淫羊藿素联合阿霉素对人骨肉瘤MG-63细胞抑制作用的体外研究  

Inhibition of Combination of Icaritin and Doxorubicin on Human Osteosarcoma MG-63 Cells in vitro

作　　者：林思文[1,2] 李雪芹[3] 刘素云[1] 施剑明 徐浚怀 毛龙火 殷明[1] 

机构地区：[1]南昌大学第二附属医院骨一科,南昌330006 [2]南昌大学研究生院医学部,南昌330006 [3]九江学院基础医学院,江西332000

出　　处：《中国中西医结合杂志》2016年第6期729-734,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81160226)

摘　　要：目的探讨体外淫羊藿素联合阿霉素对人骨肉瘤MG-63细胞的抑制作用及分子机制。方法设置对照组及不同浓度梯度的淫羊藿素组(10、20、40、80、160μmol/L)、阿霉素组(1、2、4、8、16μg/m L)、联合组(淫羊藿素+阿霉素:20μmol/L+1μg/m L、20μmol/L+2μg/m L、20μmol/L+4μg/m L、40 mol/L+1μg/m L、40μmol/L+2μg/m L、40μmol/L+4μg/m L、80μmol/L+1μg/m L、80μmol/L+2μg/m L、80μmol/L+4μg/m L),分别培养骨肉瘤MG-63细胞用倒置相差显微镜观察各组药物作用24、48 h后细胞形态;CCK-8法测定各组的细胞增殖抑制率并计算半数抑制浓度;AnnexinⅤ-FITC/PI双染料流式细胞分析技术测定各组的细胞凋亡率;RT-PCR检测bcl-2、caspase-3及p21表达。结果淫羊藿素及阿霉素可明显抑制MG-63细胞增殖,随着淫羊藿素的浓度从10μmol/L逐渐增加至160μmol/L,细胞增殖抑制率也随之从(9.67±3.62)%逐渐增加至(89.18±9.66)%,二者的半数抑制浓度分别为:46.93μmol/L、3.87μg/m L;淫羊藿素与阿霉素可致细胞早期及晚期凋亡,使bcl-2基因表达下调、caspase-3、p21基因表达上调(P<0.05);以上改变联合组较淫羊藿素组及阿霉素组更明显(P<0.05)。结论淫羊藿素联合阿霉素对抑制人骨肉瘤MG-63细胞增殖具有协同或相加效应,其作用机制可能与调控bcl-2、caspase-3及p21基因有关。Objective To explore the inhibition and molecular mechanism of icaritin（ICT） combined doxorubicin（DOX） on human osteosarcoma M G- 63 cells in vitro.Methods The control group,ICT groups（10,20,40,80,and 1 60 μmol/L）,DOX groups（1,2,4,8,and 1 6μg/mL）,and combination groups（20 μmol/L ICT ＋ 1 μg/mL DOX,20 μmol/L ICT ＋ 2 μg/mL DOX,20 μmol/L ICT ＋ 4 μg/mL DOX,40 μmol/L ICT ＋ 1 μg/mL DOX,40 μmol/L ICT ＋ 2μg/mL DOX,40 μmol/L ICT ＋ 4 μg/mL DOX,80 μmol/L ICT ＋ 1 μg/mL DOX,80 μmol/L ICT ＋2 μg/mL DOX,80 μmol/L ICT ＋4 μg/mL DOX） were set up.Human osteosarcoma MG-63 cells were respectively cultured and their effects on morphological changes were observed using inverted phase contrast microscope after 24- and 48-h intervention.The cell proliferation inhibition rate of each group was determined using CCK-8,and IC50 calculated.The MG-63 apoptosis rate was detected using Annexin V-FITC/PI double dye flow cytometry.Expression levels of bcl-2,caspase-3,and p21 were detected using RT-PCR.Results ICT and DOX could obviously inhibit the proliferation of MG-63 cell.Along with ICT concentration increasing from 10 μmol/L to 160 μmol/L,the cell proliferation inhibition rate also increased gradually from 9.67%±3.62%to 89.18%±9.66%.The IC50 was 46.93 μmol/L and 3.87 μg/mL respectively.ICT and DOX could cause either early or late stage apoptosis,down-regulate Bcl-2 gene expression,and up-regulate gene expressions of Caspase-3 and p21 respectively（P 〈0.05）.Aforesaid changes were more obviously seen in combination groups than in ICT groups and DOX groups（P 〈0.05）.Conclusion ICT combined DOX had additive or synergistic inhibition effect for the proliferation of osteosarcoma MG-63 cells,which might be related with regulating gene expressions of bcl-2,caspase-3,and p21.

关 键 词：淫羊藿素 阿霉素 骨肉瘤MG-63细胞 

分 类 号：R285[医药卫生—中药学]