丹参酮ⅡA对乳腺癌细胞的影响及其分子机制

The influence of tanshinone Ⅱ A on breast cancer cells and its molecular mechanism

目的观察丹参酮ⅡA(TanⅡA)对乳腺癌细胞增殖和凋亡的影响,并探讨其分子机制。方法选取处于对数生长期的乳腺癌MCF-7细胞,设对照组(0μg/ml TanⅡA)和处理组(0.25、0.50、0.75μg/ml TanⅡA),处理MCF-7细胞48 h。MTT法检测各组MCF-7细胞的增殖抑制率,流式细胞术检测各组MCF-7细胞的凋亡率,Western Blot和RT-PCR分别检测各组MCF-7细胞中p53、Bcl2的蛋白和m RNA表达水平。结果与对照组比较,不同剂量TanⅡA处理组的MCF-7细胞增殖抑制率升高(P﹤0.05),且呈剂量-时间依赖性;随着TanⅡA作用剂量的增加,MCF-7细胞的凋亡率逐渐增大,均高于对照组(P﹤0.05)。与对照组比较,0.75μg/ml TanⅡA处理组的p53蛋白表达升高,Bcl2蛋白表达水平下降,差异有统计学意义(P﹤0.05);与对照组比较,0.75μg/ml TanⅡA处理组的p53 m RNA表达水平增加,Bcl2 m RNA表达水平降低,分别为对照组的4.88倍和0.42倍(P﹤0.05);0.75μg/ml TanⅡA处理组的p-AKT蛋白表达水平(0.866±0.015)低于对照组的p-AKT蛋白表达水平(1.000±0.020),差异有统计学意义(P﹤0.05)。结论 TanⅡA对乳腺癌MCF-7细胞具有抑制增殖和促进凋亡的作用,其凋亡作用与p53和Bcl2蛋白的表达有关,其抗凋亡机制与p-AKT信号通路有关。

Objective To observe the effect of tanshinone Ⅱ A（Tan Ⅱ A） on breast cancer cell proliferation and apoptosis, and to explore the underlying molecular mechanism. Method MCF-7 cells in logarithmic growth phase were selected and divided into control group（0 μg/ml Tan Ⅱ A） or study group（treated with 0.25, 0.50, 0.75 μg/ml Tan Ⅱ A）,which were treated with various concentrations of Tan Ⅱ A for 48 h. MTT assay was used to detect the inhibition rate of cell proliferation of MCF-7 cells in each group, and flow cytometry was utilized to determine the apoptosis rate, besides,Western Blot and RT-PCR were applied to detect the p53 and Bcl2 protein and m RNA expression level in each group. Re Result Compared with the control group, the inhibition rates of MCF-7 cells proliferation were increased（P0.05） in a time-and dose-dependent manner; with the increase of Tan Ⅱ A dose, the apoptosis rate of MCF-7 cells elevated gradually, which were higher than that of the control group（P0.05）. Compared with the control group, MCF-7 cells treated with0.75 μg/ml Tan Ⅱ A had increased expression of p53 and decreased expression of Bcl2 protein, with significant difference observed（P0.05）; besides, the expression of p53 m RNA were also increased, and the Bcl2 m RNA expression decreased in the MCF-7 cells after treatment with 0.75 μg/ml Tan Ⅱ A, respectively, and were 4.88 times and 0.42 times more than those in control group, respectively（P0.05）; while the expression level of p-AKT protein in MCF-7 cells treated with0.75 μg/ml Tan Ⅱ A（0.866 ± 0.015） was lower than that of control group at（1.000 ± 0.020）, and the difference was statistically significant（P0.05）. Conclusion Tan Ⅱ A can inhibit cell proliferation while promote apoptosis of breast cancer MCF-7 cells, and the effect is related to the expression of p53 and Bcl2 proteins, meanwhile, its anti-apoptotic mechanism is associated with p-AKT signaling pathway.