雷帕霉素联合阿霉素对人骨肉瘤MG-63细胞增殖的影响及其机制  

Effects of Rapamycin and Adriamycin on proliferation of osteosarcoma MG-63 cells

作　　者：崔明星[1] 詹新立[2] 刘冲[2] 邓黎[2] 熊春翔[2] 刘会江[2] 

机构地区：[1]新乡医学院第一附属医院,河南新乡453100 [2]广西医科大学第一附属医院

出　　处：《山东医药》2017年第16期8-11,共4页Shandong Medical Journal

基　　金：广西自然科学基金资助项目(2012GXNSFAA05314;2014GXNSFAA118173);广西高校科学技术研究项目(YB2014073)

摘　　要：目的探讨雷帕霉素联合阿霉素对人骨肉瘤MG-63细胞(以下称骨肉瘤细胞)增殖的影响及其作用机制。方法取对数生长期骨肉瘤细胞,分别加入不同浓度的阿霉素(0.125、0.25、0.5、1.0、2.0 mg/L)和雷帕霉素(20、50、100 nmol/L)单独及联合作用,采用MTT法检测作用48 h的细胞增殖抑制率,并计算两药相互作用指数(CDI),评价两药相互作用性质。结果以100 nmol/L雷帕霉素与0.5 mg/L阿霉素联合作用后的细胞增殖抑制率最高,此时CDI<0.7,两种药物表现为显著协同作用。将对数生长期的骨肉瘤细胞分为五组,阿霉素组、雷帕霉素组分别加入0.5 mg/L阿霉素、100 nmol/L雷帕霉素,联合用药组加入0.5 mg/L阿霉素和100 nmol/L雷帕霉素,DMSO溶媒组加入等量DMSO,空白对照组不予处理。采用MTT法检测各组细胞增殖能力,连续记录5天。采用Real-time PCR法检测各组处理48 h缺氧诱导因子1α(HIF-1α)、血管内皮生长因子(VEGF)、基质金属蛋白酶2(MMP-2)mRNA相对表达量。结果各组作用第1、2天细胞增殖能力比较差异均无统计学意义(P均>0.05)。作用第3~5天,联合用药组细胞增殖能力均明显低于空白对照组、DMSO溶媒组、阿霉素组及雷帕霉素组(P均<0.05)。联合用药组HIF-1α、VEGF、MMP-2 mRNA相对表达量均明显低于空白对照组、DMSO溶媒组、阿霉素组、雷帕霉素组(P均<0.05)。结论雷帕霉素联合阿霉素可以协同抑制骨肉瘤细胞增殖;降低HIF-1α、VEGF、MMP-2表达可能是其作用机制。Objective To investigate the effects of Rapamycin （RAPA） and Adriamycin （ADM） on proliferation of osteosarcoma MG-63 cells and their mechanisms.Methods Human osteosarcoma MG-63 cells in logarithmic growth phase was treated with ADM solution of different concentrations （0.125, 0.25, 0.5, 1.0, 2.0 mg/L） and RAPA solution of different concentrations （20, 50, 100 nmol/L） alone or in combination.Then the cell inhibition rate was determined by MTT after 48 hours, and Coefficient of drug interaction （CDI） was calculated to evaluate the nature of the interaction between the two drugs.The cell inhibition rate was the highest when MG-63 cells were treated with 100 nmol/L RAPA combined with 0.5 mg/L ADM.At this moment, CDI was less than 0.7 and the two drugs showed a significant synergistic effect.According to the optimal combination concentration, we randomly divided MG-63 cells in logarithmic growth phase into five groups： ADM group （treated with 0.5 mg/L ADM）, RAPA group （treated with 100 nmol/L RAPA）, the combination group （treated with 0.5 mg/L ADM and 100 nmol/L RAPA）, DMSO solvent group （treated with the same concentration of DMSO） and the blank control group （without any treatment）.The cell proliferation ability was detected by MTT for 5 days.The expression levels of hypoxia-inducible factor-1α （HIF-1α）, vascular endothelial growth factor （VEGF） and matrix metalloproteinase 2 （MMP-2） mRNA were analyzed by real-time PCR at 48 h.Results There was no significant difference in cell proliferation ability among these groups on day 1 and 2 （all P〉0.05）.The cell proliferation ability of the combination group was lower than those of the blank control group, DMSO solvent group, ADM group and RAPA group on day 3-5 （all P〈0.05）.The relative expression of HIF-1α, VEGF and MMP-2 mRNA in the combination group was significantly lower than that in the blank control group, DMSO solvent group, ADM group and RAPA group （all P〈0.05）.Conclusion RAPA combined with

关 键 词：骨肉瘤 细胞增殖 雷帕霉素 阿霉素 药物敏感性 

分 类 号：R738.1[医药卫生—肿瘤]