目的 探討歐前胡素對(duì)人胃癌細(xì)胞(MKN-45)和人結(jié)腸癌細(xì)胞(HT-29)中煙酰胺腺嘌呤二核苷酸磷酸氧化酶(Noxs)-活性氧(ROS)氧化應(yīng)激通路的作用及對(duì)腫瘤細(xì)胞增殖活性的影響。方法 MKN-45和HT-29細(xì)胞經(jīng)過(guò)不同濃度(3、10、30、100 μmol/L)的歐前胡素及陽(yáng)性對(duì)照二苯基氯化碘鹽(DPI)處理后,使用CCK8法檢測(cè)細(xì)胞增殖活性,使用DHE熒光探針檢測(cè)細(xì)胞內(nèi)ROS水平,使用Western blotting檢測(cè)細(xì)胞內(nèi)Nox1、Nox2、Nox3、Nox4、Nox5家族蛋白表達(dá)。結(jié)果 CCK8法結(jié)果顯示,歐前胡素可濃度相關(guān)性地抑制MKN-45和HT-29細(xì)胞增殖活性;DHE熒光探針?lè)ńY(jié)果顯示,歐前胡素干預(yù)可使MKN-45和HT-29細(xì)胞熒光強(qiáng)度明顯減弱,ROS水平降低;Western blotting結(jié)果顯示,給予歐前胡素可使MKN-45細(xì)胞Nox1、Nox2、Nox3、Nox4、Nox5蛋白表達(dá)水平顯著降低(P<0.05、0.01、0.001),使HT-29細(xì)胞Nox1、Nox2蛋白表達(dá)水平顯著降低(P<0.05、0.01)。結(jié)論 歐前胡素可通過(guò)抑制MKN-45和HT-29細(xì)胞中Noxs-ROS通路,誘導(dǎo)細(xì)胞增殖活性下降,發(fā)揮抗腫瘤作用。

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

國(guó)家自然科學(xué)基金資助面上項(xiàng)目(82172418)