目标:山奈酚,一种天然类黄酮,大量发生在水果和蔬菜中。它具有多种生物活性,抗氧化剂,抗炎,和其他有益的特性。本研究的目的是研究山奈酚对体外增殖的影响。凋亡,KB细胞的自噬,人类宫颈癌细胞系,以及相应的作用机制。
方法:通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定法研究山奈酚对KB宫颈癌细胞的抑制作用,迁移测定,4\',6-二氨基-2-苯基吲哚染色,流式细胞术,吖啶橙染色和蛋白质印迹。
结果:山奈酚以剂量依赖性方式降低KB细胞活力和迁移。此外,山奈酚诱导的细胞凋亡得到证实,和山奈酚处理影响凋亡蛋白的水平。在特征性自噬液泡和酸性囊泡细胞器的可视化中检测到自噬,并使用西方印迹进行了验证,显示自噬相关蛋白水平升高。山奈酚介导的细胞凋亡和自噬明显归因于磷酸肌醇3-激酶(PI3K)/丝氨酸/苏氨酸激酶1(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)途径的磷酸化降低。使用PI3K途径抑制剂LY294002的药理学抑制试验验证了这一发现,PI3K途径抑制剂LY294002促进了KB细胞凋亡和自噬。
结论:我们的结果表明山奈酚通过抑制人宫颈癌细胞PI3K/AKT/mTOR通路诱导细胞凋亡和自噬,凭经验证明山奈酚的抗癌作用,并因此将其作为潜在的抗癌治疗剂。
OBJECTIVE: Kaempferol, a natural flavonoid, occurs abundantly in fruits and vegetables. It has various bioactivities, with antioxidant, anti-inflammatory, and other beneficial properties. The aim of this study was to investigate the in vitro effects of
kaempferol on the proliferation, apoptosis, and autophagy of KB cells, a human cervical cancer cell line, and the corresponding action mechanisms.
METHODS: The inhibitory efficacy of
kaempferol on KB cervical cancer cells was investigated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, migration assay, 4\',6-diamidino-2-phenylindole staining, flow cytometry, acridine orange staining and western blotting.
RESULTS: Kaempferol reduced KB cell viability and migration in a dose-dependent manner. Additionally,
kaempferol-induced apoptosis was confirmed, and
kaempferol treatment influenced levels of apoptotic proteins. Autophagy was detected upon visualization of characteristic autophagic vacuoles and acidic vesicular organelles, and verified using western blotting, which revealed elevated levels of autophagy-related proteins. Kaempferol-mediated apoptosis and autophagy were evidently attributable to reduced phosphorylation in the phosphoinositide 3-kinase (PI3K)/serine/threonine kinase 1 (AKT)/mammalian target of rapamycin (mTOR) pathway. This finding was validated using a pharmacological inhibition assay with the PI3K pathway inhibitor LY294002, which promoted KB cell apoptosis and autophagy.
CONCLUSIONS: Our results suggest that kaempferol induces apoptosis and autophagy by inhibiting the PI3K/AKT/mTOR pathway in human cervical cancer cells, empirically showing the anticancer effects of kaempferol, and thereby presenting it as a potential anticancer therapeutic agent.