Effects Of Reservatrol On Cell Cycle, Apoptosis And Expression Of Apoptosis-related Gene Pdcd5in786-0Cells

Objective To investigate the effects of reservatrol on the expression of PDCD5mRNA、protein of786-0human kidney cancer cells and786-0cell proliferation, apoptosis, and cell cycle.Methods Human renal carcinoma786-0cells which be recoveried and cultured in vitro to logarithmic phase passage be divided into different experiment groups. Appling the reservatrol (12.5μmol/L、25μmol/L、50μmol/L、100μmol/L) affectted on786-0human renal cell24h,48h, and72h. And set up blank control groups(medium control group). The changes in cell morphology was observed under the microscope The effect of Resveratrol on the expression of apoptosis-related protein PDCD5was analyzed by using Immunocyto chemical detection methods; The effect of Reservatrol on the mRNA level expression of RCC786-0PDCD5was analyzed by using In situ hybridization method;the cell cycle phase distribution and the cell apoptosis of human kidney cancer786-0cells were analyzed by using flow cytometry and TUNEL assay.Results1. Under an inverted microscope observation of human renal cell carcinoma786-0cells,we found:Normal the renal cell carcinoma786-0cell growth was good, spindle-shaped, round or polygonal cell outline was clear. The renal cell carcinoma786-0cells number significantly reduced,cell volume was reduced, cell nucleus narrow, rounded after treatment by Res for24h.2. Immunocytochemistry results:The expression of PDCD5protein started to rise after four different concentrations of resveratrol(12.5μmol/L,25μmol/L,50μmol/L,100μmol/L) dealing with human carcinoma786-0cells at24h,48h,72h. Expressions of PDCD5protein levels of25μmol/L resveratrol for24h was2.90±0.13,3.08±0.21,3.23± 0.21compared to the control group (1.10±0.19,1.13±0.21,1.42±0.25),the difference was significant (P<0.05). Continued to increase in medicine concentration the expression of PDCD5protein differences was not statistically significant (P>0.05), and when the reaction time reached24hours, continued to increase the role of time,the differences of the Expressions of PDCD5protein increased was no significant (P>0.05).3. In situ hybridization:The expression of PDCD5mRNA started to rise after four different concentrations of resveratrol(12.5μmol/L,25μmol/L,50μmol/L,100μmol/L) dealing with human carcinoma786-0cells at Id,2d,3d. Expressions of PDCD5mRNA levels of25μmol/L resveratrol for24h was3.35±0.23,3.40±0.31,3.60±0.26compared with the control group (1.47±0.27,1.78±0.38,1.88±0.31)(P<0.05). And when the reaction time reached24hours, continued to increase the reaction time, the differences of PDCD5mRNA expression had no significant (P>0.05).4. Flow cytometry to detect the cell cycle changes:The human786-0cells treated by12.5μmol/L,25μmol/L,50μmol/L,100μmol/L of resveratrol (RES) after24h, the ratio of cells in G1phase of25μmol/L resveratrol group was65.28±3.10, the percentage of cells in G2phase was9.67±0.43,compared with blank control group, the proportion of cells:75.23±2.58,11.35±2.37declined, and the difference was statistically significance (P<0.05). The ratio of S phase cells of25μmol/L resveratrol treated was24.79±3.72compared with blank control group, the proportion of cells13.17±1.36increased, the difference was statistically significant (P<0.05). Continued to rise the medicine concentration,the differences were no significant between the groups (P>0.05).5. TUNEL assay:The apoptosis ratio of786-0cells in25μmol/L resveratrol group was:6.01±0.59, the proportion of apoptosis compared with the control group:0.73±0.25declined, and the difference was statistically significant (P<0.05). When the role of resveratrol reached24hours, continued to increase the reaction time, the apoptosis ratio of human786-0cells in the resveratrol treatment group was:5.90±0.50,5,97±0.25compared with blank control group, the proportion of apoptosis:0.90±0.20,1.10±0.20 increased significantly, the difference was statistically significant (P<0.05), compared with6.01±0.59, the difference was not significant (P>0.05).Conclusions1. Resveratrol could increase the PDCD5protein and mRNA expression in human renal cell carcinoma.2. The human renal cell carcinoma786-0cells could be Induced apoptosis by resveratrol3. After the human renal carcinoma786-0cells which treated by resveratrol, the cell cycle had changed:the S phase cell ratio increased and the G1、G2phase decreased.