摘要
目的探讨α干扰素(IFN-α)在人骨肉瘤U2OS和MG63细胞中对依托泊苷敏感性的作用及其机制。方法采用IFN-α和依托泊苷单独或联合处理骨肉瘤U2OS和MG63细胞72 h,应用流式细胞术和DNA Ladder检测细胞凋亡的变化,Western blot法检测PARP、p53、MDM2、Bax、Bcl-2的表达。结果流式细胞术表明IFN-α在p53正常的U2OS细胞中明显增强依托泊苷诱导的凋亡(P<0.05),却对依托泊苷诱导p53突变的MG63细胞凋亡无明显影响(P>0.05);DNA Ladder表明与单药组相比,IFN-α与依托泊苷联用72 h在U2OS细胞出现明显的DNA梯形条带,而在MG63细胞中无此现象;Western blot结果表明在U2OS细胞中联合用药组出现PARP的明显裂解活化,并且IFN-α明显增强依托泊苷引起的p53信号通路p53、MDM2、Bax等凋亡基因的表达,并抑制抗凋亡基因Bcl-2的表达,而在MG63细胞中则无此现象。结论 IFN-α能够通过激活p53依赖性信号通路增强依托泊苷诱导的骨肉瘤U2OS细胞凋亡,联合应用IFN-α和传统化疗药物如依托泊苷可能成为提高骨肉瘤化疗敏感性的有效途径。
Objective To study effect of IFN-α on the sensitivity to Etoposide in human osteosarcoma U2OS and MG63 cells with its molecular mechanisms. Methods U2OS and MG63 cells were treated with IFN-α and Etoposide, alone or in combination, for 72 h. Flow cytometry analysis and DNA Ladder were used to examine apoptosis. Western blot was used to determine expression of PARP, p53, Bax, Mdm2 and Bcl-2. Results IFN-α enhanced Etoposide-induced apoptosis in p53-wild U2OS cells（P 0.05）, but not p53-mutant MG63 cells（P 0.05）. IFN-α＋Etoposide combination result in obvious DNA Ladder in U2OS, but not MG63 cells and activation of PARP in U2OS cells. The enhanced apoptosis was associated with the accumulation of transcriptionally active p53 accompanied with the increased Bax and MDM2, as well as decreased Bcl-2, in U2OS cells, which was not observed in MG63 cells. Conclusion IFN-α enhances Etoposide-induced apoptosis in human osteosarcoma U2OS cells by activation of p53-dependent signal pathway. This work implies that rational combination with IFN-α and chemotherapy including Etoposide may be a useful strategy for enhancing the chemosensitivity of osteosarcoma.
出处
《中国医药导报》
CAS
2014年第7期4-7,14,共5页
China Medical Herald
基金
广东省医学科研基金立项课题(编号A2013756)
广东省医学科学技术研究课题(编号B2009249)
