›› 2012, Vol. 24 ›› Issue (1): 30-34.doi: 10.3969/j.issn.1004-616x.2012.01.007

• 论著 • 上一篇    下一篇

NOX家族在X射线诱导PC-3细胞损伤中的作用

韩晓燕,刘 箐,高丽萍,马建秀,黄超杰,张 红   

  1. 1. 兰州大学基础医学院生物化学与分子生物学研究所,兰州 730000;2. 中国科学院近代物理研究所重离子辐照生物医学研究中心,兰州 730000;3. 上海理工大学医疗器械与食品学院,上海 200093;4. 西北民族大学医学院机能教研室,兰州 730030;5. 甘肃省出入境检验检疫局,兰州 730010
  • 收稿日期:2011-11-03 修回日期:2011-11-30 出版日期:2012-01-30 发布日期:2012-01-30
  • 通讯作者: 刘箐

Role of NOX family in PC-3 cell damage induced by X-ray irradiation

HAN Xiao-yan,LIU Qing,GAO Li-ping,MA Jian-xiu,HUANG Chao-jie,ZHANG Hong   

  1. 1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000;
  • Received:2011-11-03 Revised:2011-11-30 Online:2012-01-30 Published:2012-01-30
  • Contact: LIU Qing

摘要: 目的: 研究NOX家族在X射线诱导人雄激素非依赖性前列腺癌PC-3细胞损伤中的作用,寻找放疗增敏的潜在靶点。方法:采用噻唑蓝(MTT)比色法检测0、1、2、4和12 Gy X射线照射后24、48和96 h PC-3细胞存活率;采用DCFH-DA法检测0、1和4 Gy X射线照射后15、30、60和120 min 时PC-3细胞中ROS的生成量;采用Western blot方法检测0、1和4 Gy X射线照射后PC-3细胞中NOX1~NOX5 5个亚型蛋白的表达情况。结果:1、2、4和12 Gy X射线照射PC-3细胞后96 h,与未照射组比较,PC-3细胞的存活率明显下降 (P<0.05)。1和4 Gy X射线照射PC-3细胞60 min后,细胞内ROS水平最高,NOX抑制剂DPI及抗氧化剂N- 乙酰半胱氨酸(NAC)预处理可以减少ROS的生成。1和4 Gy X射线照射后PC-3细胞中NOX1、NOX2和NOX5蛋白表达显著增加。结论:X射线诱导NOX1、NOX2和NOX5蛋白过表达,细胞内产生过量的ROS是X射线诱导PC-3细胞损伤的机制之一。

关键词: NOX, 活性氧, 人前列腺癌细胞PC-3, X射线

Abstract: OBJECTIVE: To study the role of NOX (NADPH oxidase) in X-ray- induced damage of human androgen-independent prostate cancer PC-3 cells damage,search for potential targets for radiation sensitization. METHODS:The viability of human androgen-independent prostate cancer PC-3 cells induced by 0,1,2,4 and 12 Gy of X rays after 24,48 and 96 h was detected by the MTT assay. The level of ROS after X-ray irradiation for 15,30,60 and 120 min ,and the expression of NOX1-5 protein in PC-3 cells induced by 0,1 and 4 Gy of X rays was analyzed by using DCFH-DA as a probe and by Western blot,respectively. RESULTS:Compared with non-irradiated,the viability of human prostate cancer PC-3 cells induced by 1,2,4 and 12 Gy of X rays was significantly decreased (P<0.05) . The level of ROS reached a maximum at 60 min after 1 and 4 Gy of X-ray irradiation. NOX inhibitor DPI and antioxidant NAC pretreatment could reduce the generation of ROS. Western blotting showed the expressions of NOX1,NOX2 and NOX5 were increased after irradiation. CONCLUSION:X-ray-induced the homologs NOX1,NOX2 and NOX5 of the catalytic subunit gp91phox of NADPH oxidase over-expression,resulting in excessive intracellular ROS which is a new mechanism of X-ray-induced damage of prostate cancer cells.

Key words: NADPH oxidase, reactive oxygen species, human prostate cancer PC-3 cells, X-ray