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目的:建立人鼻咽癌放射抗拒细胞株CNE-2R,为研究鼻咽癌放射抗拒机制提供具有可比性的成对细胞。方法:用γ射线反复照射CNE-2细胞,筛选出放射抗拒细胞株CNE-2R;检测细胞生长的倍增时间;应用细胞克隆形成实验检测细胞的放射敏感性;MTT法检测照射后不同时间的存活分数;用流式细胞术检测细胞的周期分布特征。结果:CNE-2R的细胞群体倍增时间长于亲代CNE-2(4.5dvs3.6d),P=0.047;CNE-2及CNE-2R的放射敏感性明显不同,CNE-2R的放射抗拒性增加;照射后12hCNE-2R的存活率大于CNE-2(89.0%vs71.7%),P=0.001;细胞分布周期改变,CNE-2R的S期比例增高,G0/G1和G2/M期细胞减少,χ2=7.312,P=0.034;CNE-2R在照射后12～36h出现明显的G2/M期阻滞,亲本CNE-2仅在照射后12～24hG2/M期略有增加。结论:人鼻咽癌细胞株CNE-2经间歇性大剂量γ射线多次照射后得到的CNE-2R具有稳定放射抗拒性,并且显示出与亲代不同的细胞周期特征。
OBJECTIVE: To establish a human nasopharyngeal carcinoma cell line CNE-2R which can provide a comparable pair of cells for the study of radioresistance mechanism of nasopharyngeal carcinoma. Methods: The CNE-2 cells were irradiated with γ-ray repeatedly, and the cell line CNE-2R was screened out. The doubling time of cell growth was detected. The cell radiosensitivity was detected by cell clone formation assay. The survival of the cells was detected by MTT assay The cell cycle distribution was detected by flow cytometry. Results: The population doubling time of CNE-2R was longer than that of CNE-2 (4.5d vs. 3.6d), P = 0.047. The radiosensitivity of CNE-2 and CNE-2R was significantly different and the radioresistance of CNE-2R was increased. After 12h, the survival rate of CNE-2R was higher than that of CNE-2 (89.0% vs 71.7%), P = 0.001. The cell cycle distribution was changed, the proportion of S phase of CNE-2R increased and the number of cells in G0 / G1 and G2 / = 7.312, P = 0.034; CNE-2R showed significant G2 / M arrest at 12-36h after irradiation, while parent CNE-2 increased slightly only at 12-24h G2 / M after irradiation. CONCLUSION: CNE-2R obtained after repeated irradiation of human nasopharyngeal carcinoma cell line CNE-2 by intermittent high-dose γ-rays has stable radioresistance and shows different cell cycle characteristics than the parental.