ROS/p38MAPK通路对结直肠癌SW480细胞侵袭能力的影响

Effect of ROS/p38MAPK Pathway on Invasion of Colorectal Cancer SW480 Cells

目的探究ROS/p38MAPK通路对PKC诱导人结直肠癌SW480细胞MMP-9表达及细胞侵袭的作用机制。方法利用蛋白激酶C(PKC)激动剂12-O-十四烷基酚-13-乙酸酯(TPA)处理人结直肠癌SW480细胞,并且进行PKC抑制剂/ROS抑制剂/p38抑制剂干扰。通过Western blot法检测MMP-9、p38/p-p38以及核内p65蛋白表达水平;激光共聚焦检测细胞内ROS含量;明胶酶谱法检测MMP-9细胞外分泌情况,Transwell实验检测细胞侵袭能力。结果TPA增加细胞内ROS的生成和p38的磷酸化,可诱导p65和MMP-9的表达上调;而预处理PKC抑制剂/ROS抑制剂/p38抑制剂能够阻断TPA诱导的ROS生成和p38磷酸化,下调核内p65蛋白的表达,进而使TPA诱导的SW480细胞MMP-9表达及细胞侵袭能力显著下降。结论ROS介导p38MAPK信号通路调控转录因子NF-κB,促进结直肠癌细胞MMP-9表达及侵袭发生;PKC诱导SW480细胞侵袭中的潜在作用机制,为结直肠癌转移研究提供新的思路。

Objective To investigate the mechanism of ROS/p38MAPK pathway on MMP-9 expression and cell invasion in human colorectal cancer SW480 cells induced by PKC.Methods Human colorectal cancer SW480 cells was treated with protein kinase C(PKC)agonist 12-O-tetradecylphenol-13-acetate(TPA),and PKC inhibitor/ROS inhibitor/p38 inhibitor interference was performed.Western blot was used to detect the expression of MMP-9,p38/p-p38,and nuclear p65 protein.The intracellular reactive oxygen species(ROS)content was detected by confocal microscope and the secretion of MMP-9 was detected by gelatin zymography assay,and the invasion ability of cells was detected by Transwell test.Results TPA increases the generation of ROS and phosphorylation of p38 in cells,which can induce the up-regulation of the expression of p65 and MMP-9.The pretreatment of PKC inhibitor/ROS inhibitor/p38 inhibitor can block TPA-induced ROS generation and p38 phosphorylation,and down-regulation of the expression of p65 protein in the nucleus.Then TPA-induced expression of MMP-9 and cell invasion ability were significantly reduced in SW480 cells.Conclusion ROS mediates p38MAPK signaling pathways to regulate transcription factor NF-κB for further promoting MMP-9 expression and invasion of colorectal cancer cells.It indicates the potential mechanism of PKC-induced SW480 cell invasion and provides new ideas for the research on colorectal cancer metastasis.