DNA-PK的活性及蛋白表达与鼻咽癌细胞株CNE1CNE2放射敏感性的关系.pdfVIP

524 Acta Physiologica Sinica , August 25, 2007, 59 (4): 524-533 Research Paper DNA-dependent protein kinase activity and radiosensitivity of nasopharyngeal carcinoma cell lines CNE1/CNE2 1,2 1,2 1,2 1,2 1 1 1,2,* HE Yu-Xiang , ZHONG Ping-Ping , YAN Shan-Shan , LIU Li , SHI Hong-Liu , ZENG Mu-Sheng , XIA Yun-Fei 1State Key Laboratory of Oncology in Southern China; 2Department of Radiation Oncology, Cancer Center, Sun Yat-Sen University, Guangzhou 510060, China Abstract: The present study investigated the relationship between DNA-dependent protein kinase (DNA-PK) and radiosensitivity of nasopharyngeal carcinoma (NPC) cell lines. The dose-survival relationship for NPC cell lines, CNE1 and CNE2, was analyzed using clonogenic formation assay, the activity of DNA-PK of the two cell lines was measured using the Signa TECT DNA-PK assay kit, and the localization and expression of Kus (a heterodimer) and DNA-PKcs protein in CNE1 and CNE2 before irradiation and 15 min, 1 h, 6 h, 12 h, 24 h after 4 Gy irradiation were analyzed by immunofluorescence, laser scanning confocal microscope (LSCM) and Western blot. The results showed that the surviving fraction of CNE1 was higher than that of CNE2 at each dose. The DNA-PK activity of CNE1 was also significantly higher than that of CNE2 before and after irradiation (P0.05), while the expression of total Ku70/Ku80 in CNE1 and CNE2 had no significant difference. Increasing translocation of Ku70 and Ku80 from the cytoplasm to the nuclei in the two cell lines was observed with increase of irradiation time as detected by Western blot, and the immunofluorescence of the DNA-PK complex subunits showed greater nuclear translocation in CNE1 tha