摘要
目的研究百合多糖1(LP1)对二甲双胍的抗乳腺癌细胞作用的影响。方法培养MCF-7乳腺癌细胞,加入(0.5、1.0)mg/m L LP1和(5、10、20、50)mmol/L的二甲双胍,MTT法检测LP1和二甲双胍对MCF-7细胞增殖的影响;异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双染色结合流式细胞术观察细胞周期和细胞凋亡;Western blot法检测Bcl-2、Bax、腺苷酸活化蛋白激酶(AMPK)和磷酸化腺苷酸活化蛋白激酶(p-AMPK)蛋白含量。结果 1 mg/m L LP1可以增强二甲双胍的抗增殖作用、细胞周期阻滞于G1期作用和诱导细胞凋亡作用。LP1处理增强二甲双胍对Bcl-2蛋白的下调和Bax蛋白的上调作用,但不影响二甲双胍对MAPK通路的激活作用。结论 LP1增强二甲双胍对MCF-7乳腺癌细胞的抗增殖作用,促进细胞凋亡,其机制与凋亡蛋白Bcl-2和Bax参与有关。
Objective To investigate the effect of metformin,alone or in combination with Lily polysaccharide 1（ LP1）,on cell viability and apoptosis in MCF-7 human breast cancer cells. Methods LP1（ 0. 5,1. 0 mg/m L） and metformin（ 5,10,20,50 mmol/L） were added into MCF-7 cell culture medium,followed by incubating for 72 hours in carbon dioxide incubators at 37℃. MCF-7 cell proliferation was determined using MTT assay; the apoptosis and cell cycle of MCF-7 cells were examined using annexinⅤ-FITC/PI double staining combined with flow cytometry; Western blotting was used to determine the content of Bcl-2,Bax,adenosine monophosphate-activated protein kinase（ AMPK） and phosphorated AMPK（ p-AMPK）proteins. Results Metformin-induced inhibition of MCF-7 cell proliferation was significantly enhanced when 1 mg/m L LP1 was added in. Compared with the control group and the metformin only group,more cells were arrested to G1 and the apoptosis rate was raised obviously in the metformin and LP1 combination group. LP1 promoted the downregulated expression of Bcl-2and the upregulated expression of Bax induced by metformin,but it didn＇t show any impact on the metformin-activated AMPK pathway. Conclusion LP1 enhances the proliferation-inhibitory and apoptosis-promoting effect of metformin on human breast carcinoma cells. The mechanism may be related with Bcl-2 downregulation and Bax upregulation.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第6期780-783,788,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
陕西省中医药管理局课题(15-ZY036)
陕西省自然科学基础研究计划(2016JM8047)
陕西省教育厅专项科研项目(2010JK805)
西安医学院学科建设经费基金(2016年)
西安医学院博士科研启动基金(2011DOC04)
作者简介
侯进（1973-），女，陕西西安人，副教授，博士Tel：029—86177562；E—mail：houandjin@sohu．com
通讯作者，弥曼，E-mail：mmpmm@263．net
