Study On Mechanisms Apoptosis Of MCF-7Cells Induced By Juglone In Mitochondrial Apoptotic Pathway

Juglone a compound of naphthoquinone is sourced from Juglans Mandshurica Maxim. Juglone has identified anti-tumor activity.But the mechanism of action had not identified. Still in the research stage of exploration. The study found that juglone rapidly oxidation Single-electron to semiquinone free radicals. in vivo, Semiquinone free radicals that can quickly make Dioxygen recovery to superoxide anion (O2-), which then recolonization juglone. the same time O2-under the action of the enzyme or the natural disproportionation reaction produce H2O2, The latter through the Fenton reaction, Then transformed into highly reactive hydroxyl radical(OH-), O2-、H2O2and OH-are reactive oxygen species(ROS).According to the literature, A variety of tumor cells apoptosis have the participation of reactive oxygen species, therefore, Reactive oxygen species in antineoplastic research has some theoretical significance and practical significance. The topic of this as a suitable solution for thorough exploration and research,and then to clarify whether juglone indirectly through reactive oxygen species-induced molecular mechanisms of tumor cell apoptosis, in order to further the juglone-based anti-cancer drug developed to provide some experimental bdecreased.MTT assay,inverted microscope investigate juglone for study of MCF-7cell growth inhibition, the results show that, with concentrations of juglone increasing, cell growth inhibition rate gradually increased, Inverted microscope to observe MCF-7cells, appeared suspended, and the higher the concentration of juglone, the role of the longer, more obvious, NAC interference group compared with juglone alone, growth inhibition was significantly reduced. Using Giemsa staining, the results show that, with increasing concentrations of juglone, MCF-7cells were dispersed, condensed cytoplasm, nuclear fragmentation, cell apoptotic bodies gradually increased; fluorescence microscopy, the results show that, with juglone concentration increased, the number of apoptotic bodies increased; with PI single staining observed under the effect of different concentrations of juglone apoptosis in MCF-7cells, the results show that, with increasing concentrations of juglone apoptosis also increased. NAC interference group compared with juglone alone, induction of apoptosis was significantly decreased. The use of flow cytometry analysis of the role of different concentrations of juglone MCF-7cells after24h, the level of reactive oxygen species and mitochondrial membrane potential changes, the results showed that, step-up levels of reactive oxygen species, descent mitochondrial membrane potential, NAC interference group and individual juglone group, Descent the level of reactive oxygen species, mitochondrial membrane potential rise; Laser confocal microscope, the juglone affect on MCF-7cells after24h of Ca2+concentration, the results show that, with increasing concentrations of juglone cells also will increase the concentration of Ca2+, NAC interference group compare with juglone alone group, the intracellular Ca2+concentration significantly decreasedTo sum up, Juglone can inhibit the proliferation of MCF-7cells and step-up MCF-7levels of intracellular ROS, oxidative damage to mitochondria, which leads to mitochondrial bulge,ROS through oxidation mitochondrial membrane, mitochondrial DNA and some important proteins, resulting function of mitochondrial electron transport chain occurs disabili-ty, resulting in decreased mitochondrial membrane potential, mitochondrial membrane permeability changes, cytochrome C and apoptosis correlation factor release. Calcium ion concentration increased, decreased mitochondrial membrane potential will lead to increased mitochondrial Ca2+uptake, mitochondrial calcium overload leading to mitochondrial damage, cyt-c release, activation of caspases, ultimately leading to apoptosis.