迷迭香酸通过MAPK/ERK信号通路对结肠癌细胞增殖及凋亡的影响

Effects of Rosmarinic Acid on Proliferation and Apoptosis of Colon Cancer Cells via MAPK/ERK Signaling Pathway

[目的]探讨迷迭香酸通过丝裂原激活的蛋白激酶/胞外信号调节的蛋白激酶（MAPK/ERK）信号通路对结肠癌细胞增殖凋亡的影响。[方法 ]取对数生长期LS174T细胞,加入迷迭香酸（0、25、50和100μmol/L）处理24、48和72h后,CCK-8法检测迷迭香酸对细胞增殖活性的影响。以100μmol/L迷迭香酸处理细胞,24、48和72h后,采用流式细胞仪检测迷迭香酸对细胞周期和细胞凋亡率的影响;采用蛋白质印迹法检测细胞中Bax、Bcl-2和Cleaved Caspase-3蛋白的表达及ERK1/2磷酸化水平。[结果 ]迷迭香酸对LS174T细胞呈时间浓度依赖性抑制细胞增殖;100μmol/L迷迭香酸处理LS174T细胞不同时间后,与对照组相比,迷迭香酸组细胞中S期和G2/M期细胞所占比例明显降低,G0/G1期比例显著升高,但组间时间效应不显著（P〉0.05）;而Bcl-2蛋白表达、ERK1/2磷酸化水平明显降低,细胞凋亡率、Bax和Cleaved Caspase-3蛋白表达量显著增强,均表现出明显的时效关系（P〈0.05）。[结论 ]迷迭香酸能够抑制结肠癌LS174T细胞增殖,诱导细胞凋亡,其作用机制可能与抑制MAPK/ERK信号通路有关。

[Purpose] To investigate the effect of rosmarinic acid on the proliferation and apoptosis of colon cancer cells and its relation to MAPK/ERK signaling pathway. [Methods] The cultured human colorectal cancer LS174 T cells in logarithmic growth phase were treated with rosmarinic acid（0,25,50 and 100μmol/L）for 24,48 and 72 h,respectively. CCK-8 assay was used to detect the effect of rosmarinic acid on cell proliferation;flow cytometry was used to detect the effects of rosmarinic acid on cell cycle and cell apoptosis rate after LS174 T cells were treated with 100μmol/L rosmarinic acid for 24,48 and 72 h;the expressions of Bax,Bcl-2 and cleaved Caspase-3 proteins and phosphorylation level of ERK1/2 were detected by immunoblotting. [Results] Rosmarinic acid inhibited the proliferation of LS174 T cells in a time and dose-dependent manner. Compared with the control group,the proportion of S phase and G2/M phase cells in rosmarinic acid treatment group decreased and that of G0/G1 phase increased significantly,but the time effect between the groups was not significant（P〈0.05）. However,the expression of Bcl-2 protein and the level of ERK1/2 phosphorylation were significantly decreased,and the apoptosis rate,the expression of Bax and cleaved Caspase-3 protein were significantly increased in a time-dependent manner（P〈0.05）.[Conclusion] Rosmarinic acid can inhibit the proliferation of human colorectal cancer LS174 T cells and induce apoptosis,which may be related to the inhibition of MAPK/ERK signaling pathway.