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目的探讨姜黄素对髓母细胞瘤(medulloblastoma,MB)DAOY细胞增殖的影响及其机制。方法用20、40、60、80、100μmol/L的姜黄素(curcumin)处理DAOY细胞24、48和72 h,采用MTT法检测姜黄素对细胞增殖活性的影响;用30μmol/L姜黄素处理DAOY细胞48 h,设未处理细胞为对照组,免疫细胞化学法检测姜黄素对细胞中β-catenin蛋白表达的影响,Western blot法检测β-catenin及cylinD1蛋白的表达水平。结果不同浓度及作用不同时间姜黄素均可明显抑制DAOY细胞的增殖(P<0.05),且在姜黄素浓度≤60μmol/L时,呈时间、剂量依赖性。对照组细胞中,β-catenin蛋白在胞浆和胞核中均有表达,且以胞核为主,经30μmol/L姜黄素处理48 h后,胞核蛋白β-catenin及总蛋白cyclinD1的表达均明显低于对照组(P<0.05),胞浆蛋白β-catenin的表达无明显降低(P>0.05)。结论姜黄素可通过抑制β-catenin的表达和核转位,阻断Wnt/β-catenin信号通路转导,进而抑制其下游靶基因cyclinD1的表达,从而抑制MB的增殖。
Objective To investigate the effect of curcumin on the proliferation of medulloblastoma (MB) DAOY cells and its mechanism. Methods DAOY cells were treated with curcumin (20, 40, 60, 80, 100μmol / L) for 24,48 and 72 h, respectively. The effect of curcumin on the proliferation of cells was detected by MTT assay. DAOY The cells were treated with curcumin for 48 h. The untreated cells were used as the control group. The expression of β-catenin protein was detected by immunocytochemistry. The expression of β-catenin and cylinD1 protein was detected by Western blot. Results Curcumin significantly inhibited the proliferation of DAOY cells at different concentrations and times (P <0.05), and showed a time-and dose-dependent manner when curcumin concentration was ≤60 μmol / L. In the control group, β-catenin protein was expressed in the cytoplasm and nucleus, mainly in the nucleus. After treated with 30μmol / L curcumin for 48 h, the expression of β-catenin and cyclinD1 (P <0.05). The expression of β-catenin in cytoplasm was not significantly reduced (P> 0.05). Conclusion Curcumin can inhibit the proliferation of MB by inhibiting the expression and nuclear translocation of β-catenin, blocking the transduction of Wnt / β-catenin signaling pathway and inhibiting the expression of cyclinD1.