徐长卿丹皮酚联合顺铂对骨肉瘤细胞迁移和凋亡及相关蛋白表达的影响  

Efficacy of Cynanchumpaniculatum Paeonol Combined with Cisplatin on Migration and Apoptosis of Osteosarcoma Cells and Expression of Related Proteins

作　　者：秦桂福[1,2] 吴琪 宋玉 吴高明 李俟琪 QIN Guifu;WU Qi;SONG Yu;WU Gaoming;LI Siqi(Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061,China;Hubei Institute of Traditional Chinese Medicine,Wuhan 430074,China;Clinical Skills Training Center of Hubei University of Traditional Chinese Medicine,Wuhan 430061,China;Haojiang Hospital,The First Affiliated Hospital of Shantou University Medical College,Shantou 515071,Guangdong China)

机构地区：[1]湖北省中医院,武汉430061 [2]湖北省中医药研究院 [3]湖北中医药大学临床技能实训中心 [4]汕头大学医学院第一附属医院濠江医院

出　　处：《中国中医骨伤科杂志》2021年第9期10-15,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：湖北省卫生健康委员会中医药科研项目(ZY2021Q012);湖北中医药大学“青苗计划”资助项目(2019ZZX021)。

摘　　要：目的:探讨徐长卿丹皮酚(Paeonol)联合顺铂对骨肉瘤MG-63细胞迁移和凋亡的影响及其作用机制。方法:培养骨肉瘤MG-63细胞,分别加入不同浓度的徐长卿丹皮酚(0.3,0.6,1.2mmol/L)和顺铂(4μmol/L)单独及联合作用,采用CKK-8法检测MG-63细胞增殖抑制率,并计算两药相互作用指数(CDI),评价两药相互作用性质,细胞划痕实验和Transwell实验检测MG-63细胞的迁移和侵袭能力,TUNEL法检测MG-63细胞凋亡情况,Western Blot法检测骨肉瘤MG-63细胞中Akt和mTOR蛋白表达及磷酸化水平。结果:徐长卿丹皮酚和顺铂联合给药可以显著抑制骨肉瘤细胞增殖,两药存在协同作用,CDI<1,其抑制作用与徐长卿丹皮酚浓度及作用时间相关,差异有统计学意义(P<0.05,P<0.01);徐长卿丹皮酚不同浓度联合顺铂给药可显著抑制骨肉瘤细胞迁移,并具有一定的浓度依赖性,差异有统计学意义(P<0.05,P<0.01);徐长卿丹皮酚联合顺铂可以显著抑制骨肉瘤细胞的侵袭能力,并具有一定的浓度依赖性;随着徐长卿丹皮酚浓度的增加,呈红色荧光凋亡细胞有明显增多的趋势,以徐长卿丹皮酚中、高浓度与顺铂联用最明显;徐长卿丹皮酚联合顺铂可显著抑制Akt、mTOR蛋白磷酸化水平,具有一定的浓度依赖性,差异有统计学意义(P<0.01)。结论:徐长卿丹皮酚增强顺铂抑制骨肉瘤MG-63细胞增殖、迁移、侵袭和诱导凋亡,可能通过抑制Akt、mTOR蛋白磷酸化,发挥协同抑瘤作用。Objective:To explore the efficacy of cynanchumpaniculatum paeonol combined with cisplatin on the migration and apoptosis of osteosarcoma MG-63cells and its mechanism.Methods:MG-63cells were cultured and treated with paeonol solution of different concentrations(0.3,0.6,1.2mmol/L)and cisplatin(4μmol/L)separately or in combination.The proliferation inhibition rate of MG-63cells was measured by CKK-8 method,and the two-drug interaction index(CDI)was calculated to evaluate the nature of the two-drug interaction.The migration and invasion ability of MG-63cells were detected by Scratch test and Transwell test.The apoptosis of MG-63 cells was detected by TUNEL.Western Blot was used to detect the protein expression and phosphorylation level of Akt and mTOR in osteosarcoma MG-63cells.Results:The results demonstrated that combination of paeonol and cisplatin could significantly inhibit the proliferation of osteosarcoma cells,and the two drugs had a synergistic effect due to CDI<1,which was related to the concentration and duration of paeonol(P<0.05,P<0.01).Different concentrations of paeonol combined with cisplatin could significantly inhibit the migration of osteosarcoma cells,and there was a certain concentration-dependent(P<0.05,P<0.01).The combination of paeonol and cisplatin could inhibit the invasion of osteosarcoma cells in a concentration-dependent manner,and with the increase of the concentration of paeonol,the combination of paeonol and cisplatin could significantly inhibit the protein phosphorylation of Akt and mTOR,and the combination of two drugs could significantly inhibit the protein phosphorylation of Akt and mTOR,and there was a certain concentration dependence(P<0.01).Conclusion:Paeonol can enhance the efficacies of cisplatin on the proliferation,migration,invasion and apoptosis of osteosarcoma MG-63cells,which may play a synergistic role by inhibiting the proteins phosphorylation of Akt and mTOR.

关 键 词：徐长卿丹皮酚 顺铂 骨肉瘤 迁移 凋亡 蛋白磷酸化水平 

分 类 号：R-33[医药卫生]