低剂量吡柔比星协同TRAIL高效诱导人骨肉瘤细胞MG-63凋亡的实验研究  

Apoptosis of Osteosarcoma Cell MG-63 High Effectively Induced by A Combination of TRAIL and Low Does of THP : An Experiment Study

作　　者：乔晓晨[1] 吕智[1] 冯毅[1] 齐环宇[1] 薛小云[1] 贾鹏[1] 

机构地区：[1]山西医科大学第二临床医学院骨科,山西太原030001

出　　处：《中国现代医生》2011年第6期8-10,共3页China Modern Doctor

摘　　要：目的探讨肿瘤坏死因子相关诱导凋亡配体(TRAIL)与化疗药物吡柔比星(THP)联合应用是否存在协同诱导人骨肉瘤细胞MG-63凋亡的作用,并初步分析其可能存在的分子机制。方法将不同浓度的TRAIL与THP单独或者联合应用于常规培养的MG-63细胞系,24h后采用MTT法测定细胞抑制率,于倒置相差显微镜下观察细胞的形态学改变,流式细胞术检测细胞凋亡比例,RT-PCR检测药物作用后相关基因的表达水平。结果 (1)TRAIL与THP均能抑制MG-63细胞增殖,但MG-63细胞对TRAIL不敏感I,C50>10μg/mL;对THP相对敏感I,C50<10μg/mL;(2)亚毒性浓度THP与不同浓度TRAIL联合,均呈现出高效协同作用(P<0.05),协同因子均大于1;(3)倒置相差显微镜及流式细胞术检测证实协同性杀伤作用主要通过诱导细胞凋亡实现;(4)RT-PCR显示两种药物联用后死亡受体DR4及DR5表达水平得到明显的增高。结论亚毒性浓度THP与TRAIL联用表现出高效诱导人骨肉瘤MG-63细胞凋亡的作用,死亡受体DR4及DR5在药物作用前后表达水平的改变可能参与了这一凋亡过程。Objective To investigate whether TRAIL combined with THP can enhance inducing human osteosarcoma cell MG-63 apoptosis or not,and to analyze the potential molecular mechanism initially. Methods Different concentrations of TRAIL was used alone or in combination with THP on conventional cultured cell line MG-63 ,cell inhibition rate was evaluated by MTT Method after 24 hours, the morphological changes of cells observed under the inverted microscope ,the proportion of apoptosis detected by flow cy- tometry assay,the related gene expression levels detected by RT-PCR. Results （1）TRAIL and THP can inhibit the proliferation of MG-63 cells, MG-63 cell is not sensitive to TRAIL, ICSO 〉 10μg/mL, but relatively sensitive to THP, IC50 〈 10 μ g/mL （2）Subtoxic concentration of THP combined with different concentrations of TRAIL showed a high synergy（P〈0.05）,all eofaetors were more than 1. （3）Observation under the inverted microscope and detection by flow cytometry assay confirmed synergistic killing effect achieved mainly through the induction of apoptosis.（4）RT-PCR showed death receptors DR4 and DR5 expression levels were significantly higher after drug. Conclusion TRAIL can effectively induce human osteosareoma cells MG-63 apoptosis in combination of subtoxie does of THP, the change of death receptors DR4 and DR5 expession levels before and after drug may be involved in apoptosis.

关 键 词：肿瘤坏死因子相关诱导凋亡配体 吡柔比星 细胞凋亡 骨肉瘤 

分 类 号：R738.1[医药卫生—肿瘤]