Quan Wen1, Yu-pei Zhao, Ge Chen, Fu-quan Zhang, Tai-ping Zhang. 1. Department of General Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China.
Abstract
OBJECTIVE: To investigate the association of S100A2 expression with the secondary radioresistance in pancreatic cancer cells. METHODS: Human pancreatic cancer cells of the line SW1990, parental line, and the secondarily radioresistant human pancreatic cancer cells of the line SW1990/R were cultured and inoculated into the wells of a 6-well plate. 5-Aza-2'-deoxycytidine (5-aza-DC) of different concentrations was added into the culture fluid. Forty-eight hours later X-ray irradiation was conducted. Cloning efficiency assay was used to assess the cell survival. The cell cycle was analyzed by flow cytometry. The mRNA expression of S100A2 was examined by real-time RT-PCR and the protein expression of S100A2 was examined by immunofluorescence quantitative analysis. RESULTS: The S100A2 mRNA expression level of the SW1990 line was significantly higher than that of the SW1990/R line. Treated by 5-aza-dC, the S100A2 mRNA expression level of the SW1990/R line increased by 2 approximately 3 times. The S100A2 protein expression level of the SW1990 line was significantly higher than that of the SW1990/R line (t = 3.648, P = 0.022). Treated by 5-aza-dC, the S100A2 protein expression level of the SW1990/R line increased, by 5 times at most (t = -11.404, P = 0.001). The G(2)/M of the SW1990 line was 16.2%, significantly higher than that of the SW1990/R line (16.2%), and the G(2)/M levels of the SW1990/R line treated by 5-aza-dC of different concentrations were all increased dose-dependently. The colony-forming rates of the SW1990/R line of the 5-aza-dC + irradiation groups were all significantly lower than that of the SW1990/R line of the pure irradiation group, even the dose of 5-aza-dC was as low as 0.025 microm. The SF2 (cell survival rate with the irradiation dose of 2 Gy) of the 5-aza-dC + irradiation group was 0.122, significantly lower than that of the pure irradiation group (0.460). CONCLUSION: The expression level of S100A2 is low in the radioresistant pancreatic cancer cells, which may be one of the mechanisms of radioresistance of these cells.
OBJECTIVE: To investigate the association of S100A2 expression with the secondary radioresistance in pancreatic cancer cells. METHODS:Humanpancreatic cancer cells of the line SW1990, parental line, and the secondarily radioresistant humanpancreatic cancer cells of the line SW1990/R were cultured and inoculated into the wells of a 6-well plate. 5-Aza-2'-deoxycytidine (5-aza-DC) of different concentrations was added into the culture fluid. Forty-eight hours later X-ray irradiation was conducted. Cloning efficiency assay was used to assess the cell survival. The cell cycle was analyzed by flow cytometry. The mRNA expression of S100A2 was examined by real-time RT-PCR and the protein expression of S100A2 was examined by immunofluorescence quantitative analysis. RESULTS: The S100A2 mRNA expression level of the SW1990 line was significantly higher than that of the SW1990/R line. Treated by 5-aza-dC, the S100A2 mRNA expression level of the SW1990/R line increased by 2 approximately 3 times. The S100A2 protein expression level of the SW1990 line was significantly higher than that of the SW1990/R line (t = 3.648, P = 0.022). Treated by 5-aza-dC, the S100A2 protein expression level of the SW1990/R line increased, by 5 times at most (t = -11.404, P = 0.001). The G(2)/M of the SW1990 line was 16.2%, significantly higher than that of the SW1990/R line (16.2%), and the G(2)/M levels of the SW1990/R line treated by 5-aza-dC of different concentrations were all increased dose-dependently. The colony-forming rates of the SW1990/R line of the 5-aza-dC + irradiation groups were all significantly lower than that of the SW1990/R line of the pure irradiation group, even the dose of 5-aza-dC was as low as 0.025 microm. The SF2 (cell survival rate with the irradiation dose of 2 Gy) of the 5-aza-dC + irradiation group was 0.122, significantly lower than that of the pure irradiation group (0.460). CONCLUSION: The expression level of S100A2 is low in the radioresistant pancreatic cancer cells, which may be one of the mechanisms of radioresistance of these cells.