鹰嘴豆芽素A诱导人成骨肉瘤MG63细胞凋亡作用研究  

Biochanin-A induced apoptosis of human osteosarcoma MG-63 cells by regulating MAPK/apoptosis pathway

作　　者：黄乾 廖世杰[1,2] 林成森 刘云[1] 李波香 陈县祥 刘建宏 丁晓飞[1,2] HUANG Qi-an;LIAO Shi-jie;LIN Cheng-sen;LIU Yun;LI Bo-xiang;CHEN Xian-xiang;LIU Jian-hong;DING Xiao-fei(Department of Trauma Orthopedic and Hand Surgery,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Research Centre for Regenerative Medicine,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院骨科,广西南宁530021 [2]广西医科大学第一附属医院广西再生医学研究中心,广西南宁530021

出　　处：《中国矫形外科杂志》2021年第10期915-919,共5页Orthopedic Journal of China

基　　金：国家自然科学基金地区项目(编号:81960768);广西自然科学基金面上项目(编号:2019JJA140408,2017GXNSFAA198305);广西自然科学基金青年基金项目(编号:2018GXNSFBA281090)。

摘　　要：[目的]探讨植物雌激素(phytoestrogen)鹰嘴豆芽素A(biochanin-A,BA)诱导人成骨肉瘤MG-63细胞凋亡及其调控丝裂原活化蛋白激酶(mitogen-altivated protein kinase,MAPK)/凋亡信号通路的作用机制。[方法]用不同浓度的鹰嘴豆芽素A处理MG-63细胞,以MTT法检测细胞活性的抑制情况;以DAPI染色法和Annexin-V/PI流式细胞仪检测细胞的凋亡情况;应用Western blot法检测MG-63细胞中MAPK信号通路相关蛋白磷酸化和主要凋亡通路Bcl-2家族和caspase家族相关蛋白的表达情况。[结果]BA有效抑制MG-63细胞的细胞活性,并呈时间和剂量依赖效应,且可以诱导细胞凋亡。40μmol/L和80μmol/L的BA作用于MG-63细胞48 h,细胞凋亡率分别为(45.28±3.79)%和(82.51±5.23)%,高于正常组(5.31±3.14)%(P<0.05)。Western blot结果显示,BA可促进caspase-9、caspase-3和PARP发生剪切(P<0.05),并促进p-p38、p-JNK、Bax的表达(P<0.05),抑制Bcl-2的表达(P<0.05),并呈剂量依赖效应。[结论]鹰嘴豆芽素A可有效抑制人成骨肉瘤MG-63细胞的细胞活性,促进细胞凋亡,作用机制可能是通过激活MAPK/凋亡信号通路来实现。[Objective] To explore the mechanism of phytoestrogen biochanin-A(BA) inducing apoptosis of human osteosarcoma MG-63 cells by mitogen-activated protein kinase(MAPK)/apoptotic signaling pathway. [Methods] MG-63 cells were treated with different concentrations of BA. MTT assay was used to detect the inhibition of cell viability. DAPI staining and Annexin V/PI flow cytometry were conducted to measure cell apoptosis. In addition, western blot was used to detect the phosphorylation of MAPK signaling pathway related proteins and the expression of Bcl-2 and caspase family related proteins in MG-63 cells. [Results] The BA effectively inhibited the cell viability of MG-63 cells in a time-and dose-dependent manners, and could induce MG-63 cells apoptosis. Approximately 40 μmol/L and 80μmol/L BA was added on MG-63 cells for 48 hours, and the cell apoptosis rates were(45.28±3.79)% and(82.51±5.23)% respectively,which both were higher than that of the control group(5.31±3.14)%(P<0.05). As results of western blot, the BA promoted the cleavage of caspase-9, caspase-3 and PARP(P<0.05), promoted the expression of p-p38, p-JNK and Bax(P<0.05), whereas inhibited the expression of Bcl-2(P<0.05) in a dose-dependent manner. [Conclusion] The biochanin-A does effectively inhibit the cell viability of human osteosarcoma MG-63 cells and promote cell apoptosis, and the mechanism of action may be achieved by activating MAPK/apoptotic signaling pathway.

关 键 词：鹰嘴豆芽素A 骨肉瘤细胞MG-63 丝裂原活化蛋白激酶(MAPK) 凋亡 

分 类 号：R318[医药卫生—生物医学工程]