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鉴于细胞衰老是抑制肿瘤的重要步骤,考察博宁霉素引起人肿瘤细胞衰老的特征。采用MTT法和克隆形成率实验检测细胞增殖抑制作用;衰老相关的β-半乳糖苷酶染色检测细胞衰老;流式细胞仪测定细胞周期分布以及细胞内活性氧自由基水平;Western blotting检测蛋白的表达水平。结果表明,博宁霉素对人口腔上皮癌KB细胞的作用,明显强于人非小细胞肺癌A549细胞。与阳性对照药多柔比星的作用相似,博宁霉素可引起这两种细胞均出现衰老特征,这与细胞阻滞在G2/M期、细胞内活性氧自由基增加有关。0.1μmol·L-1博宁霉素可以诱导KB细胞衰老,检测到衰老标志蛋白P21的表达明显增加;而高浓度的博宁霉素激活细胞凋亡通路。本研究结果表明:博宁霉素引起细胞衰老,也是其抑制肿瘤细胞增殖的机制之一。
Since cell senescence is an important step in tumor suppression, we examined the characteristics of botulinum-derived senescence in human tumor cells. Cell proliferation inhibition was detected by MTT assay and clonogenic rate assay. Aging-related β-galactosidase staining was used to detect cell senescence. Flow cytometry was used to determine cell cycle distribution and intracellular reactive oxygen free radicals. Western blotting was used to detect protein The expression level. The results showed that the effect of Boningomycin on human oral epithelial cancer KB cells was significantly stronger than that of human non-small cell lung cancer A549 cells. Similar to the doxorubicin, a positive control, Boningomycin caused senescence in both cells, which was associated with an increase in reactive oxygen species (ROS) in G2 / M phase cells. At the concentration of 0.1μmol·L-1, bleomycin induced the senescence of KB cells, and the expression of P21, a marker of aging, was significantly increased. However, the high concentration of debinomycin activated the apoptotic pathway. The results of this study show that: Bonimycin causes cellular senescence and is also one of the mechanisms that inhibits the proliferation of tumor cells.