The Effect Of Quercetin On Human Esophageal Cancer Cell Lines Eca109 Proliferation And Apoptosis And Its Possible M Echanism

Objective: Observe the effect of Quercetin on esophageal cancer cell lines Eca109’s proliferation and apoptosis, discuss the apoptosis mechanism of Quercetin effect on esophageal cancer cell lines Eca109.Methods:To foster esophageal cancer cell lines Eca109 in vitro, chose the logarithmic phase of esophageal Eca109 cells, digested by trypsin, made into single cell suspension, set five different concentration group of quercetin(0μmol/L,40μmol/L, 80μmol/L, 160μmol/L, 320 μmol/L), each group set five parallel hole. Using standard cell count plate count, adjust the cell concentration on 96-well plate as 1*105 / ml, each hole to add 100 ml cell suspension. And 24 h, 48 h, 72 h later, observe its influence on Eca109 cell proliferation respectly. Detected by MTT method, use microplate reader to measure the OD value of each hole, according to the OD value to calculate the of Eca109 cell inhibition rate, repeat the above experiments three times respectively, average the above inhibition rate, to calculate the different time, different concentra- tion proliferation inhibition rate of quercetin on human esophageal Eca109 cells. Continue to foster esophageal cancer Eca109 cells, as well as chose logarithmic growth phase cells, use trypsase digestion it into single cell suspension, PBS washing after centrifugal, centrifugal again and collecting cells. Added Binding Buffer, Annexin V- FITC, PI dyeing liquid respectively, detect cell apoptosis rate. Using flow cytometry to test apoptosis of different concentration of quercetin(0 μmol/L, 40 μmol/L, 80 μmol/L, 160μ mol/L, 320 μmol/L) effect on Eca109 24 h later. After get apoptosis rate, chose optimal concentration, use Western blot method to detect the content of apoptosis protein Bcl-2 and Bax and its expression. The tial result were resolveded by calculator software SPSS 19.0, regard P<0.05 as there were discrepancy difference.Results: Five different concentration of quercetin(0μmol/L,40μmol/L, 80μmol/L, 160μmol/L, 320 μmol/L) effect on human esophageal cancer Eca109 cell,24h, 48 h, 72 h later respectively measure the proliferation inhibition rate( 24h:0%, 26.3%, 36.0%, 52.8%, 64.4%;48h:0%, 27.3%, 37.8%, 54.7%, 63.8%;72h:0%, 28.0%, 39.7%, 58.5%, 66.9%). With the increase of concentration of quercetin, the extension of time, the inhibition effect increased(P < 0.05). The proliferation inhibition rate of different concentration of quercetin effect on human esophagus cancer Eca109 cell is significantly higher than control group(P < 0.05). Five different concentration of quercetin(0μmol/L,40μmol/L,80μmol/L, 160μmol/L, 320 μmol/L) effect on human esophageal cancer Eca109 cell, 24 h later, using flow cytometry to test apoptosis. 0μmol/L to320 μmol/L, the apoptosis inhibitory rate were: 2.09%, 6.87%, 14.7%, 19.4%, 22.3%. As the concentration of quercetin the increase, the apoptosis increased(P < 0.05); significantly higher than the control group respectly(P < 0.05). In the concentration of 320 μmol/L and 160 μmol/L group, using Western blot method to detect its apoptosis protein Bcl-2 and Bax, compare to the latter, the former express Bcl-2 decrease, express Bax increase, the difference was statistically significant(P < 0.05).Conclusion: Quercetin could significantly inhibit esophageal Eca109’s proliferation and induce cell apoptosis in a time and with dose dependent manner. Its mechanism may be related to the decreased express of protein Bcl-2 and the increased express of protein Bax.