Study On Apoptosis Induced By Aesculetin Of SGC-7901 Tumor Cells

Aesculetin is a common natural antioxidants in Chinese cortex fraxini, Aesculetin play a important pharmacological action in chemical components of cortex fraxini, recent studies showed that Aesculetin had anti-tumor activity, and its anti-tumor mechanisms are not specific know yet, vitro methods is used to study anti-tumor role of aesculetin in this article. Assumed that aesculetin through the induction of apoptosis by death receptor pathway in its anti-tumor activity. gastric cancer cell SGC-7901 may be take as a modle of cancer desease. Apply the knowledge of molecular biology and immunology experimental, through the apoptosis experiments of tumor cells, clarified its anti-tumor action and mechanism for further study. Which provide a theoretical basis, and also a certain significance elements of the clinical development.MTT assay and SRB assay were used to investigate the action of growth inhibiting of aesculetin on on human gastric carcinoma SGC-7901 cells, the results showed that aesculetin can inhibit the growth of SGC-7901 cells, and the results are dosage-dependent.after 48h treated with aesculetin, IC50 of aesculetin examined by MTT assay was 0.54mmol-L-1;and GI50 of aesculetin examined by SRB assay was 0.286mmol-L-1. Use Transmission electron microscopy, the ultrastructure of SGC-7901 cells were observed the result showed that after treated with aesculetin for 48h at the dose of 0.28mmol-L-1,0.56mmol-L-1,1.12m mol-L-1, such as chromatin condensation, margination against nuclear envelope and formation of apoptotic bodies appears. FCM was used to observe the apoptosis rate of SGC-7901 cells. The results show that, after treated with aesculetin for 48h at the dose of 0.28mmol·L-1, 0.56mmol·L-1,1.12m mol·L-1, Every dose apoptosis rates of SGC-7901 cells were respectively 14.20%,31.28%,54.13%, apoptosis rate increased in a dose-dependent manner.Based on the above experiment, we studied on the pathway of aesculetin induced apoptosis in SGC-7901 cells. The study of mechanism in human gastric cancer cells SGC-7901 induced apoptosis by aesculetin, using immunohistochemical detection to study the effect of aesculetin on death receptor pathway protein Fas, FasL, FADD.the using the microplate reader to detect the activity expression of Caspase-8, Caspase-3. The results showed that the three doses of aesculetin (0.28mmol-L-',0.56mmol-L-1 and 1.12mmol-L-1) in role of SGC-7901 cells after 24h, Fas protein expression rate was 37.28%,59.70%,85.23%, with control group the difference was significant (P<0.01); FasL protein expression rates were 40.11%,60.38%, 87.23%, with control group the difference was significant (P<0.01); FADD protein expression rates were 36.98%,52.34%,86.64%, compared with control group significant difference (P<0.01). These results indicate that with the aesculetin dose raise, the Fas, FasL, FADD protein expression increased, and has a dose dependent manner. Caspase-8 activity were detected by microplate reader, the OD values were 0.118,0.288,0.345 (P<0.01);the OD values of Caspase-3 activity were 0.141,0.234,0.338 (P<0.01); that with the role of aesculetin the intracellular Caspase-8 and Caspase-3 activity was increased. And between the three doses a dose dependent manner.In summary, aesculetin could efficiently induce proliferation inhibition and apoptosis in SGC-7901 cells. Aesculetin apoptosis induced by tumor cells may promote the death receptor pathway through Fas, FasL, FADD protein expression, and the formation of aggregates for Caspase-8, Caspase-3 expression, which induced apoptosis of SGC-7901 cells. The mechanism of the apoptosis in SGC-7901 cells maybe in the death receptor pathway by promoted Fas, FasL, FADD protein expression,which formated polymer and activated the expression of Caspase-8, Caspase-3 that induced apoptosis in SGC-7901 cells.