丹皮酚对人骨肉瘤MG-63细胞增殖、迁移和CXCR4/STAT3通路的影响  

Effect of paeonol on proliferation,migration,and CXCR4/STAT3 pathway of human osteosarcoma MG-63 cells

作　　者：吴琪 陈建锋 李浩 文峰 WU Qi;CHEN Jianfeng;LI Hao;WEN Feng(Needle Bone Laboratory,Clinical Skills Training Center,Hubei University of Traditional Chinese Medicine,Wuhan 430000,China;Department of Orthopaedics,Affiliated Hospital,Hubei University of Traditional Chinese Medicine,Wuhan 430000,China)

机构地区：[1]湖北中医药大学临床技能实训中心针骨实验室,湖北武汉430000 [2]湖北中医药大学附属医院骨科,湖北武汉430000

出　　处：《吉林大学学报（医学版）》2023年第5期1202-1209,共8页Journal of Jilin University：Medicine Edition

基　　金：湖北省教育厅科研计划重点项目(D20212004)。

摘　　要：目的:探讨丹皮酚对人骨肉瘤(OS) MG-63细胞增殖、迁移和趋化因子受体4(CXCR4)/信号传导与转录激活因子3 (STAT3)通路的影响,并阐明其可能的作用机制。方法:体外培养MG-63细胞,分为对照组(不给予药物干预)和不同浓度丹皮酚组(给予10、20、40、80、160和320 mg·L^(-1)丹皮酚)。采用CCK-8法检测各组细胞存活率并选择半数抑制浓度(IC50)值作为后续实验药物浓度。将MG-63细胞分为对照组和丹皮酚(215.8 mg·L^(-1))组,采用Western blotting法检测各组细胞中CXCR4、白细胞介素6 (IL-6)、磷酸化STAT3 (p-STAT3)和STAT3蛋白表达水平。将MG-63细胞分为对照组(不给予药物干预)、丹皮酚组(给予215.8 mg·L^(-1)丹皮酚)、丹皮酚+空载组(给予215.8 mg·L^(-1)丹皮酚+空载质粒)和丹皮酚+过表达组(给予215.8 mg·L^(-1)丹皮酚+CXCR4过表达质粒)。采用细胞划痕实验检测各组细胞划痕愈合率,克隆形成实验检测各组细胞克隆形成率,Western blotting法检测各组细胞中CXCR4、IL-6、p-STAT3和STAT3蛋白表达水平。结果:与对照组比较,40、80、160和320 mg·L^(-1)丹皮酚组MG-63细胞存活率降低(P<0.05),丹皮酚IC50值为215.8mg·L^(-1)。与对照组比较,丹皮酚(215.8mg·L^(-1))组MG-63细胞中CXCR4、IL-6和p-STAT3蛋白表达水平降低(P<0.05)。与对照组比较,丹皮酚组和丹皮酚+空载组MG-63细胞划痕愈合率及细胞克隆形成率降低(P<0.05),细胞中CXCR4、IL-6和p-STAT3蛋白表达水平降低(P<0.05);与丹皮酚组比较,丹皮酚+空载组MG-63细胞划痕愈合率、细胞克隆形成率和细胞中CXCR4、IL-6、p-STAT3及STAT3蛋白表达水平差异无统计学意义(P>0.05);与丹皮酚组比较,丹皮酚+过表达组MG-63细胞划痕愈合率和细胞克隆形成率升高(P<0.05),细胞中CXCR4、IL-6和p-STAT3蛋白表达水平升高(P<0.05)。结论:丹皮酚能抑制MG-63细胞的增殖、迁移和克隆形成,其机制可能与调控CXCR4/STAT3信号通路有关。Objective:To discuss the effect of paeonol on the proliferation,migration and chemokinereceptor4(CXCR4)/signal transducer and activators of transcription3(STAT3)pathway of the human osteosarcoma(OS)MG-63 cells,and to clarify its possible mechanism.Methods:The MG-63 cells were cultured in vitro and divided into control group(without drug intervention)and different concentrations of paeonol groups(given 10,20,40,80,160,and 320 mg·L^(-1) paeonol,respectively).The survival rates of the MG-63 cells in various groups were detected by CCK-8 method and the half inhibitory concentration(IC50)value was selected as the drug concentration for subsequent experiment.The MG-63 cells were divided into control group(without drug treatment)and paeonol group(given 215.8 mg·L^(-1) paeonal).The expression levels of CXCR4,interleukin 6(IL-6),and phosphorylated STAT3(p-STAT3),and STAT3 proteins in the MG-63 cells in various groups were detected by Western blotting method.The MG-63 cells were divided into control group,paeonol group,paeonol+empty group(given 215.8 mg·L^(-1) paeonol+empty plasmid),and paeonol+overexpression group(given 215.8 mg·L^(-1) paeonol+CXCR4 over-expression plasmid).The scratch healing rates of the MG-63 cells in various groups were detected by cell scratch experiment,the clone formation rates the MG-63 cells in various groups were detected by cloning formation experiment,and the expression levels of CXCR4,IL-6,p-STAT3,and STAT3 proteins in the MG-63 cells in various groups were detected by Western blotting method.Results:Compared with control group,the survival rates of the MG-63 cells treated in 40,80,160,and 320 mg·L^(-1) paeonol groups were decreased(P<0.05),and the IC50 value of paeonol was 215.8 mg·L^(-1).Compared with control group,the expression levels of CXCR4,IL-6,and p-STAT3 proteins in the MG-63 cells in paeonol group were decreased(P<0.05).Compared with control group,the scratch healing rates and clone formation rates of the MG-63 cells in paeonol group and paeonol+empty group were decreased(P<0

关 键 词：丹皮酚 人骨肉瘤细胞 细胞增殖 趋化因子受体4 信号传导与转录激活因子3 

分 类 号：R273[医药卫生—中西医结合] R738.1[医药卫生—中医肿瘤科]