姜黄素对人甲状腺癌B-CPAP细胞生物学行为的影响

摘要

Objective:To investigate the effect of curcumin on the growth and apoptosis of human thyroid carcino-ma B-CPAP cells and its possible mechanism.Methods:The human thyroid carcinoma B-CPAP cells were cul-tured in medium containing different concentrations (10 μmol/L,20 μmol/L,40 μmol/L)of curcumin for 12 h,24 h,36 h,48 h,72 h and 96 h.The effect of curcumin on the growth of B-CPAP cells was observed by MTT.The hu-man thyroid carcinoma B-CPAP cells were cultured in medium containing different concentrations of curcumin for 36 h,48 h,72 h and 96 h.The effects of curcumin in inducing apoptosis of B-CPAP cells were evaluated with Annexin V/PI staining,and the effect of curcumin on ERK pathway,Bax and Bcl-2 expression was detected by Western blot.Results:The growth inhibition rate and apoptosis rate of human thyroid carcinoma B-CPAP cells cultured in medium containing 10 μmol/L,20 μmol/L and 40 μmol/L of curcumin increased gradually with the prolongation of culture time (P＜0.05).With the increase of concentration,the growth inhibition rate and the apoptosis rate also increased gradually at the same time point (P＜0.05).After 72 h of culture,the expression levels of H-Ras,p-Raf-B,p-ERK1/2 and Bcl-2 proteins in B-CPAP cells were significantly lower,while the expression level of Bax protein was significantly higher than that in the blank control group (P＜0.05).Conclusion:Curcumin can inhibit the growth and induce apoptosis of human thyroid carcinoma B-CPAP cells.The mechanism may be related to inhibiting the ex-pression of H-Ras protein and ERK signaling pathway,up-regulating the expression of Bax and down regulating the expression of Bcl-2.%目的:探讨姜黄素对人甲状腺癌B-CPAP细胞生长、凋亡的影响及可能的作用机制.方法:采用含有不同浓度(10 μmol/L、20 μmol/L、40 μmol/L)姜黄素的培养基培养人甲状腺癌B-CPAP细胞12 h、24 h、36 h、48 h、72 h、96 h,应用MTT实验观察姜黄素对B-CPAP细胞生长的影响;采用含有不同浓度姜黄素的培养基培养人甲状腺癌B-CPAP细胞36 h、48 h、72 h、96 h,应用Annexin V/PI染色评价姜黄素诱导B-CPAP细胞凋亡的作用;应用蛋白印迹法检测姜黄素对ERK通路、Bax、Bcl-2表达的影响.结果:浓度为10 μmol/L、20 μmol/L、40 μmol/L的姜黄素培养甲状腺癌B-CPAP细胞,随着培养时间延长,各浓度下细胞生长抑制率、细胞凋亡率均逐渐增高(P＜0.05);且随着浓度增加,相同时间点的细胞生长抑制率、细胞凋亡率亦逐渐增高(P＜0.05).与空白组比较,姜黄素培养72 h后,B-CPAP细胞的H-Ras、p-Raf-B、p-ERK1/2、Bcl-2蛋白表达水平明显降低(P＜0.05),Bax蛋白表达水平水平明显升高(P＜0.05).结论:姜黄素能够抑制人甲状腺癌B-CPAP细胞生长,诱导细胞凋亡,其机制可能与抑制H-Ras蛋白、ERK信号通路蛋白表达及上调Bax表达、下调Bcl-2表达有关.