雷公藤甲素经丝裂原活化蛋白激酶信号通路影响乳腺癌细胞增殖与凋亡的研究

Triptolide affects proliferation and apoptosis of breast cancer cells via mitogen-activated protein kinases signaling pathway

目的探讨雷公藤甲素经丝裂原活化蛋白激酶(MAPK)信号通路影响乳腺癌细胞增殖和凋亡的作用机制。方法分别用不同浓度(0,5,10,20,40和80nmol·L^-1)雷公藤甲素对MCF-7细胞进行干预。用CCK-8法检测细胞的增殖抑制率,用Western-blotting法检测雷公藤甲素对MCF-7细胞凋亡相关蛋白Bcl-2、Bax、Caspase-3、Caspase-9及MAPK信号通路相关蛋白的表达。结果干预48 h后,0,5,10,20,40和80 nmol·L^-1雷公藤甲素组的细胞抑制率分别为(100.00±0)%,(87.33±3.96)%,(61.67±4.09)%,(52.67±5.10)%,(37.67±2.98)%和(27.67±3.03)%,雷公藤甲素对MCF-7细胞增殖抑制作用与浓度呈正相关。干预48 h后,0,10,20和40 nmol·L^-1雷公藤甲素组的pJNK蛋白分别为(0.05±0.01),(0.12±0.04),(0.32±0.03)和(0.56±0.04),p-P38MAPK蛋白分别为(0.09±0.01),(0.23±0.03),(0.34±0.05)和(0.52±0.09),p-ERK蛋白分别为(0.46±0.07),(0.28±0.03),(0.18±0.06)和(0.09±0.05),Bcl-2蛋白分别为(1.48±0.23),(1.18±0.12),(0.80±0.05)和(0.32±0.06),Bax蛋白分别为(1.13±0.11),(1.27±0.18),(1.82±0.20)和(2.14±0.21),Caspase-3蛋白分别为(0.28±0.04),(1.04±0.08),(1.31±0.12)和(1.29±0.17),Caspase-9蛋白分别为(0.18±0.02),(0.44±0.07),(1.25±0.12)和(1.13±0.11),10,20和40nmol·L^-1雷公藤甲素组的上述指标与0 nmol·L^-1雷公藤甲素组比较,差异均有统计学意义(P<0.05或P<0.01)。结论雷公藤甲素可抑制人乳腺癌MCF-7细胞增殖,并通过下调Bcl-2蛋白表达,上调Bax、Caspase-3和Caspase-9蛋白表达,从而达到促进MCF-7细胞凋亡,其作用机制可能与MAPK信号通路有关。

Objective To investigate the mechanism of triptolide-induced mitogen-activated protein kinase(MAPK)signaling pathway in the proliferation and apoptosis of breast cancer cell.Methods MCF-7 cells were treated with different concentrations(0,5,10,20,40 and 80 nmol·L^-1)of triptolide.The proliferation inhibition rate of cells was detected by CCK-8 method.The apoptosis-related proteins Bcl-2,Bax,Caspase-3,Caspase-9 and MAPK signaling pathway-related proteins of MCF-7 cells were detected by Western-blotting method.Results After 48 h of intervention,the cell inhibition rates of the 0,5,10,20,40 and 80 nmol·L^-1 triptolide groups were(100.00±0)%,(87.33±3.96)%,(61.67±4.09)%,(52.67±5.10)%,(37.67±2.98)%and(27.67±3.03)%,the inhibition of proliferation of MCF-7 cells by triptolide was positively correlated with the concentration.After 48 h of intervention,the p-JNK protein of 0,10,20 and 40 nmol·L^-1 triptolide groups were(0.05±0.01),(0.12±0.04),(0.32±0.03)and(0.56±0.04),p-P38MAPK protein were(0.09±0.01),(0.23±0.03),(0.34±0.05)and(0.52±0.09),p-ERK protein were(0.46±0.07),(0.28±0.03),(0.18±0.06)and(0.09±0.05),Bcl-2 protein were(1.48±0.23),(1.18±0.12),(0.80±0.05)and(0.32±0.06),Bax protein were(1.13±0.11),(1.27±0.18),(1.82±0.20)and(2.14±0.21),Caspase-3 protein were(0.28±0.04),(1.04±0.08),(1.31±0.12)and(1.29±0.17),Caspase-9 protein were(0.18±0.02),(0.44±0.07),(1.25±0.12)and(1.13±0.11),the differences were statistically significant between 10,20,40 nmol·L^-1 triptolide groups and 0 nmol·L^-1 triptolide group(P<0.05 or P<0.01).Conclusion Triptolide can inhibit the proliferation of human breast cancer MCF-7 cells and promote the apoptosis of MCF-7 cells by down-regulating the expression of Bcl-2 protein and up-regulating the expression of Bax,Caspase-3 and Caspase-9.Its mechanism may be related to MAPK signaling pathway.