鼻咽癌CNE1放射抗拒细胞亚系CNE1/70R与亲代细胞的差异

Differences between radioresistant cell subline CNE1/70R of nasopharyngeal carcinoma CNE1 and its parental cell line

目的探讨鼻咽癌CNE1细胞及放射抗拒细胞亚系CNE1/70R细胞在倍增时间、放射敏感性、药物敏感性、细胞凋亡、细胞周期方面的差异。方法用临床常规分割根治剂量70 Gy体外照射建立鼻咽癌放射抗拒细胞模型CNE1/70R。采用生长曲线检测CNE1及CNE1/70R两株细胞倍增时间,细胞克隆形成实验比较细胞放射敏感性,MTT法检测细胞对药物多西紫杉醇、顺铂的敏感性,流式细胞术检测细胞凋亡和周期的变化。结果 CNE1/70R和CNE1细胞的倍增时间分别为(1.612±0.069)、(1.481±0.046)d;与CNE1细胞比较,CNE1/70R细胞的放射抗拒性增强,对化疗药物多西紫杉醇和顺铂敏感性增高;细胞经照射后,CNE1细胞G_2/M期阻滞较CNE1/70R细胞更加明显;细胞经照射相同剂量后,CNE1细胞凋亡率高于CNE1/70R细胞。以上观察指标比较,两株细胞差异均有统计学差异(P均<0.05)。结论成功建立人鼻咽癌放射抗拒株CNE1/70R,且与亲代细胞在放射及药物治疗敏感性方面有稳定差异。

Objective To investigate the differences in the doubling time,radiosensitivity,drug sensitivity,apoptosis and cell cycle distribution between nasopharyngeal carcinoma CNE1 cells and radioresistant cell subline CNE1 /70 R.Methods Using a radical cure dose（ 70 Gy） of clinical routine segmentation in vitro irradiation to establish radioresistant nasopharyngeal carcinoma cell line model CNE1 /70 R. The doubling time of the two cell lines CNE1 and CNE1 /70 R was measured by cell growth curve. The radiobiological characteristics of the two cell lines were analyzed by the colony forming assay. MTT was used to test the drug sensitivity of the two cell lines to docetaxel and cisplatin. The apoptosis and cell cycle was analyzed by flow cytometry. Results The doubling time of CNE1 /70 R and CNE1 R cells was respectively（ 1. 612 ±0. 069） d and（ 1. 481 ± 0. 046） d. The result of colony formation assay showed that the radioresistance of CNE1 /70 R cells was stronger than that of CNE1. Compared with CNE1 cells,CNE1 /70 R cells were more sensitive to the drugs. After irradiation,the CNE1 cells blocked in the G_2/ M phase were more than those of CNE1 /70 R cells,CNE1 arrested more obvious than CNE1 /70 R. After the same dose of irradiation,the apoptosis rate of CNE1 cells was higher than that of CNE1 /70 R cells. Significant differences were found in the above indexes between these two cell lines（ all P 0. 05）. Conclusions We successfully establish radiaoresistant nasopharyngeal carcinoma cell subline CNE1 /70 R,and it has stable difference with its parental cell line in radiosensitivity and drug sensitivity.