中国癌症杂志 ›› 2021, Vol. 31 ›› Issue (2): 100-107.doi: 10.19401/j.cnki.1007-3639.2021.02.003

• 论著 • 上一篇    下一篇

Ku86通过调控TOP1和COPS5影响上皮性卵巢癌细胞对于顺铂的化疗敏感性

马 倩 1,2* ,刘懿萱 1 ,开今言 1 ,郭 林 1 ,卢仁泉 1   

  1. 1. 复旦大学附属肿瘤医院检验科,复旦大学上海医学院肿瘤学系,上海 200032 ;
    2. 西安交通大学第一附属医院检验科,陕西 西安 710000
  • 出版日期:2021-02-28 发布日期:2021-03-02
  • 通信作者: 卢仁泉 E-mail: lurenquan@126.com
  • 基金资助:
    国家自然科学基金(81572552,81772774,81772808);上海市科学技术委员会引导项目(17411963500,17411951000)。

Down-regulation of Ku86 inhibits cisplatin-induced chemotherapy sensitivity by regulating TOP1 and COPS5 in epithelial ovarian cancer

MA Qian 1, 2* , LIU Yixuan 1 , KAI Jinyan 1 , GUO Lin 1 , LU Renquan 1    

  1. 1. Department of Clinical Laboratory, Fudan University Shanghai Cancer Center; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; 2. Department of Clinical Laboratory, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710000, Shaanxi Province, China 
  • Published:2021-02-28 Online:2021-03-02
  • Contact: LU Renquan E-mail: lurenquan@126.com

摘要: 背景与目的:卵巢癌是妇科恶性肿瘤中发病率排名第3、致死率排名第1的疾病。卵巢癌的预后很差,这是由于大部分患者确诊时已处于晚期并且对铂类药物化疗易耐药,异常的DNA修复是导致铂类耐药的重要原因,针对性地干扰DNA损伤修复相关分子可能是提高铂类药物化疗敏感性的新手段。Ku86是参与非同源末端连接(non-homologous end joining,NHEJ)过程的一个关键分子,能够有效地修复DNA双链断裂(DNA double-strand break,DSB)。方法: 使用免疫组织化学染色和细胞免疫荧光法检测Ku86在卵巢癌组织和细胞中的定位。使用RNAi技术下调Ku86,用顺铂处理后使用流式细胞术检测凋亡,使用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测IC 50 。使用蛋白质印迹法(Western blot)分别检测并分析Ku86与拓扑异构酶Ⅰ(topoisomeraseⅠ,TOP1)及COP9信号复合体的第5种成分(the fifth component of the COP9 signalosome,COPS5)的关系。结果:下调Ku86可以减少由于顺铂引起的细胞凋亡,降低卵巢癌细胞对于顺铂的敏感性。Ku86对顺铂在卵巢癌药物敏感性中的这种影响可能是通过调节TOP1和COPS5发挥作用的。结论: TOP1和COPS5都是与DNA损伤修复有关并能影响铂类药物化疗敏感性的重要分子,下调Ku86会使TOP1的表达增高和COPS5的表达减少。卵巢癌中与铂类药物敏感性有关的生物标志物之间有相关性,靶向Ku86的治疗可能是提高卵巢癌药物敏感性的有效策略。

关键词: Ku86, 拓扑异构酶Ⅰ, COP9信号复合体的第5种成分, 顺铂, DNA损伤修复

Abstract: Background and purpose: Ovarian cancer is the disease with the third highest incidence and the highest lethality among gynecological malignancies. Ovarian cancer has a poor prognosis because most patients are in advanced stages at the time of diagnosis and susceptible to resistance to platinum-based chemotherapy. Abnormal DNA repair is an important cause of platinum resistance. Targeting interference with DNA damage repair molecules may be a new way to improve the sensitivity to platinum chemotherapy. Ku86 is a key molecule involved in the process of non-homologous end joining (NHEJ), which can effectively repair DNA double-strand break (DSB). Methods: Immunohistochemical staining and cellular immunofluorescence were used to detect the localization of Ku86 in ovarian cancer tissues and cells. Ku86 was downregulated by RNAi technology, apoptosis was detected by flow cytometry after treatment with cisplatin, and IC 50 was detected by cell counting kit-8 (CCK-8) method. Western blot was used to detect and analyze the relationship between Ku86 and topoisomerase Ⅰ(TOP1) and the fifth component of the COP9 signalosome (COPS5). Results: This study found that downregulating Ku86 reduced the apoptosis caused by cisplatin and decreased the sensitivity of ovarian cancer cells to cisplatin. This effect of Ku86 on cisplatin in ovarian cancer drug sensitivity might be mediated by the regulation of TOP1 and COPS5. Conclusion: TOP1 and COPS5 are both important molecules related to DNA damage repair and can affect platinum sensitivity in chemotherapy. Our study found that downregulating Ku86 increased the expression of TOP1 and decreased the expression of COPS5. This study preliminarily elucidated the relationship between biomarkers related to platinum drug sensitivity in ovarian cancer. Ku86-targeted therapy may be an effective strategy to improve drug sensitivity in ovarian cancer.

Key words: Ku86, Topoisomerase Ⅰ, The fifth component of the COP9 signalosome, Cisplatin, DNA damage repair