The Mechanism By Which Aesculetin Induces Apaptosis Of SGC-7901 Cells Through Regulating The Smac、Survivin Protein

Aesculetin is one of the component of medicinal materials cortex fraxini,researches show that Aesculetin are shown for a wide variety of tumor cells in vitro growth inhibition. This experiment selects the gastric cancer cell line SGC-7901,antitumor activity of Aesculetin and to study the anti-tumor mechanism of inducing cell apoptosis,Previous studies have shown that Aesculetin for some apoptosis factors influential in the mitochondrial apoptosis pathway, Such as the Bcl-2 family,cytochrome C and so on, But this experiment by Aesculetin of reactive oxygen species, calcium ions, the influence of the membrane potential, And its influence to the Smac, Survivin protein activity, further study on the anti-tumor mechanism. For the research of Aesculetin, anti-tumor mechanism to provide theoretical added. The clinical development of Aesculetin also has a c-ertain significance.Objective:Explore the Aesculetin effect on SGC-7901 cells, Reveals the mitochondrial apoptosis mechanism, Lays the foundation for subsequent research and exploration.Method:MTT method is used to observe Aesculetin on SGC-7901 cell growth inhibition, Fluorescence microscope showed, Using flow cytometry instrument analysis of different concentrations Aesculetin SGC-7901 cells after 24 hours, Reactive oxygen species level, mitochondrial membrane potential change and calcium ion concentration changes, The westen blot method at different concentrations Aesculetin effect after 24h, SGC-7901 cells the expression of Caspase-3, Caspase-9, Smac, Survivin protein. The results showed that with the increase of Aesculetin concentration, significantly increased the expression of Caspase-3, significantly increased the expression of Caspase-9, signifycantly increased the expression of Smac, while the expression of Survivin significantly reduced.Results:MTT method is used to observe Aesculetin on SGC-7901 cell growth inhibition, The results showed, With the increase of Aesculetin concentration, Cell growth inhibition rate increased. Fluorescence microscope showed. With the increase of Aesculetin concentration, The number of apoptotic body also will increase. Using flow cytometry instrument analysis of different concentrations Aesculetin SGC-7901 cells after 24 hours, Reactive oxygen species level, mitochondrial membrane potential change and calcium ion concentration changes, The results showed, Reactive oxygen species level rised, Mitochondrial membrane potential level declined, Calcium ion concentration increased. The westen blot method at different concentrations Aesculetin effect after 24h, SGC-7901 cells the expression of Caspase-3, Caspase-9, Smac,Survivin protein. The results showed that with the increase of Aesculetin concentration, significantly increased the expression of Caspase-3, significantly increased the expression of Caspase-9, signifycantly increased the expression of Smac, while the expression of Survivin significantly reduced.Conclusion:To sum up, Aesculetin can inhibit the SGC-7901 cells proliferation. and through the elevated level of reactive oxygen species, increased calcium ion concentration prompted mitochondrial permeability transition pore (MPTP) open, Lead to high osmotic pressure of mitochondrial matrix, so that the H+gradient disappeared, in turn, reduces the mitochondrial membrane potential, respiratory chain uncoupling, energy, water and solutes entering the mitochondrial matrix, break the outer membrane, and then release the apoptosis factor (Smac) protein, The positioning signal of Smac protein peptid-e is removed, resulting in Smac protein is released into the cytoplasm, Smac with Survivin (Survivin protein were found in recent years new apoptosis inhibiting protein, studies have shown that Survivin protein in many tumor cells increased,)can be specific binding, To remove Survivin protein of caspase inhibition, indirectly promoted the caspase apoptosis pathway activation, lead to apoptosis.