摘要
背景与目的:极光激酶A(Aurora A kinase,Aurora A)属于丝/苏氨酸蛋白激酶家族中的一员,可促进卵巢癌的化疗抵抗。活性氧(reactive oxygen species,ROS)由机体正常代谢产生,参与细胞信号转导过程。肿瘤细胞由于代谢旺盛及细胞内氧化还原体系的失衡,ROS水平高于正常细胞。并且耐药细胞株中ROS水平降低,提示ROS在肿瘤耐药中发挥作用。该研究旨在探讨ROS在Aurora A介导顺铂化疗耐药中的作用。方法:采用二氯二氢荧光素-乙酰乙酸酯(2’,7’-dichlorofluorescin diacetate,DCFH-DA)法检测人卵巢癌细胞株A2780和顺铂耐药株A2780/CDDP中ROS的水平,采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法检测顺铂的半数抑制浓度(50%inhibitory concentration,IC50);构建Aurora A过表达和沉默细胞株,检测细胞内ROS水平;加入ROS清除剂N-乙酰-L-半胱氨酸(N-acetyl-L-cysteine,NAC),检测顺铂的IC50;采用蛋白[质]印迹法(Western blot)检测相关信号通路,探讨可能的分子机制。结果:顺铂耐药株中ROS水平低于对照细胞株。加入NAC降低细胞内ROS可增强细胞对顺铂的耐药性。Aurora A过表达,细胞内ROS水平下降,可增强细胞对顺铂的耐药;而Aurora A沉默后,细胞内ROS升高,则可提高细胞对顺铂的敏感性。Aurora A过表达,促进AMP活化蛋白激酶(AMP-activated protein kinase,AMPK)的磷酸化及糖酵解。结论:Aurora A可能通过促进AMPK磷酸化及糖酵解过程,降低细胞内ROS水平,从而增强卵巢癌细胞对顺铂的耐药性。
Background and purpose： Aurora A kinase（Aurora A）, a member of serine/threonine kinase family, contributes to chemo-resistance in ovarian cancer. Reactive oxygen species（ROS）, generated during normal metabolism in aerobic organisms, are involved in multiple signaling pathways. Cancer cells possess higher ROS levels than normal cells due to the increased metabolism and cellular redox imbalance. Moreover, ROS levels are lower in chemo-resistant cells, suggesting the crucial role of ROS in chemo-resistance. However, the role of ROS in Aurora A-mediated chemo-resistance is largely unexplored. Methods： ROS levels of A2780 and A2780/CDDP were examined using 2＇, 7＇-dichlorofluorescin diacetate（DCFH-DA）, and 50% inhibitory concentration（IC_（50）） values of cisplatin were measured by methyl thiazolyl tetrazolium（MTT）. Establishment of stable ovarian cancer cell lines harboring Aurora A c DNA or sh RNA were conducted by Lentil-virus infection and subsequent drug screening. ROS levels and IC_（50） values of cisplatin were detected in these established cells. IC_（50） values of cisplatin were also measured in combination with N-acetyl-L-cysteine（NAC）. To explore the potential mechanism, cells were collected and subjected to Western blot. Results： Chemo-resistant cells exhibited lower ROS levels than control cells. Reduction of ROS levels by NAC promoted chemo-resistance. Overexpression of Aurora A inhibited ROS and conferred chemo-resistance. Knockdown of Aurora A elevated the ROS levels and promoted chemo-sensitivity. Aurora A promoted the phosphorylation of AMPactivated protein kinase（AMPK） and glycolysis. Conclusion： Aurora A promotes chemo-resistance by decreasing the cellular ROS levels. AMPK phosphorylation and glycolysis are involved in Aurora A-mediated ROS regulation.
作者
杨丽娜
杨宇琦
杨恭
陈亚萍
YANG Lina;YANG Yuqi;YANG Gong;CHEN Yaping(Department of Obstetrics and Gynecology, the Fifth People＇s Hospital of Shanghai, Fudan University, Shanghai 200240, China;Cancer Institute, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China;Central Laboratory, the Fifth People＇s Hospital of Shanghai, Fudan University, Shanghai 200240, China)
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2018年第3期184-190,共7页
China Oncology
基金
国家自然科学基金(81602259)
关键词
活性氧
极光激酶A
化疗抵抗
Reactive oxygen species
Aurora A kinase
Chemo-resistance
