The Study Of 5-FU And Its Derivatives On Radiosensitivity To Cell Lines SHEE And Strain Of Cancer Cell SHEEC

Objective: To investigate the effect inhibition rates of 5-FU and its derivatives on radiosensitivity to esophageal immortalized epithelial cell lines SHEE and cancerization cell lines SHEEC, and detect its molecular mechanism,provide clinical theoretical evidences for 5-FU and its derivatives on radiosensitivity to esophageal carcinoma.Methods:1. Collectioned oral capecitabine and S-1 serum before and after for cells drug sensitivity test preparation.Detected the concentration of effective metabolites in serum (5-Fu, 5'-DFUR) with HPLC.2. The cell lines SHEE, SHEEC were divided into 36 groups, each group of 96 duplicate wells, used 0Gy, 0.5Gy, 1.0Gy, 2.0Gy, 4.0Gy, 6.0Gy different doses of rays were irradiated with each 3 groups, detected survival rates after 144h; by 2Gy ray irradiated with each 3 groups, detected survival rates after 24h, 48h, 72h, 96h, 120h, 144h.3. The cell lines SHEE, SHEEC were divided into 15 groups, each group of 96 duplicate wells, each 3 groups were given capecitabine serum, 5'-DFUR, S-1 serum, 5-Fu enhanced sensitivity and the control group, and then gived 2Gy irradiatied, detected inhibition rates after 144h of cell lines SHEE, SHEEC.4. The cell lines SHEE, SHEEC were divided into 18 groups, each group of 6 duplicate wells, each of 3 groups with capecitabine serum, S-1 serum enhanced sensitivity and the control group, and then gived 2Gy irradiatied, detected expression rates of apoptotic protein Bax, Bcl-2 after 144h of cell lines SHEE, SHEEC with immunofluorescence.Results: 1. 5'-DFUR and 5-Fu were not detected in oral capecitabine and S-1 serum.The concentrations of 5-Fu and 5'-DFUR after oral capecitabine serum was 101ng/ml, 2763ng/ml; The concentration of 5-Fu after oral S-1 serum was 4936ng/ml.2. After exposured irradiation dose 0.5 Gy, 1.0 Gy, 2.0 Gy, 4.0 Gy, 6.0 Gy, after 144h the survival rates of cell lines SHEE were 93.62%, 87.68%, 80.12%, 55.23%, 41.07%, after 144h the cell lines SHEEC survival rates were 94.13%, 89.49%, 81.51%, 56.57%, 43.51%. 2 Gy dose exposured cell lines SHEE afrer 24h, 48h, 72h, 96h, 120h, 144h, the survival rates were 114.15%, 130.65%, 143.67%, 89.53%, 85.48%, 80.02%, did the same the cell lines SHEEC after 24h, 48h, 72h, 96h, 120h, 144h. The survival rates were 115.24%, 131.58%, 145.50%, 90.56%, 86.71%, 80.53%.3. The radiosensitization effects of Cpecitabine seurm , 5'-DFUR, S-1 seurm, 5-Fu to the cell lines SHEE and SHEEC.(1) The radiation group compared with concentration of 10% capecitabine serum after sensitization 144h to SHEE cells inhibition rates were 32.67%, 35.21%, the statistically was no significant differences (P>0.05); to SHEEC cells after sensitization 144h inhibition rates were 36.32%, 51.62%, the statistically was significant differences (P<0.05); to SHEE and SHEEC cells after sensitization 144h inhibition rates were 35.21%, 51.62 %, the statistically was significant differences(P<0.05).(2) The radiation group compared with concentration of 276.3ng/ml of 5'-DFUR (10% concentration of capecitabine serum of 5'-DFUR) after sensitization 144h to SHEE cells inhibition rates were 35.23%, 37.64%, the statistically was no significant differences (P>0.05); to SHEEC cells after sensitization 144h inhibition rates were38.65%, 54.21%, the statistically was significant differences(P<0.05); to SHEE and SHEEC cells after sensitization 144h inhibition rates were37.64%, 54.21%, the statistically was significant differences(P<0.05).(3) The radiation group compared with concentration of 10% S-1 serum after sensitization 144h to SHEE cells inhibition rates were 31.62%, 55.37%, the statistically was significant differences (P<0.05); to SHEEC cells after sensitization 144h inhibition rates were 32.61%, 52.31%, the statistically was significant differences (P<0.05);to SHEE and SHEEC cells after sensitization 144h inhibition rates were 55.37%, 52.31%, the statistically was no significant differences (P>0.05).