论文部分内容阅读
目的:研究川陈皮素对非小细胞肺癌NCI-H460细胞tubulin、Raf-1、Bcl-2表达的影响。方法:MTT法检测川陈皮素对NCI-H460细胞的生长抑制作用;用PI-Hoechst荧光双染法观察细胞凋亡形态;Western blot检测川陈皮素对NCI-H460细胞tubulin、Raf-1及Bcl-2蛋白表达的影响;体外微管蛋白聚合-解聚平衡体系研究川陈皮素对微管蛋白聚合的影响。结果:川陈皮素体外作用NCI-H460细胞24、48、72h后的IC50分别为23.5、11.3、3.81μmol/L;荧光双染色法显示经20μmol/L川陈皮素作用后细胞出现变形,染色质浓缩,产生凋亡小体;40μmol/L川陈皮素作用细胞3、6、12、24h后,未聚合的tubulin及Raf-1表达逐渐增加,Bcl-2表达呈下降趋势;10μmol/L川陈皮素在体外能破坏微管蛋白的动态平衡体系,抑制微管蛋白的聚合。结论:川陈皮素能抑制细胞中微管蛋白的聚合从而影响Raf-1及Bcl-2这一信号途径,这可能是其在体外抑制非小细胞肺癌NCI-H460细胞的生长且诱导凋亡机制。
Objective: To study the effects of nobiletin on the expression of tubulin, Raf-1 and Bcl-2 in NCI-H460 cell line. METHODS: MTT assay was used to determine the inhibitory effect of nobiletin on the growth of NCI-H460 cells. PI-Hoechst staining was used to observe the morphology of apoptosis. Western blot was used to detect tubulin, tubf, Raf-1 and Bcl in NCI-H460 cells. -2 Protein expression effects; in vitro tubulin polymerization-depolymerization equilibrium system Chuanxilae effect on tubulin polymerization. RESULTS: The IC50 values of nobiletin treated with NCI-H460 cells at 24, 48, and 72 h were 23.5, 11.3, and 3.81 μmol/L, respectively. The double staining showed that the cells were deformed and chromatin was treated with 20 μmol/L nobiletin. Concentration resulted in apoptotic bodies; after 3, 6, 12 and 24 h of treatment with 40 μmol/L nobiletin, the expression of unpolymerized tubulin and Raf-1 increased, and the expression of Bcl-2 decreased; 10 μmol/L Chuanci In vitro, it can destroy the dynamic equilibrium system of tubulin and inhibit the polymerization of tubulin. CONCLUSION: ChuanZeowin can inhibit the polymerization of tubulin in cells and thus influence the signaling pathways of Raf-1 and Bcl-2. This may be due to its inhibition of the growth of NCI-H460 cells in vitro and induction of apoptosis. .