α干扰素增强骨肉瘤细胞对依托泊苷敏感性实验研究.docVIP

α干扰素增强骨肉瘤细胞对依托泊苷敏感性实验研究 　　α干扰素增强骨肉瘤细胞对依托泊苷敏感性的 　　实验研究 　　黄秀芳1 原向伟2 　　1.广东省江门市中心医院病理科，广东江门 529030；2.广东省江门市中心医院骨科，广东江门 529030 　　[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U2O啊、胞中明显增强依托泊苷诱导的凋亡（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。 　　[关键词] α干扰素；依托泊苷；骨肉瘤；凋亡；p53 　　[中图分类号] R738.1 [文献标识码] A [文章编号] 1673-7210（2014）03（a）-0004-05 　　Experimental study of IFN-α for enhancing sensitivity to Etoposide in human osteosarcoma cells 　　HUANG Xiufang1 YUAN Xiangwei2 　　1.Department of Pathology， Jiangmen Central Hospital， Guangdong Province， Jiangmen 529030， China； 2.Department of Orthopaedic Surgery， Jiangmen Central Hospital， Guangdong Province， Jiangmen 529030， China 　　[Abstract] 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）. 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 p