姜黄素对体内外骨肉瘤细胞增殖和侵袭的影响及其机制研究  

Effects of Curcumin on in vitro and in vivo Proliferation and Invasion of Osteosarcoma Cells and Its Mechanism Study

作　　者：颜泉[1] YAN Quan(Dept.of Orthopaedics,Luzhou Municipal People’s Hospital,Sichuan Luzhou 646000,China)

机构地区：[1]泸州市人民医院骨科,四川泸州646000

出　　处：《中国药房》2018年第7期918-922,共5页China Pharmacy

基　　金：四川省科技厅资助项目(No.2015jy0224)

摘　　要：目的:研究姜黄素对骨肉瘤细胞增殖和侵袭的影响及其机制。方法:体外细胞试验分为对照组(二甲基亚砜),姜黄素低、中、高浓度组(10、20、40μmol/L),姜黄素高浓度+内源性微小核糖核酸-21模拟物(agomiR-21)组和姜黄素高浓度+agomiR-21阴性对照(NC)组,分别采用CCK-8和Transwell小室法检测各组人骨肉瘤细胞U-2OS和MG-63的增殖[光密度(OD值)]和侵袭能力(穿膜细胞数),分别采用实时荧光定量聚合酶链式反应法和Western blot法检测各组细胞中miR-21及其靶基因PDCD4蛋白的表达。体内实验将MG-63细胞接种至小鼠体内,然后将小鼠分为空白对照组(生理盐水)、姜黄素组(20 mg/kg)、姜黄素+agomiR-21NC组(20 mg/kg姜黄素溶液+50 mg/kg agomiR-21 NC溶液)、姜黄素+agomiR-21组(20 mg/kg姜黄素溶液+50 mg/kg agomiR-21溶液),每组8只,分别于瘤内注射相应药物,每天1次,连续给药7 d。测量给药后20 d内的肿瘤体积变化,检测第20天瘤组织中miR-21和PDCD4蛋白的表达。结果:与对照组比较,姜黄素低、中、高浓度组U-2OS、MG-63细胞的OD值和穿膜细胞数均明显减小(P<0.05或P<0.01);其中姜黄素中、高浓度组U-2OS、MG-63细胞中miR-21的表达明显减弱(P<0.05或P<0.01),PDCD4蛋白表达明显增强(P<0.01)。与姜黄素高浓度+agomiR-21 NC组比较,姜黄素高浓度+agomiR-21组U-2OS、MG-63细胞的OD值和穿膜细胞数均明显增加(P<0.01)。与空白对照组比较,姜黄素组和姜黄素+agomiR-21 NC组小鼠给药5 d后肿瘤体积明显减小(P<0.05或P<0.01);瘤组织中miR-21表达明显减弱(P<0.01),PDCD4蛋白表达明显增强(P<0.01)。与姜黄素+agomiR-21 NC组比较,姜黄素+agomiR-21组小鼠给药5 d后肿瘤体积明显增加(P<0.05或P<0.01);瘤组织中miR-21表达明显增强(P<0.01),PDCD4蛋白表达明显减弱(P<0.01)。结论:姜黄素可抑制人骨肉瘤细胞U-2OS、MG-63的增殖和侵袭,其机制可能与下调miR-21表达有关。OBJECTIVE:To investigate the effects of curcumin on proliferation and invasion of osteosarcoma cells and its mechanism.METHODS:In in vitro cell assay,osteosarcoma cells were divided into control group(dimethyl sulfoxide),curcumin low-concentration,medium-concentration and high-concentration groups(10,20,40μmol/L)and curcumin high-concentration+endogenous miR-21 simulacrum(agomiR-21)group,curcumin high-concentration+agomiR-21 negative control(NC)group.The proliferation(OD value)and invasion ability(transmembrane cells number)of human osteosarcoma cells U-2OS and MG-63 were detected by CCK-8 and Transwell chamber method;the expression of miR-21 and its target gene PDCD4 protein were detected by real-time fluorescence quantitative PCR and Western blot assay.In in vivo cell assay,MG-63 cells were inoculated in mice,and the mice were divided into blank control group(normal saline),curcumin group(20 mg/kg),curcumin+agomiR-21 NC group(20 mg/kg curcumin solution+50 mg/kg agomiR-21 NC solution),curcumin+agomiR-21 group(20 mg/kg curcumin solution+50 mg/kg agomiR-21 solution)with 8 mice in each group;they were given intratumoral injection of relevant medicine once a day,for consecutive 7 d.The volume changes of the tumor were measured within 20 d after medication,and the expression of miR-21 and PDCD4 protein in tumor tissue were detected on 20th day.RESULTS:Compared with control group,the OD value and transmembrane cells number of U-2OS and MG-63 cells were decreased significantly in curcumin low-concentration,medium-concentration and high-concentration groups(P<0.05 or P<0.01);the expression of miR-21 was weakened significantly in curcumin medium-concentration and high-concentration groups of U-2OS and MG-63 cells(P<0.05 or P<0.01),while the expression of PDCD4 protein was strengthened significantly(P<0.01).Compared with curcumin high-concentration+agomiR-21 NC group,the OD value and transmembrane cells number of U-2OS and MG-63 cells were strengthened significantly in curcumin high-concentration+agomiR-21 group(P<0.01).

关 键 词：姜黄素 微小核糖核酸-21 骨肉瘤细胞 增殖 侵袭 体内 体外 

分 类 号：R738.1[医药卫生—肿瘤]