纳秒电脉冲致人卵巢癌细胞DNA损伤的时间效应  

Temporal Pattern of DNA Breaks in Human Ovarian Cancer Cells after Exposure to Nanosecond Electric Pulses

作　　者：刘绍丽[1] 付晓[1] 任学毅 于廷和[1] 胡丽娜[1] 

机构地区：[1]重庆医科大学附属第二医院妇产科,重庆400010 [2]重庆市食品药品检验所,重庆401121

出　　处：《生物医学工程学杂志》2015年第5期1075-1078,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(31470822);高等学校博士学科点专项科研基金资助项目(20135503130002;20125503110004)

摘　　要：本研究旨在探讨纳秒电脉冲(nsEP)对人卵巢癌细胞铂敏感株A2780及铂耐药株C30的DNA损伤效应。将A2780和C30分别作用于场强为6kV/cm,脉宽为24ns的脉冲电场处理60s。假处理组予假脉冲处理。nsEP处理后0h、4h、8h、12h和24h采用CCK-8法检测细胞生存率;采用碱性单细胞凝胶电泳实验(彗星实验)检测细胞DNA损伤,测定彗星尾长(TL)、尾矩(TM)和Olive尾矩(OTM)的75th百分位数。结果显示随着时间增加,A2780和C30细胞的生存率和彗星形成率均先降低后增加,8h时最低(P<0.05);C30细胞的生存率高于A2780细胞(P<0.05),彗星形成率低于A2780细胞(P<0.05);C30细胞的TL、TM和OTM均低于A2780细胞(P<0.05);细胞死亡率和彗星形成率呈正相关(r=0.997,P<0.05;r=0.998,P<0.05)。结果表明,nsEP对人卵巢癌铂敏感细胞株和铂耐药细胞株的DNA损伤效应存在差异;nsEP致细胞死亡与DNA损伤相关。This study aims to explore the temporal pattern of DNA breaks induced by nanosecond electric pulses （nsEP） in cisplatin-sensitive and cisplatin-resistant human ovarian cancer cells. Human ovarian cancer cells A2780 （cisplatin-sensitive subline） and C30 （cisplatin-resistant subline） were exposed to nsEP. Sham exposed groups were shame exposed to nsEP. Cell viability was determined using CCK-8 assay after 0 h, 4 h, 8 h, 12 h and 24 h, respec- tively, and the percentage of dead cells was calculated. The DNA break was detected with the alkaline single cell gel electrophoresis （comet assay）, and the 75th percentiles of TL （tail length）, TM （tail moment） and OTM （Olive tail moment） were measured. Cell viability displayed an early decrease and late increase, with the valley value seen at 8 h. Percentages of cell death and comet-formed in A2780 cells were higher than those in C30 cells （P〈0.05） at 8 h, respectively. TL, TM and OTM in C30 cells were less than those in A2780 cells （P〈0.05）. The percentage of comet-formed correlated with that of cell death in either A2780 （r=0. 997, P%0.05） or C30 （r=0. 998, P〈0.05） cells. DNA breaks induced by nsEP in cisplatin-sensitive cells differred from that in resistant cells, and DNA break resulted in fraction of cell death.

关 键 词：纳秒电脉冲 卵巢癌 顺铂耐药 DNA损伤 

分 类 号：R737.31[医药卫生—肿瘤]