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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 pingyangmycin (PYM) induced senescence in human oral cancer cells. METHODS: Cell proliferation inhibition was detected by MTT assay and clonogenic assay. Aging-related β-galactoside The senescence-associatedβ-galactosidase (SA-β-gal) staining was used to observe the cellular senescence. The cell cycle distribution and intracellular reactive oxygen species (ROS) levels were measured by flow cytometry. The protein expression was detected by Western blotting . Results: The proliferation inhibitory activity of PYM on a variety of human cancer cells were stronger than that of bleomycin A2. Among them, KB cells are the most sensitive to PYM. PYM can also cause the aging of KB cells, which is related to cell arrest in G2 / M phase, intracellular ROS accumulation and the increase of p53 and p21 proteins. Conclusion: PYM causes cellular senescence and is also one of the mechanisms of its inhibition on KB cell proliferation.