七甲氧基黄酮抑制人结直肠癌细胞增殖、迁移和侵袭的机制研究

Effect of heptamethoxyflavone on proliferation,migration and invasion of human colorectal cancer cells and its mechanism

目的:探究3,5,6,7,8,3',4'-七甲氧基黄酮(3,5,6,7,8,3',4'-heptamethoxyflavone,HMF)对人结直肠癌(colorectal cancer,CRC)细胞株SW480和HCT116增殖、侵袭和迁移的影响,并初步探讨其分子机制。方法:体外培养的SW480和HCT116细胞用不同浓度(0、12.5、25和50μmol/L)的HMF处理48 h后,采用CCK-8和集落形成实验检测细胞增殖能力;Transwell法检测细胞侵袭能力;划痕实验检测细胞迁移速率;通过DCF-DA活性氧实验分析其氧化应激水平;通过RT-qPCR检测血红素加氧酶1(heme oxygenase-1,HO-1)和NAD(P)H:醌氧化还原酶1[NAD(P)H:quinone oxidoreductase-1,NQO-1]的mRNA表达水平。结果:CCK-8和集落形成实验显示,SW480和HCT116细胞经过不同浓度的HMF处理后增殖能力显著减弱(P<0.05)。Transwell实验证明,HMF处理后SW480和HCT116细胞的侵袭能力显著减弱(P<0.05)。划痕实验结果表明,经过HMF处理,SW480和HCT116细胞的迁移能力受到明显的限制(P<0.05)。DCF-DA染色结果提示,HMF作用后SW480和HCT116细胞中的活性氧水平显著升高(P<0.05);进一步的RT-qPCR实验结果表明,HMF可显著抑制抗氧化分子HO-1和NQO-1的mRNA表达。结论:HMF可能通过诱导人CRC细胞发生氧化应激反应,进而抑制其增殖、侵袭和迁移。

AIM:This study is aimed to investigate the impact of 3,5,6,7,8,3',4'-heptamethoxyflavone(HMF)on the proliferation,invasion,and migration of human colorectal cancer(CRC)cell lines(SW480 and HCT116)and preliminarily explore the underlying molecular mechanisms.METHODS:Human colorectal cancer cells(SW480 and HCT116)cultured in vitro were subjected to various concentrations of HMF(0,12.5,25 and 50μmol/L)for 48 h.Proliferation levels were assessed using the CCK-8 assay,invasion abilities were examined via the Transwell assay,migration rates were measured using the scratch assay,and oxidative stress levels were determined by the DCF-DA reactive oxygenation assay.The mRNA expression levels of heme oxygenase-1(HO-1)mRNA and NAD(P)H:quinone oxidoreductase-1(NQO-1)were quantified using RT-qPCR.RESULTS:Treatment with varying concentrations of HMF resulted in a significant reduction in the proliferative capacity of SW480 and HCT116 cancer cells,as was indicated by CCK-8 experiments(P<0.05).Transwell assays demonstrated a pronounced attenuation in the invasive potential of SW480 and HCT116 following HMF treatment(P<0.05).Scratch assays highlighted a notable constraint on the migratory capabilities of SW480 and HCT116 after HMF treatment(P<0.05).DCF-DA staining revealed a substantial increase in reactive oxygen species(ROS)levels within SW480 and HCT116 cells after HMF treatment(P<0.05).Furthermore,RT-qPCR experiments elucidated that HMF markedly suppressed the mRNA expression of antioxidant genes HO-1 and NQO-1.CONCLUSION:HMF induces oxidative stress response in SW480 and HCT116 cells,consequently inhibiting their proliferation,invasion and migration.