMP-2/9蛋白表达有关。

Objective：To explore the effect and mechanism of IL-17A on invasion and migration of colon cancer SW480 cells.Methods： Colon cancer SW480 cells were in vitro cultured and divided into experimental group （IL-17A 50 ng/ml） and control group （blank group）. The migration ability was observed by cell scratch assay, the invasion feature of SW480 cells was detected by Transwell assay, and the expressions of MMP-2/9 protein and PI3k/AKT/NF-κ B pathway related proteins were tested by Western blotting. Results： After the treatment with IL-17A（50 ng/ml）, （1） the migration distance and trans-membrane number of SW480 cells significantly increased （[2.49±018] mm vs [1.54±0.21] mm; [262.00±24.60] vs [92.00±31.16]; all P〈0.05）; （2） the expression of MMP-2/9 protein in SW480 cells was obviously up-regulated （[0.41±0.05] vs [0.23±0.03]; [0.76±0.09] vs [0.25±0.04], all P〈0.05）; （3） AKT-phosphorylation level in SW480 cells was significantly increased （[0.72±0.1] vs [028±0.04], P〈0.05）, and the protein expressions of P65 and P50 were significantly increased （[0.78±0.10] vs [0.35±0.04]; [0.85±0.15] vs [0.44±0.06], all P〈0.05）; however, the protein expressions of c-Rel, ReLB, and P52 showed no significant difference （P〉0.05）. Conclusion： IL-17A induced the migration and invasion of colon cancer SW480 cells, and increased tumor cell viability; the mechanism may be related to the activation of PI3K/AKT/NF-κB pathway and up-regulation of MMP-2/9 protein expression.