姜黄素抑制Wnt/β-catenin信号通路诱导乳腺癌细胞凋亡的机制研究

Study on the mechanism of curcumin inhibiting the apoptosis of breast cancer cells induced by Wnt / β- catenin signaling pathway

目的研究姜黄素对人乳腺癌细胞株BT549细胞凋亡的诱导作用及其机制。方法取对数生长期BT549细胞,制成单细胞悬液,计数并接种于96孔培养板,每孔约5 000个细胞,24 h后加浓度分别为0,10,20,40,80μmol/L的姜黄素,培养12,24,48 h后采用MTT实验检测BT549细胞增殖抑制率;分别用0,10,20,40,80μmol/L姜黄素处理细胞,48h后采用TUNEL染色法检测细胞凋亡指数;分别用0,10,20,40,80μmol/L姜黄素处理细胞,48 h后采用Western blot实验检测细胞增殖、凋亡相关蛋白的表达量。结果 MTT结果显示,随培养时间延长,姜黄素0,20,40,60,80μmol/L组BT549细胞增殖抑制率逐渐增高(P均<0.05),且同时间点内姜黄素0,20,40,60,80μmol/L组细胞增殖抑制率随剂量增加逐渐增高(P均<0.05)。TUNEL染色结果显示,姜黄素0,20,40,60,80μmol/L组BT549细胞凋亡指数随剂量增加逐渐增加(P均<0.05)。Western blot实验结果显示,随着姜黄素浓度的增加,总蛋白中β-catenin的表达不受影响(P>0.05),而细胞核中β-catenin的表达逐渐减少(P<0.05)。结论姜黄素通过Wnt/β-catenin信号传导通路抑制乳腺癌细胞株BT549细胞的增殖和诱导其凋亡,并呈现时间、剂量依赖性。

Objective It is to study the inducing effect of curcumin on apoptosis of human breast cancer cell line BT549 and its mechanism. Methods Logarithmic growth phase bt549 cells were counted and seeded on 96 hole culture plate with about 5 000 cells in each hole. After 24 h,0,10,20,40,80 μmol / L curcumin were added to culture cells,and the proliferation inhibition rate of BT549 cells was detected by MTT assay after 12,24 and 48 h culture. The cells were treated with 0,10,20,40,80 μmol / L curcumin respectively,and the apoptosis index was detected by TUNEL staining after 48 hours. The cells were treated with 0,10,20,40,80 μmol / L curcumin respectively,and the expression of cell proliferation and apoptosis related protein was detected by Western blot assay after 48 h. Results The MTT colorimetric assay results showed that at 12 h,24 h,48 h,the BT549 cells proliferation inhibition rate gradually increased in 0 μmol / L,20 μmol / L,40 μmol / L,60 μmol / L,80μmol /L curcumin group（ P 0. 05）. The results of TUNEL staining showed that the apoptotic index of BT549 cells increased gradually in the 0 μmol / L,20 μmol / L,40 μmol / L,60 μmol / L,80 μmol / L curcumin group（ P 0. 05）. Blot Western showed that with the increase of the concentration of curcumin,the expression of β- catenin in the total protein was not affected（ P 0. 05）,while the expression of β- catenin in the nucleus gradually decreased（ P 0. 05）. Conclusion Curcumin can inhibit the proliferation and induce the apoptosis of breast cancer cell line BT549 cells by β- catenin beta Wnt / signaling pathway,and present time and dose dependence.