核桃青皮提取物对乳腺癌MCF-7细胞增殖、凋亡及EGFR/MAPK信号通路的影响  

Effects of Walnut Green Husk Extract on Proliferation,Apoptosis and EGFR/MAPK Signal Pathway of Breast Cancer MCF-7 Cells

作　　者：张硕稳 李丹[1] 贺静[1] 杜志兴[1] 庞建民[1] 李瑞池[1] 刘永建[1] 郑丽华[2] ZHANG Shuowen;LI Dan;HE Jing;DU Zhixing;PANG Jianmin;LI Ruichi;LIU Yongjian;ZHENG Lihua(Health Management Center,the First Hospital of Hebei Medical University,Shijiazhuang 050000,China;Dept.of General Surgery,the First Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第一医院健康管理中心,石家庄050000 [2]河北医科大学第一医院普外科,石家庄050000

出　　处：《中国医院用药评价与分析》2022年第6期684-689,695,共7页Evaluation and Analysis of Drug-use in Hospitals of China

基　　金：河北省医学科学研究课题计划项目(No.20190484)。

摘　　要：目的:探讨核桃青皮提取物对乳腺癌MCF-7细胞增殖、凋亡及表皮生长因子受体(EGFR)/丝裂原活化蛋白激酶(MAPK)信号通路的影响。方法:采用细胞计数试剂盒-8(CCK-8)筛选干预乳腺癌MCF-7细胞的核桃青皮提取物合适浓度。将乳腺癌MCF-7细胞分为空白对照组、阳性对照组、核桃青皮提取物低浓度组、核桃青皮提取物高浓度组和核桃青皮提取物高浓度+EGFR激动剂(NSC228155)组。采用CCK-8和集落形成实验检测各组细胞增殖情况;采用TUNEL法和流式细胞仪检测各组细胞凋亡情况;采用蛋白质印迹法检测各组细胞中增殖、凋亡及EGFR/MAPK信号通路相关蛋白表达水平。结果:当核桃青皮提取物浓度≥20μg/mL时,细胞增殖率显著降低,故选择20、40μg/mL分别为核桃青皮提取物低浓度组、高浓度组的核桃青皮提取物处理浓度。与空白对照组相比,阳性对照组、核桃青皮提取物低浓度组和高浓度组细胞增殖率、细胞集落形成数量,细胞周期蛋白D1(CyclinD1)、增殖细胞核抗原(PCNA)和B细胞淋巴瘤2(Bcl-2)蛋白表达水平,磷酸化EGFR(p-EGFR)/EGFR、磷酸化胞外信号调节激酶(p-ERK)/胞外信号调节激酶(ERK)、磷酸化p38 MAPK(p-p38 MAPK)/p38 MAPK和磷酸化c-Jun氨基端蛋白激酶(p-JNK)/c-Jun氨基端蛋白激酶(JNK)蛋白比值均显著降低;TUNEL阳性细胞率、细胞凋亡率和Bcl-2相关X蛋白(Bax)表达水平均显著升高,差异均有统计学意义(P<0.05)。与核桃青皮提取物低浓度组相比,核桃青皮提取物高浓度组细胞增殖率、细胞集落形成数量,CyclinD1、PCNA和Bcl-2蛋白表达水平,p-EGFR/EGFR、p-ERK/ERK、p-p38 MAPK/p38 MAPK和p-JNK/JNK蛋白比值均降低;TUNEL阳性细胞率、细胞凋亡率和Bax蛋白表达水平均升高,差异均有统计学意义(P<0.05)。核桃青皮提取物高浓度组与阳性对照组上述指标的差异均无统计学意义(P>0.05)。与核桃青皮提取物高浓度组相比,核桃青皮提取物高�OBJECTIVE:To probe into the effects of walnut green husk extract on proliferation,apoptosis and epidermal growth factor receptor(EGFR)/mitogen activated protein kinase(MAPK)signaling pathway of breast cancer MCF-7 cells.METHODS:Cell counting kit-8(CCK-8)was used to screen the proper concentration of walnut husk extract interfering with breast cancer MCF-7 cells.The breast cancer MCF-7 cells were divided into the blank control group,positive control group,walnut husk husk extract low concentration group,walnut green husk extract high concentration group,walnut green husk extract high concentration+EGFR agonist(NSC228155)group.CCK-8 and colony formation experiment were used to detect cell proliferation in each group.TUNEL method and flow cytometry were used to detect cell apoptosis in each group.Western blot was used to detect the expression levels of proliferation,apoptosis and EGFR/MAPK signaling pathway related proteins of cells in each group.RESULTS:When the concentration of walnut green husk extract was≥20μg/mL,the cell proliferation rate decreased significantly,20μg/mL and 40μg/mL were selected as the walnut green husk extract low and high concentration treatment,respectively.Compared with the blank control group,the cell proliferation rate,the number of cell colonies,the expression levels of CyclinD1,proliferating cell nuclear antigen(PCNA)and B cell lymphoma-2(Bcl-2)protein,the ratio of phosphorylated-EGFR(p-EGFR)/EGFR,phosphorylated extracellular signal-regulated kinase(p-ERK)/extracellular signal-regulated kinase(ERK),phosphorylated-p38 MAPK(p-p38 MAPK)/p38 MAPK and phosphorylated c-Jun amino-terminal protein kinase(p-JNK)/c-Jun amino-terminal protein kinase(JNK)protein in the positive control group,the walnut green husk extract low concentration group and high concentration group decreased significantly,and the rate of TUNEL positive cells,the apoptosis rate,the expression level of Bcl-2 related X protein(Bax)increased significantly,with statistically significant differences(P<0.05).Compared with th

关 键 词：核桃青皮提取物 乳腺癌MCF-7细胞 增殖 凋亡 表皮生长因子受体/丝裂原活化蛋白激酶信号通路 

分 类 号：R96[医药卫生—药理学]