大戟大枣汤含药血清通过PI3k/Akt通路对乳腺癌细胞凋亡的影响

Effects of Decoction of Euphorbia fischeriana Steud.and Ziziphus jujuba Mill.medicated serum on apoptosis of breast cancer cells via pI3k/Akt pathway

目的探讨大戟大枣汤含药血清通过PI3k/Akt通路对乳腺癌细胞凋亡的影响。方法制备大戟大枣汤高、中、低剂量组及阴性组的含药血清,作用于体外培养的MCF-7细胞。吖啶橙-溴乙锭染色观察大戟大枣汤含药血清作用对细胞形态学的变化,Hoechst333342/PI染色分析细胞凋亡情况,免疫荧光法检测PI3k/Akt通路相关蛋白的表达,Western blot检测大戟大枣汤含药血清联合LY294002对PI3k/Akt通路相关蛋白的表达影响。结果荧光显微镜下可见大戟大枣汤含药血清作用后细胞贴壁能力下降、形态变得圆润、数量减少。与阴性对照组比较,大戟大枣汤含药血清组细胞凋亡增高(P<0.05,P<0.01),PI3k、p-Akt、p-FoxO3a蛋白表达降低(P<0.05,P<0.01);大戟大枣汤含药血清联合LY294002作用后,细胞中p-Akt、p-FoxO3a蛋白表达较单独使用大戟大枣汤含药血清更低。结论大戟大枣汤含药血清可通过抑制PI3k/Akt信号通路调控MCF-7细胞凋亡。

AIM To explore the effects of serum medicated with Decoction of Euphorbia fischeriana Steud.and Ziziphus jujuba Mill.(DEFSJ)on breast cancer cells apoptosis via pI3k/Akt pathway.METHODS The highdose,medium-dose,low-dose and negative DEFSJ medicated serum were prepared and applied to cultured MCF-7 cells in vitro.The effect of DEFSJ serum on cell morphology was observed using acridine orange-ethidium bromide(AO-EB)staining.The apoptosis of MCF-7 cells was analyzed with Hoechst333342/PI staining.The expression of associated PI3k/Akt pathway proteins was detected by immunofluorescence.Western blot assay was used to evaluate the influence of combinative use of DEFSJ medicated serum and LY294002 on the expression of PI3k/Akt pathway related proteins.RESULTS The cells exposed to DEFSJ medicated serum showed decreased cell adhesion ability,round shape,and less counts under fluorescence microscope observation.In contrast to the negative control group,DEFSJ medicated serum groups had higher apoptosis(P<0.05,P<0.01),and decreased expression of PI3k,p-Akt,and p-FoxO3a(P<0.05,P<0.01).The combinative use of DEFSJ medicated serum and LY294002 brought forth lower expression of p-Akt and p-FoxO3a protein in cells than that of the pure use of DEFSJ medicated serum.CONCLUSION DEFSJ medicated serum can regulate the apoptosis of MCF-7 cells by inhibiting the PI3k/Akt signaling pathway.