Experimental Study On The Effects Of Rats Serum Of Long-term Vinegar Diet Inducing Apoptosis And Survivin Expression Of Lung Adenocarcinoma Cell Line A549

OBJECTIVESIn order to explore the anti-tumor effects of vinegar,this experiment simulated living conditions of the long-term vinegar diet.Rats serum that had been under the conditions of long-term vinegar diet,was used to culture A549 cell,directly at the cellular level studying on proliferation activity,apoptosis rate and Survivin expression of A549 cell.To confirm the inhibited effects of vinegar on A549 proliferation and explore primitively the possible mechanisms.METHODS1. Experimental Methods50 SD rats that were male and female in the same number were randomly divided into the group of long-term vinegar diet and the control group. The control group was fed by normal foods and water, but the long-term vinegar diet group was fed by the special foods and water, which were marinated by high concentration vinegar (vinegar: normal diet = 3:7, vigegar :normal water=1:20). Temperature was about 25±2℃in animal room, humidity was from 40% to 60%.Eight months later, separating serum from rats blood, serum was used to replace fetal bovine serum medium to culture human lung adenocarcinoma cell A549. The experiment were divided into four groups: experimental group (V group): Including 20% inactivated rats serum from long-term vinegar diet group ;control group (N group): Containing 20% inactivated serum of rats was fed by normal foods and water. D and NN group that separatively including 3 mg/L ShunBo and 10% fetal calf serum had been added through all experiment for a comparsion.2. Detection of experimental indicators MTT assay be used to determined the proliferation inhibition by the rats serum that long-term vinegar diet to A549.FCM was employed test the apoptosis rate of A549 cells induced by 20% rats serum that long-term vinegar diet. AnnexinⅤ-EGFP(+)P(I-)indicated the early apoptosis,but the Apo Peak illustration late apoptosis. Using Western-blot method to measure the expression level of Survivin protein of A549 in different groups.3. Statistic analysisAll experimental data was expressed by ( x±s). The results were analyzed by SPSS15.0. The minimum criterion for statistical significance was set at p<0.05.RESULTS1. The effect of long-term vinegar diet condition on human lung adenocarcinoma A549 cell proliferationCompared with the OD value that were measured at different time, revealed that among different groups there were significant differences ( p <0.05 ).Between V and N group there were significant differences ( p <0.05 ).The cell proliferation of V group is bad than N group. According to OD value in different time points, we can get growth curve. The inhibition in a time-dependent manner(r=0.980, p <0.05).2. The effect of long-term vinegar diet condition on human lung adenocarcinoma A549 cell apoptosis.To analyze the apoptosis rate that were measured by FCM revealed that different groups have significant differences ( p <0.05 ).The ratio between V and N group have significant differences ( p <0.05 ).The apoptosis rate of V group would be significantly higher than N group.3. The effect of long-term vinegar diet condition on A549 cell Surviving protein expression.Statistical analysis revealed that the difference among different group have significant ( p <0.05 ). Western-blot proved that the level of Survivin protein expression was markerly decrease in A549 cell treated with 20% rats serum from V group for 48 hour than N group. Conclusions1.Experimental result showed that rats'serum on long-term vinegar diet can inhibit the proliferation of lung cancer cells and appeared time-dependent inhibition.2.Rat's serum on long-term vinegar diet condition can inducing apoptosis of lung cancer cell,which is one of an important factor of inhibiting cell proliferation.3.Rat's serum on long-term vinegar diet condition can decrease Survivin protein expression.It is one of a mechnisms of inducing apoptosis probably.