DNA损伤应答靶向抑制剂对卵巢癌细胞的化疗增敏作用  

Employing DNA Damage Response Inhibitors to Enhance Chemosensitivity of Ovarian Carcinoma Cells

作　　者：蔡思源[1,2] 唐子执[2] 曾鸣[2] 王小军[2] 楼江燕[1] 刘聪[2] 陈杰[1] 

机构地区：[1]四川大学华西第二医院妇产科,成都610041 [2]四川大学出生缺陷与相关妇儿疾病教育部重点实验室,成都610041

出　　处：《四川大学学报（医学版）》2016年第3期316-320,336,共6页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅应用基础计划项目(No.2012JY0011);四川省杰出青年学术技术带头人资助计划(No.2012JQ0005)资助

摘　　要：目的观察DNA损伤应答抑制剂及其联合常规化疗药物(顺铂等)在卵巢癌耐药细胞株OVCAR-8中的作用,研究其化疗致敏效应。方法利用针对DNA损伤应答关键信号蛋白质的抑制剂,与顺铂等连用处理卵巢癌细胞,分析这些药物处理方式对卵巢癌细胞的杀伤能力。MTT法检测不同药物作用后OVCAR-8的增殖抑制情况;免疫荧光法检测OVCAR-8中磷酸化组蛋白2A变体(γH2AX)和p53结合蛋白1(53BP1)的表达,观察二者在DNA损伤位点的募集和形成灶点的能力。结果毛细血管扩张共济失调突变蛋白(ATM)/ATM和Rad 3相关蛋白(ATR)抑制剂与顺铂联用能抑制损伤修复机制的活化,明显减弱OVCAR-8细胞的增殖活力(P<0.01),促进其凋亡;在羟基脲和Wortmannin联合处理时,OVCAR-8细胞的ATR转导信号(如γH2AX)减弱,细胞生存率明显降低(P<0.05);多聚ADP-核糖聚合酶(PARP)抑制剂与顺铂联合处理OVCAR-8细胞未发现明显的增敏作用(P>0.05)。结论适当的DNA损伤应答抑制剂有潜力提高常规化疗药物的抗肿瘤效果,以达到快速清除肿瘤细胞和防止产生耐药性的效果。Objective To assess the sensitisation effects of DDR inhibitors combined with conventional chemotherapeutics agents（cisplatin et al）in a drug-resistant ovarian cancer cell line（OVCAR-8）.MethodsInhibitors of DDR regulators with cisplatin were applied to challenge OVCAR-8,and evaluated the DNA damage response（DDR）and cytotoxic effects of different combination of chemicals.Inhibition of proliferation to OVCAR-8of different drugs was evaluated by MTT assay.The activation of phosphorylation of histone family 2A variant（γH2AX）and p53 binding protein 1（53BP1）in OVCAR-8were evaluated by immunofluorescence to observe their ability of recruitment and forming foci at DNA damage site.Results We observed that combined treatment of ataxia-telangiectasia mutated（ATM）/ATM and Rad 3-related（ATR）inhibitor and cisplatin can suppress the activation of damage repair mechanisms and weakened the proliferative activity of OVCAR-8cells（P〈0.01）;ATR pathway was suppressed and the signal ofγH2AX weakened and cell survival rate significantly reduced when combination therapy of HU and Wortmannin（P〈0.05）;poly ADP-ribose polymerase（PARP）inhibitor could not enhance chemosensitivity in OVCAR-8 cells when combined with cisplatin（P〈0.05）. Conclusion We substantiated that appropriate inhibitors of DNA damage response may have a potential to improve the anti-tumor effect of conventional chemotherapy drugs and prevent drug resistances.

关 键 词：卵巢肿瘤 顺铂耐药 DNA损伤应答 抑制剂 

分 类 号：R737.31[医药卫生—肿瘤]