摘要
目的:研究山慈菇是否通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路影响乳腺癌MDA-MB-231细胞的增殖和凋亡。方法:将MDA-MB-231细胞分为对照组(C组)、山慈菇组(CAM组)、胰岛素生长因子1组(IGF-1组)、山慈菇+胰岛素生长因子1组(CAM+IGF-1组)。C组不加药物处理,CAM组加入10 mg/ml山慈菇处理,IGF-1组加入100μg/L IGF-1处理,CAM+IGF-1组加入10 mg/ml山慈菇和100μg/L IGF-1共处理。噻唑蓝(MTT)测定细胞增殖,流式细胞仪和Hoechst 33258染色测定细胞凋亡,Western blot法测定细胞活化的半胱天冬氨酸3(Cleaved-Caspase-3)、Bcl-2相关X蛋白(Bax)、B-细胞淋巴瘤因子(Bcl-2)、PI3K、磷酸化PI3K(p-PI3K)、Akt、p-Akt蛋白水平。结果:山慈菇抑制MDA-MB-231细胞增殖呈剂量和时间依赖性。与C组比较,CAM组MDA-MB-231细胞OD值降低(P<0.05),细胞凋亡率升高(P<0.05),Cleaved-Caspase-3、Bax蛋白水平升高(P<0.05),Bcl-2、p-PI3K、p-Akt蛋白水平降低(P<0.05);与CAM组比较,CAM+IGF-1组MDA-MB-231细胞OD值升高(P<0.05),细胞凋亡率降低(P<0.05),Cleaved-Caspase-3、Bax蛋白水平降低(P<0.05),Bcl-2、p-PI3K、p-Akt蛋白水平升高(P<0.05)。结论:山慈菇抑制乳腺癌细胞增殖、诱导乳腺癌细胞凋亡,其机制可能与山慈菇抑制PI3K/Akt信号通路活化有关。
Objective:To investigate the affect of cremastra appendiculata makino on proliferation and apoptosis of breast cancer MDA-MB-231 cells through phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods:MDA-MB-231 cells were divided into control group(C group),cremastra appendiculata makino group(CAM group),insulin growth factor 1 group(IGF-1 group)and cremastra appendiculata makino+insulin growth factor 1 group(CAM+IGF-1 group).The cells of C group were not treated with drugs.The cells of CAM group were added with 10 mg/ml cremastra appendiculata makino.The cells of IGF-1 group were treated with 100μg/L IGF-1.The cells of CAM+IGF-1 group were treated with 10 mg/ml cremastra appendiculata makino and 100μg/L IGF-1.Thiazole blue(MTT)was used to measure the cell proliferation.The cell apoptosis was determined by flow cytometry and Hoechst 33258 staining.The cell-activated cysteine-containing aspartate hydrolase 3(Cleaved-Caspase-3),Bcl-2 related X protein(Bax),B-cell lymphoma factor(Bcl-2),PI3K,Phosphorylation of PI3K(p-PI3K),Akt and p-Akt protein levels were determined by Western blot.Results:Cremastra appendiculata makino inhibited the proliferation of MDA-MB-231 cells with dose and time dependent manner.Compared with C group,the OD value of MDA-MB-231 cells was decreased(P<0.05),the apoptosis rate was increased(P<0.05),the levels of Cleaved-Caspase-3 and Bax protein were increased(<0.05),the Bcl-2,p-PI3K and p-Akt protein levels were decreased(P<0.05)in CAM group.Compared with CAM group,the OD value of MDA-MB-231 cells was increased(P<0.05),the apoptosis rate was decreased(P<0.05),the levels of Cleaved-Caspase-3 and Bax protein were decreased(P<0.05),the Bcl-2,p-PI3K and p-Akt protein levels were elevated(P<0.05)in CAM+IGF-1 group.Conclusion:Cremastra appendiculata makino can inhibit breast cancer cell proliferation and induce breast cancer cell apoptosis,and its mechanism may be related to cremastra appendiculata makino inhibit the PI3K/Akt signaling pathway activation.
作者
兴伟
刘远
徐曌
徐志峰
宋易华(指导)
XING Wei;LIU Yuan;XU Zhao;XU Zhi-Feng;SONG Yi-Hua(Department 1 of Surgery,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第6期693-698,706,共7页
Chinese Journal of Immunology
基金
河北省中医药管理局科研计划项目(No.2018018)。
