骨肉瘤常用化疗药物作用的分子机制及用药方法选择  

The experimental study on the molecular mechanisms of drugs commonly used in treatment of osteosarcoma as well as the choice of medication

作　　者：徐袁秋[1] 刘思源[2] 张卫华[2] 刘星[2] 林亮[2] 许新明[1] 

机构地区：[1]河北医科大学第三医院放疗科,石家庄市050051 [2]河北医科大学第三医院骨肿瘤科,石家庄市050051

出　　处：《河北医药》2015年第14期2100-2103,共4页Hebei Medical Journal

摘　　要：目的研究常用骨肉瘤化疗药物作用的分子机制,对临床用药方式提供指导。方法将骨肉瘤细胞混悬液与不同浓度吡柔比星、顺铂、卡铂、甲氨蝶呤、异环磷酰胺分别作用24、48、72 h后,用倒置显微镜观察细胞生长情况。用MTT法测定每一孔的吸光度值OD,制作生长曲线,并求出药物的IC50。用IC50浓度的药物分别作用于骨肉瘤细胞24、48和72 h后用流式细胞仪进行细胞周期分析,了解细胞的周期分布及细胞的增殖活性。免疫印迹技术观察细胞的PCNA、Bcl-2、Bax、cyclin D1、cyclin E的表达。结果在倒置显微镜下观察不同药物浓度作用于的骨肉瘤细胞,比较细胞死亡数量于时间关系。MTT实验:不同时间组及不同浓度组与对照组相比,对MG-63细胞的生长抑制作用差异均具有显著性(P<0.05),呈现不同剂量-时间效应关系。细胞周期分析:在细胞各期可见明显凋亡峰,细胞周期各期所占比例无明显变化,抑制作用呈剂量依赖性。蛋白免疫印迹:PCNA在不同药物组骨肉瘤表达与对照组相比明显不同,不同药物组Bax表达与对照组明显增加,Bcl-2表达减少。cyclin D1、cyclin E的表达明显减少。结论吡柔比星对MG-63细胞的抑制率和时间关系密切,和浓度有一定的关系。同样剂量吡柔比星5 d的效果比3 d好。卡铂具有与顺铂类似的效应,但卡铂如果替代顺铂在化疗中的作用,卡铂的量必须是顺铂的25倍以上。甲氨蝶呤作用于骨肉瘤细胞MG-63后,随着药物浓度的增加和时间的延长,作用逐渐增强,尤其是用量和效果有明显关系。异环磷酰胺在体外对骨肉瘤细胞无明显抑制作用。Objective To investigate the molecular mechanisms of drugs commonly used in treatment of osteosarcoma in order to provide guidance for clinical medication.Methods The osteosarcoma cells-MG-63 were treated with different concentrations of pirarubicin,cisplatin,carboplatin,methotrexate,ifosfamide for 24 h,48h,72 h,respectively,then the cell growth state was observed under microscope.The MTT assay was used to make growth curve,and the cell cycle was analyzed by flow cytometry.The expression levels of PCNA,Bcl-2,Bax,cyclin D1,cyclin E were detected by Western blotting.Results There were significant differences in the growth suppression effects on MG-63 between different concentration groups,different action time groups and control group（P〈0.05）,with a dose-dependent and time-dependent correlation.The obvious apoptotic peak could be observed in different cell cycles,however,there was no obvious difference in the proportion of apoptosis cells among different cell cycles,moreover,the suppression effects were dose-dependent.There were significant differences in the expression levels of PCNA between different drugs groups and control group.As compared with those in control group,the expression levels of Bax in different drugs groups were obviously increased,however,those of Bcl-2 were obviously decreased,so were those of cyclin D1 and cyclin E.Conclusion The inhibition rate of pirarubicin on MG-63 cells is closely correlated to action time and to drug concentration at some extent.The efficiency of carboplatin is the same as that of cisplatin,but the dose of carboplatin must be 25 times more than that of cisplatin.The inhibition rate of methotrexate on MG-63 cells is increased with the increase of time and dosage.Ifosfamide has no obvious inhibition effect on osteosarcoma cells in vitro.

关 键 词：吡柔比星 顺铂 卡铂 甲氨蝶呤 异环磷酰胺 骨肉瘤细胞株MG-63 流式细胞分析 蛋白免疫印迹 

分 类 号：R730.266[医药卫生—肿瘤]