平阳霉素诱导人口腔上皮癌KB细胞衰老的特征  

Characteristics of cellular senescence induced by pingyangmycin in human oral epithelial carcinoma KB cells

作　　者：桂冠[1] 王婷[2] 高瑞娟[2] 翟力新[3] 赵春燕[2] 王真[2] 李电东[2] 

机构地区：[1]南京医科大学第一附属医院,南京210029 [2]中国医学科学院医药生物技术研究所,北京100050 [3]南京大学医学院附属鼓楼医院,南京210008

出　　处：《中国新药杂志》2015年第18期2060-2064,2072,共6页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项(2009ZX09103-136;2013ZX09102064);国家自然科学基金(81001021)

摘　　要：目的:观察平阳霉素(pingyangmycin,PYM)诱导人口腔上皮癌细胞衰老的特征。方法:采用MTT[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide]法和克隆形成实验检测细胞增殖抑制作用;衰老相关β-半乳糖苷酶(senescence-associatedβ-galactosidase,SA-β-gal)染色法观察细胞衰老;流式细胞仪测定细胞周期分布和细胞内活性氧自由基(reactive oxygen species,ROS)水平;Western blotting检测蛋白表达水平。结果:PYM对多种人癌细胞的增殖抑制活性均强于博莱霉素A2。其中,KB细胞对PYM作用最敏感。PYM还可使KB细胞出现衰老特征,这与细胞阻滞于G2/M期,细胞内ROS积累以及衰老相关蛋白p53和p21水平增加有关。结论:PYM引起细胞衰老,也是其抑制KB细胞增殖的机制之一。Objective： To observe the characteristics of cellular senescence induced by pingyangmycin （PYM） in human oral epithelial carcinoma KB ceils. Methods： MTT reduction assay and colony-forming assay were used to detect the inhibitory effect on cell proliferation. Cellular senescence was observed using senescence-as- sociated 13-galactosidase staining. Flow cytometry was performed to measure the distribution of cell cycle and the level of intracellular reactive oxygen species （ROS）. Protein expression was analyzed by Western blotting. Re- sults： The anti-proliferative effect of PYM on various human carcinoma cells was stronger than that of bleomycin A2. Among the tested cell lines, the human oral epithelial carcinoma line, KB cells, was the most sensitive to PYM. PYM also induced the features of cellular senescence in KB cells, which was related to the arrest at G2/M phase, the accumulation of the intracellular ROS and the increased expression of the senescence-associated protein p53 and p21. Conclusion ： Cellular senescence induced by PYM is one of its important mechanisms in suppressing the proliferation of KB cells.

关 键 词：平阳霉素 博莱霉素A2 细胞增殖 细胞衰老 

分 类 号：R979.1[医药卫生—药品]