人宫颈癌获得性放射抗拒细胞株(Hela-R)DNA修复能力的观察  

Observation of the DNA repair capability of acquired radioresistant cell line from human cervical carcinoma cells

作　　者：刘晨[1] 庞学利[1] 

机构地区：[1]第三军医大学西南医院肿瘤科,重庆400038

出　　处：《重庆医学》2013年第12期1370-1373,1376,共5页Chongqing medicine

摘　　要：目的观察人宫颈癌获得性放射抗拒细胞株(Heal-R)与其亲本细胞(Hela)DNA修复能力的差异。方法采用克隆形成实验测定细胞的放射敏感性;CCK-8细胞增殖实验检测增殖情况,流式细胞术检测凋亡及周期变化,彗星实验及5-乙基-2′-脱氧尿苷(EdU)掺入检测DNA的修复能力。结果 Hela-R细胞株的平均致死剂量显著高于Hela细胞株。X射线照射后,Hela-R细胞株早、晚期凋亡率均低于Hela细胞。24、48、72hHela和Hela-R细胞株的光密度值(OD)值分别为1.13±0.12、1.46±0.13(P<0.05);1.34±0.07、1.20±0.07(P<0.05);1.58±0.07、1.48±0.07(P<0.05)。Hela-R细胞株放疗后12～48hG2期细胞显著增多。Hela-R细胞株放疗后24h与48h,彗星尾长均短于Hela细胞株。X射线照射1h后,Hela-R细胞株EdU荧光强度为121.32±39.67(P<0.01),高于Hela细胞株。结论 Hela-R较Hela细胞株具有显著的放射抗拒性,DNA损伤修复能力的增强是其产生的重要机制之一。Objective To observe the difference of DNA repair capability between the radioresistant cell （Hela-R） line and its parents cell（Hela） line. Methods The radiosensitivity was measured with colony formation. CCK-8 cell proliferation assay was used to detect proliferation after radiation. Flow-cytometry was used to detect the change of apoptosis and ceil cycle after radiation. Comet assay and incorporation of ethynyl deoxyuridine were used to detect DNA repair capability. Results The mean lethal dose of Hela-R was significantly superior to its parental cell （Hela）. After irradiation, Hela-R cell line＇s early apoptotic rate and late apop-totic rate were significantly less than the Hela cells. CCK 8 proliferation experiment showed that Hela and Hela-R cell＇s OD value, respectively,in 24,48 and 72 hours,were 1.13 ± 0.12 and 1.46 ± 0.13 （P〈0.05） ;1.34 ± 0.07 and 1.20 ± 0.07 （P〈（0.05） ; 1.58 ± 0.07 and 1.48 ± 0. 07（P〈0. 05）. Hela-R＇s G2 phase rate was significantly higher than Hela in 24 to 48 hours after radia-tion. After irradiation, Hela-R＇s comet tail length were significantly shorter than the same period of Hela. In 1 hour after irradia- tion,incorporation of ethynyl deoxyuridine （EdU） showed that Hela-R＇ s fluorescence intensity were 121.32 4±39.67 （ P〈 0.01）. They were significantly higher than the same period of Hela. Conclusion The radioresistant effect is significant strong in Hela-R than Hela. The increase of DNA repair capability is an important mechanism of radioresistance after radiotherapy in recurrent cervi-cal cancer.

关 键 词：宫颈肿瘤 放射抗拒细胞株 DNA修复 

分 类 号：R737.33[医药卫生—肿瘤]