(4) The radiation group compared with concentration of 493.6ng/ml of 5-Fu (10% concentration of S-1 serum of 5-Fu ) after sensitization 144h to SHEE cells inhibition rates were 31.32%, 55.13%, the statistically was significant differences (P<0.05); to SHEEC cells after sensitization 144h inhibition rates were 32.51%, 50.27%, the statistically was significant differences (P<0.05); to SHEE and SHEEC cells after sensitization 144h inhibition rates were 55.13%, 50.27%, the statistically was no significant differences (P>0.05).4. The expression effects of Capecitabine,S-1 serum to the protein Bcl-2,Bax(1) The radiation group compared with concentration of 10% capecitabine serum to cell lines SHEE after 144h, the expression rates of Bcl-2 protein were 25%, 23%, the statistically was no significant differences (P>0.05) the expression rates of Bax protein were 26%, 28%, the statistically was no significant differences (P>0.05); to cell lines SHEEC after 144h, the expression rates of Bcl-2 protein were 26%, 15%, the statistically was significant differences (P<0.05), the expression rates of Bax protein were27%, 52%, the statistically was significant differences (P<0.05). The radiation group compared with concentration of 10% capecitabine serum to cell lines SHEE and SHEEC after 144h, the expression rates of Bcl-2 protein were 23%, 15%, the statistically was significant differences (P<0.05), the expression rates of Bax protein were 28%, 52%, the statistically was significant differences (P<0.05).(2) The radiation group compared with concentration of 10% S-1 serum to cell lines SHEE after 144h, the expression rates of Bcl-2 protein were 24%, 12%, the statistically was significant differences (P<0.05), the expression rates of Bax protein were 25%, 51%, the statistically was significant differences (P<0.05), to cell lines SHEEC after 144h, the expression rates of Bcl-2 protein were 26%, 14%, the statistically was significant differences (P<0.05), the expression rates of Bax protein were 25%, 53%, the statistically was significant differences (P<0.05). The radiation group compared with Concentration of 10% S-1 serum to cell lines SHEE and SHEEC after 144h, the expression rates of Bcl-2 protein were 12%, 14%, the statistically was no significant differences (P>0.05), the expression rates of Bax protein were 51%, 53%, the statistically was no significant differences (P>0.05).Conclusion:1. The main effective components of capecitabine serum was 5'-DFUR; The main effective components of S-1 serum was 5-FU.2. The best dose ray was 2Gy, the best detection of survival rates was sensitization after 144h. for cell lines SHEE, SHEEC.3. The study of radiosensitization (1) Capecitabine could significantly enhanced the radiation inhibition to SHEEC cell lines, but was insignificant sensitized effect to SHEE cell lines.(2) The radiosensitization of 5'-DFUR on the cell lines SHEE and SHEEC was same to the capecitabine results.(3) S-1 was significantly sensitized effect to both cell lines SHEE and SHEEC,it hadn't slective sensitization between normal cells and cancerization cells.(4) The radiosensitization of 5-FU on the cell lines SHEE and SHEEC was same to the S-1 results.Capecitabine, 5'-DFUR may be more suitabled for esophageal cancer radiosensitization.4. The study of apoptosis protein expression(1) Capecitabine may be the major injury of apoptosis in radiosensitivity to cell lines SHEEC, but less injury of apoptosis in radiosensitivity to cell lines SHEE.(2) S-1 may be injury of apoptosis in radiosensitivity both to cell lines SHEE and SHEEC.