目的 探讨欧前胡素对人胃癌细胞(MKN-45)和人结肠癌细胞(HT-29)中烟酰胺腺嘌呤二核苷酸磷酸氧化酶(Noxs)-活性氧(ROS)氧化应激通路的作用及对肿瘤细胞增殖活性的影响。方法 MKN-45和HT-29细胞经过不同浓度(3、10、30、100 μmol/L)的欧前胡素及阳性对照二苯基氯化碘盐(DPI)处理后,使用CCK8法检测细胞增殖活性,使用DHE荧光探针检测细胞内ROS水平,使用Western blotting检测细胞内Nox1、Nox2、Nox3、Nox4、Nox5家族蛋白表达。结果 CCK8法结果显示,欧前胡素可浓度相关性地抑制MKN-45和HT-29细胞增殖活性;DHE荧光探针法结果显示,欧前胡素干预可使MKN-45和HT-29细胞荧光强度明显减弱,ROS水平降低;Western blotting结果显示,给予欧前胡素可使MKN-45细胞Nox1、Nox2、Nox3、Nox4、Nox5蛋白表达水平显著降低(P<0.05、0.01、0.001),使HT-29细胞Nox1、Nox2蛋白表达水平显著降低(P<0.05、0.01)。结论 欧前胡素可通过抑制MKN-45和HT-29细胞中Noxs-ROS通路,诱导细胞增殖活性下降,发挥抗肿瘤作用。

Objective To explore the effect of imperatorin on the Noxs-ROS oxidative stress pathway and proliferative activity in human gastric cancer cells (MKN-45) and human colon cancer cells cells (HT-29). Methods MKN-45 and HT-29 cells were administered with different concentrations (3, 10, 30, and 100 μmol/L) of imperatorin and positive control diphenyl iodide salt chloride (DPI), CCK8 method was used to detectcell proliferation activity, intracellular ROS level was detected by DHE fluorescent probe, and Western blotting was used to detect the protein expressions of Nox1, Nox2, Nox3, Nox4, and Nox5. Results CCK8 assay showed that imperatorin could concentration-dependently inhibited the proliferation activity of MKN-45 and HT-29 cells. The results of DHE fluorescent probe assay showed that imperatorin intervention could significantly reduced the fluorescence intensity and ROS level of MKN-45 and HT-29 cells. Western blotting results showed that administration of imperatorin could reduced the protein expression level of Nox1, Nox2, Nox3, Nox4, and Nox5 in MKN-45, and Nox1 and Nox2 in HT-29 cells. Conclusion Imperatorincan inhibits the proliferative activity of MKN-45 and HT-29 cells by down-regulating of Noxs-ROS pathway, and exhibits anti-tumor effects.

R285.5

国家自然科学基金资助面上项目(82172418)