槐耳浸膏通过调控ROS抑制结直肠癌细胞MMP-9表达及侵袭能力的研究

Experimental study of Huaier extract inhibiting MMP-9 expression and invasion ability of colorectal cancer cells by regulating ROS

目的:探讨槐耳浸膏对结直肠癌细胞活性氧(ROS)的调控作用及细胞侵袭能力的影响。方法:利用PKC激动剂佛波酯(TPA)处理人结直肠癌SW480细胞,倒置显微镜下观察TPA诱导细胞形态学的改变;MTT法检测不同浓度槐耳浸膏对细胞增殖的影响;Western blot检测基质金属蛋白酶-9(MMP-9)和核内p50、p65蛋白表达水平;明胶酶谱法检测MMP-9细胞外分泌情况;流式细胞仪测定细胞内ROS含量;Transwell侵袭实验检测细胞的侵袭能力。结果:TPA诱导下SW480细胞由上皮样转变为间叶样细胞形态,可诱导MMP-9蛋白的表达上调。槐耳浸膏浓度依赖性抑制SW480细胞的增殖;可阻断TPA诱导的ROS生成,下调核内p50和p65蛋白表达;抑制MMP-9蛋白表达和细胞外MMP-9分泌能力,且可抑制TPA诱导的细胞侵袭能力。结论:槐耳浸膏通过调控ROS/NF-κB信号通路抑制MMP-9的表达,阻断了TPA诱导的结直肠癌SW480细胞的侵袭。

Objective: To explore the effect of Huaier extract on the regulation of colorectal cancer cell reactive oxygen species(ROS) and cell invasion ability. Methods: The PKC agonist phorbol ester(TPA) was used to treat human colorectal cancer SW480 cells, and the morphological changes induced by TPA were observed under an inverted microscope.The MTT method was used to detect the effect of different concentrations of Huaier extract on cell proliferation. Western blot detects the expression levels of MMP-9 and p50/p65 proteins in the nucleus. Gelatin zymography detects the extracellular secretion of MMP-9.Flow cytometry determines the intracellular ROS content. Transwell invasion test detects the invasion ability of cells. Results: TPA-induced SW480 cells change from epithelial-like to mesenchymal-like cell morphology, which can induce the up-regulation of MMP-9 protein expression;Huaier extract concentration-dependently inhibits the proliferation of SW480 cells;it can block the generation of ROS induced by TPA and down-regulate The expression of p50 and p65 proteins in the nucleus;inhibits the expression of MMP-9 protein and the ability of extracellular MMP-9 secretion, and can inhibit the cell invasion ability induced by TPA. Conclusion: Huaier extract inhibits the expression of MMP-9 by regulating the ROS/NF-κB signaling pathway, blocking the invasion of colorectal cancer SW480 cells induced by TPA.