活性氧生成在卡铂诱导HN-3人喉鳞癌细胞凋亡中的作用  

Role of reactive oxygen series generation in the apoptosis of HN-3 human laryngeal carcinoma cells induced by carboplatin

作　　者：毛雯静 张焕康[1] 沈波[2] 何培杰[1] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻喉科,上海200031 [2]复旦大学放射医学研究所,上海200032

出　　处：《中国眼耳鼻喉科杂志》2015年第6期384-387,共4页Chinese Journal of Ophthalmology and Otorhinolaryngology

基　　金：国家自然科学基金面上项目(81172557);教育部留学回国人员科研启动基金(第45批)

摘　　要：目的观察卡铂(carboplatin)对HN-3人喉鳞癌细胞体外增殖的抑制作用、凋亡诱导,探讨活性氧生成在其中的作用。方法用卡铂以不同孵育浓度、不同孵育时间处理HN-3细胞,四甲基偶氮唑盐(MTT)法测定细胞活力,2',7'二氯氢化荧光素已二脂(H2DCFDA)染色测定活性氧生成,Hoechst/PI双重染色检测细胞凋亡/坏死情况,蛋白印迹法(Western blotting)测定多聚ADP-核糖聚合酶(PARP)表达。结果卡铂对体外培养的HN-3细胞有增殖抑制作用,呈剂量依赖性和时间依赖性;卡铂诱导HN-3细胞活性氧生成及凋亡呈时间与剂量依赖性;卡铂诱导氧化压力下PARP表达上调。结论卡铂可抑制人喉鳞癌HN-3细胞增殖,诱导HN-3细胞凋亡,其机制可能与卡铂诱导HN-3细胞活性氧生成及氧化压力下PARP表达上调有关。Objective To investigate the role of reactive oxygen series （ROS） in suppressing cell viability and inducing apoptosis of HN-3 human laryngeal squamous carcinoma cells induced by carboplatin. Methods The attached HN-3 cancer cells were treated with different concentrations of carboplatin for 24 h and 0.08 mg/mL carboplatin at indicated time points. Methyl thiazolyl tetrazolium（MTF） assay was used to detect cellular viability. H2DCFDA staining and Hoechst 33343/PI double staining were performed to monitor reactive oxygen series （ROS） generation and apoptosis/ necrosis, respectively. Western blotting technique was used to assess the expression of poly ADP-ribose polymerase （PARP）. Results Suppression of cellular viability on HN-3 cells induced by carboplatin was exhibited in a dose and time related pattern. Remarkable generation of ROS and apoptosis of HN-3 cells were initiated by carboplatin in dose and time related manner. Up-regulated expression of PARP under oxidative stress, induced by carboplatin, was partially inhibited by co-incubated with glutathione. Conclusions Generation of ROS and up-regulated PARP may play an important role in the suppression of cellular viability and apoptosis induction of HN-3 cells exerted by carboplatin.

关 键 词：喉鳞癌 卡铂 活性氧 凋亡 多聚ADP-核糖聚合酶 

分 类 号：R739.65[医药卫生—肿瘤]