摘要
目的:研究益气小复方在乳腺癌他莫昔芬(TAM)耐药中的作用及对胞内磷脂酞肌醇激酶(PI3K)/蛋白激酶B(AKT)通路的影响。方法:建立乳腺癌TAM耐药细胞(MCF-7/TAM)模型,细胞形态观察及MTT法测定细胞抑制率;不同浓度益气小复方处理MCF-7/TAM细胞,MTT法测定增殖抑制率,流式细胞仪测定凋亡率,蛋白印迹法(WB)测定细胞bax、bak、Bcl-2、PI3K、AKT、p-AKT、P-gp、PTEN蛋白表达水平。结果:与MCF-7细胞相比,MCF-7/TAM细胞形态不规则,细胞变大,触角延伸,细胞呈簇状生长,同浓度TAM处理后,MCF-7/TAM细胞抑制率明显降低,IC50值显著升高(P<0.05)。进一步研究益气小复方对MCF-7/TAM细胞增殖抑制发现,与MCF-7细胞相比,同浓度益气小复方处理后,MCF-7/TAM细胞细胞抑制率、IC50值无差异(P>0.05);选取40mg/ml、80mg/ml、160mg/ml进行后续实验,与MCF-7/TAM细胞组相比,低、中、高剂量益气小复方处理MCF-7/TAM细胞48h后,细胞凋亡率显著升高(P<0.05);与MCF-7细胞组相比,MCF-7/TAM细胞组PI3K、P-gp、Bcl-2蛋白表达水平、AKT磷酸化水平显著升高(P<0.05),PTEN蛋白表达水平、bax、bak显著降低(P<0.05);与MCF-7/TAM细胞组相比,低、中、高剂量益气小复方组PI3K、P-gp、Bcl-2蛋白表达水平、AKT磷酸化水平显著降低(P<0.05),PTEN蛋白表达水平、bax、bak显著升高(P<0.05),呈剂量依赖性。结论:益气小复方可能通过抑制PI3K/AKT通路活化,逆转乳腺癌他莫昔芬耐药。
Objective:To study the effect of Yiqi Xiaofu Prescription on tamoxifen(TAM)resistance in breast cancer and its influence on intracellular phospholipid phthalocyanine kinase(PI3K)/protein kinase B(AKT)pathway.Methods:The model of TAM-resistant breast cancer cells(MCF-7/TAM)was established,cell morphology and MTT assay were used to determine the inhibition rate.MCF-7/TAM cells were treated with different concentrations of Yiqi Xiaofu Prescription,the inhibition rate of proliferation was measured by MTT,the apoptotic rate was measured by flow cytometry,and the expression levels of bax,bak,Bcl-2,PI3K,AKT,p-AKT,P-gp and PTEN were determined by Western blotting(WB).Results:Compared with MCF-7 cells,MCF-7/TAM cells had irregular morphology,enlarged cells,extended antennae and clustered growth.After treated with TAM at the same concentration,the inhibition rate of MCF-7/TAM cells decreased significantly and the IC50 value increased significantly(P<0.05).Further study on the inhibition of Yiqi Xiaofu Prescription on the proliferation of MCF-7/TAM cells showed that compared with MCF-7 cells,after treatment with the same concentration of Yiqi Xiaofu Prescription,there was no difference in the inhibition rate and IC50 value of MCF-7/TAM cells(P>0.05);40 mg/ml,80 mg/ml and 160 mg/ml were selected for subsequent experiments,compared with the MCF-7/TAM cell group,the apoptotic rate of MCF-7/TAM cells increased significantly at 48 hours after treating with low,medium and high dose of Yiqi Xiaofang Prescription(P<0.05).Compared with the MCF-7 cell group,the expression levels of PI3K,P-gp,Bcl-2 protein and the level of AKT phosphorylation in the MCF-7/TAM cell group increased significantly(P<0.05),while the expression level of PTEN protein,Bax and Bak decreased significantly(P<0.05).Compared with the MCF-7/TAM cell group,the levels of PI3K,P-gp,Bcl-2 protein expression and the level of AKT phosphorylation in low,medium and high dose Yiqi Xiaofang groups decreased significantly(P<0.05),while the level of PTEN protein expression,Bax
作者
魏鑫
石朋飞
刘勇军
WEI Xin;SHI Pengfei;LIU Yongjun(Department of Thyroid and Breast Surgery,the Central Hospital of Wuhan,Tongji Medical College of Huazhong University of Science and Technology(Wuhan Hubei 430000,China)
出处
《四川中医》
2020年第7期56-61,共6页
Journal of Sichuan of Traditional Chinese Medicine
基金
湖北省科技计划项目(第二批)(编号:2019CFB124)。
关键词
益气小复方
胞内磷脂酞肌醇激酶
蛋白激酶B
乳腺癌
他莫昔芬
Yiqi Xiaofu Prescription
Intracellular phosphatidylinositol kinase
Protein kinase B
Breast cancer
Tamoxifen
