姜黄素对三阴乳腺癌MDA-MB-468细胞凋亡的作用

Effect of curcumin on apoptosis of MDA-MB-468 cells in triple-negative breast cancer

目的探讨姜黄素对人源性三阴乳腺癌MDA-MB-468细胞凋亡的影响,并从Akt信号通路阐述其可能机制。方法选取处于对数生长期的三阴乳腺癌MDA-MB-468细胞作为研究对象,给予不同浓度的姜黄素(35μmol/L、50μmol/L)分别处理12 h、24 h、48 h、72 h后,采用CCK8检测细胞的活性。将细胞分为对照组、DMSO组、姜黄素35μmol/L组、姜黄素50μmol/L组,干预48 h后,采用AnnexinV-FITC/PI双染法及流式细胞术检测细胞凋亡率,Western blot法检测细胞中凋亡相关蛋白(Akt、p-Akt、Bcl-2、Bax、Caspase-9、Caspase-3)的蛋白表达水平。结果姜黄素作用于MDA-MB-468细胞12 h、24 h、48 h后,50μmol/L姜黄素组的细胞抑制率均高于35μmol/L剂量组,在24 h细胞抑制率最高,差异均有统计学意义(P<0.05),但24 h后,细胞抑制率开始下降,至72 h时已低于5%;姜黄素处理48 h后,对照组、DMSO组、姜黄素35μmol/L组、姜黄素50μmol/L组细胞凋亡率分别为(31.29±0.51)%、(35.73±0.82)%、(40.22±1.64)%、(46.32±0.70)%,姜黄素干预组均高于对照组与DMSO组,且姜黄素50μmol/L组的细胞凋亡率高于姜黄素35μmol/L组,差异均有统计学意义(P<0.05);与对照组和DMSO组相比,两个姜黄素干预组p-Akt、Bcl-2的相对蛋白表达量均降低,而Bax、Caspase-9、Caspase-3的相对蛋白表达量则升高,差异均有统计学意义(P<0.05);其中姜黄素50μmol/L组p-Akt的相对蛋白表达量低于姜黄素35μmol/L组,而Bcl-2、Caspase-9的相对蛋白表达量则高于姜黄素35μmol/L组,差异均有统计学意义(P<0.05);但各组Akt的相对蛋白表达量之间比较差异无统计学意义(P>0.05)。结论姜黄素能够抑制三阴乳腺癌MDA-MB-468细胞增殖,且高浓度剂量组作用更强,可能与Akt信号通路的抑制有关。

Objective To investigate the effect of curcumin on apoptosis of MDA-MB-468 cells of human triple-negative breast cancer, and to explain its possible mechanism in the view of Akt signaling pathway.Methods MDA-MB-468 cells in the logarithmic growth phase were selected as the study subjects. Different concentrations of curcumin(35 μmol/L, 50 μmol/L) were administered for 12, 24, 48, and 72 hours, respectively. The activity of the cells was examined using CCK8. The cells were divided into the control group, DMSO group, curcumin 35 μmol/L group and curcumin 50 μ mol/L group. After 48 hours of intervention, apoptosis rate was detected by AnnexinV-FITC/PI double staining and flow cytometry. The protein expression levels of Akt, p-Akt, Bcl-2, Bax, Caspase-9 and Caspase-3 were detected by Western blot. Results When curcumin was applied to MDA-MB-468 cells for 12, 24, and48 hours, the cell inhibition rate of the 50 μmol/L curcumin group was higher than that of the 35 μmol/L group, and the cell inhibition rate was the highest at 24 hours significantly(P<0.05). However, after 24 hours, the cell inhibition rate began to decrease, and was less than 5% after 72 hours;after 48 hours of curcumin treatment, the apoptosis rates of the control group, DMSO group, curcumin 35 μ mol/L group and curcumin 50 μ mol/L group were(31.29 ± 0.51)%,(35.73 ±0.82)%,(40.22±1.64)%,(46.32±0.70)%, respectively. The curcumin intervention group was higher than the control and DMSO group, while the apoptosis rate of curcumin 50 μmol/L group was higher than that of curcumin 35 μmol/L group(P<0.05). Compared with the control group and the DMSO group, the relative protein expression levels of p-Akt and Bcl-2 were decreased in the two curcumin intervention groups, while that of Bax, Caspase-9 and Caspase-3 were increased(P<0.05). The relative protein expression of p-Akt in curcumin 50 μmol/L group was lower than that in curcumin 35 μmol/L group, while the relative protein expression of Bcl-2 and Caspase-9 was higher than those in curcumin 35 μmol/L group(P<0.05). However, there was no significant difference in the relative protein expression of Akt between the groups(P>0.05). Conclusion Curcumin can inhibit the proliferation of MDA-MB-468 cells in triple-negative breast cancer. The effect of high concentration dose is stronger, which may be related to the inhibition of Akt signaling pathway.