海绵来源的smenospongine诱导乳腺癌MCF7细胞凋亡机制研究

Effect of marine sponge-derived smenospongine on apoptosis in breast cancer MCF7 cells

目的研究海绵Spongia pertusa Esper来源的smenospongine(Sme)诱导乳腺癌细胞MCF7凋亡的作用机制。方法使用CCK-8法检测Sme对MCF7细胞活力的影响;DAPI染色检测凋亡细胞的细胞核形态;使用流式细胞术检测细胞凋亡率和线粒体膜电位;Western blotting检测Sme对Bax、Bcl2、细胞色素C(cytochorome C)、p-p38和p38蛋白水平表达的影响。结果 CCK-8法检测结果表明,Sme抑制MCF7增殖,IC50值为(16.46±0.88)μmol/L;DAPI染色结果和Annexin VFITC/PI染色结果显示Sme诱导细胞凋亡。随着加药浓度的增加,细胞凋亡率从4.18%逐渐升至21.49%;流式细胞仪检测结果表明Sme引发MCF7细胞内线粒体膜电位下降;Western blotting结果显示Sme激活了内源性凋亡途径和p38丝裂源活化蛋白激酶(MAPK)信号通路。结论 Sme可能通过激活p38 MAPK通路,诱导细胞内源性凋亡发挥抗乳腺癌作用。

Objective To investigate the potential effect of marine sponge-derived smenospongine（Sme）on apoptosis in breast cancer MCF7 cells.Methods The CCK-8 method was used to detect growth-inhibitory effect of Sme against MCF7 cells.Flow cytometry was used to evaluate the mitochondrial membrane potential and apoptosis.Changes in nuclear morphology of apoptotic cells were assessed by DAPI staining.Western blotting was used to detect the expression of Bax,Bcl2,cytochorome C,p38 and p-p38.Results Sme exhibited suppressive effect on the proliferation of MCF7 cells,with IC50 value of（16.46±0.88）μmol/L.DAPI staining and Annexin V-FITC/PI double staining revealed that Sme significantly induced apoptosis.The apoptosis rate was increased rapidly from 4.18% to 21.49% with the raise of Sme concentration.Further study showed that mitochondrial membrane potential decreased after Sme incubation.Western blotting analysis displayed that the expression of Bax was increased and the expression of Bcl2 was decreased,which resulted in the release of cytochorome C.Meanwhile,the phosphorylated level of p38 was significantly elevated.Conclusion Sme inhibited the proliferation of MCF7 cells and might activate intrinsic apoptosis through p38 MAPK pathway.