Xiao Chuang-Ying, Zhou Fu-Xiang, Liu Shi-Quan, Xie Cong-Hua, Dai Jing, Zhou Yun-Feng
Department of Radiochemotherapy, Zhongnan Hospital, Wuhan University, Wuhan, Hubei 430071, P. R. China.
Ai Zheng. 2005 Jun;24(6):653-6.
BACKGROUND & OBJECTIVE: Many studies showed that telomere length and telomerase activity closely correlate with proliferation and malignant degree of tumor cells, and both of them might be involved in the repair of radiation-induced DNA damage. So it was speculated that telomere length and telomerase activity maybe correlate to radiosensitivity of carcinoma cells. This study was designed to investigate the correlations of telomere length and telomerase activity to radiosensitivity of human laryngeal squamous carcinoma cell line Hep-2.
Hep-2 cells were irradiated with 0, 2, 4, 8, or 12 Gy of gamma-ray for 3 times. Survival cells were subcultured for 20 generations. Radiosensitivity index, survival fraction at 2 Gy (SF(2)), was measured by clone formation assay. Telomere length (mean length of telomere restriction fragments, TRF) was examined by Southern blotû telomerase activity (TA) was detected by polymerase chain reaction-based telomeric repeat amplification protocol (PCR-TRAP) coupled with ELISA.
After different doses of irradiation, SF(2) of Hep-2 cells was 0.47-0.64; TRF was 3.76-9.43 kb; TA was 1.761-2.606. Each parameter had significant differences among the survival progenies (P<0.05). SF(2) was positively correlated with TRF (r=0.921, P<0.01), and negatively correlated with TA (r=-0.929, P<0.01); TRF was negatively correlated with TA (r=-0.944, P<0.01).
Radiosensitivity of Hep-2 cells negatively correlates with telomere length, and positively correlates with telomerase activity, which suggest that both telomere length and telomerase activity may be used to predict cellular radiosensitivity.
多项研究表明,端粒长度和端粒酶活性与肿瘤细胞的增殖及恶性程度密切相关,二者可能均参与了辐射诱导的DNA损伤修复。因此推测端粒长度和端粒酶活性可能与癌细胞的放射敏感性相关。本研究旨在探讨端粒长度和端粒酶活性与人喉鳞状癌细胞系Hep-2放射敏感性的相关性。
用0、2、4、8或12 Gy的γ射线对Hep-2细胞照射3次。存活细胞传代培养20代。采用克隆形成试验测定放射敏感性指数,即2 Gy时的存活分数(SF(2))。通过Southern印迹法检测端粒长度(端粒限制片段平均长度,TRF);采用基于聚合酶链反应的端粒重复序列扩增法(PCR-TRAP)结合ELISA检测端粒酶活性(TA)。
不同剂量照射后,Hep-2细胞的SF(2)为0.47 - 0.64;TRF为3.76 - 9.43 kb;TA为1.761 - 2.606。各参数在存活子代间差异有统计学意义(P<0.05)。SF(2)与TRF呈正相关(r = 0.921,P<0.01),与TA呈负相关(r = -0.929,P<0.01);TRF与TA呈负相关(r = -0.944,P<0.01)。
Hep-2细胞的放射敏感性与端粒长度呈负相关,与端粒酶活性呈正相关,提示端粒长度和端粒酶活性均可用于预测细胞的放射敏感性。