Interferon-α enhances sensitivity of human osteosarcoma U2OS cells to doxorubicin by p53-dependent apoptosis  

作　　者：Xiang-wei YUAN Xiao-feng ZHU Xiu-fang HUANG Pu-yi SHENG Ai-shan HE Zi-bo YANG Rong DENG Gong-kanFENG Wei-ming LIAO 

机构地区：[1]Department of Orthopedic Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China [2]State Key Laboratoryof Oncology in South China, Cancer Center, Sun Yat-sen University, Guangzhou 510060 [3]Department of Pathology, The AffiliatedJiangmen Hospital, Sun Yat-sen University, Jiangmen 529030, China

出　　处：《Acta Pharmacologica Sinica》2007年第11期1835-1841,共7页中国药理学报（英文版）

基　　金：grants from the National Natural Science Foundation of China(№ 30070766)

摘　　要：Aim： To determine whether interferon-or （IFNα） can enhance doxorubicin sensitivity in osteosarcoma cells and its molecular mechanism. Methods： Cell viability was evaluated using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. Apoptosis was studied using Flow cytometry analysis, Hoechst33258 staining, DNA fragmentation assay, as well as the activation of caspase-3 and poly （ADP-ribose） polymerase. Protein expression was detected by Western blotting. The dependence of p53 was determined using p53-siRNA transfection. Results： IFNα increased doxorubicin-induced cytotoxicity to a much greater degree through apoptosis in human osteosarcoma p53-wild U2OS cells, but not p53-mutant MG63 cells. IFNα markedly upregulated p53, Bax, Mdm2, and p21, downregulated Bcl-2, and activated caspase-3 and PARP cleavage in response to doxorubicin in U2OS cells. Moreover, the siRNA-mediated silencing of p53 significantly reduced the IFNα/doxorubicin combination-induced cytotoxicity and PARP cleavage. Conclusion： IFNα enhances the sensitivity of human osteosarcoma U2OS cells to doxorubicin by p53-dependent apoptosis. The proper combination with IFNα and conventional chemotherapeutic agents may be a rational strategy for improving the treatment of osteosarcoma with functional p53.Aim： To determine whether interferon-or （IFNα） can enhance doxorubicin sensitivity in osteosarcoma cells and its molecular mechanism. Methods： Cell viability was evaluated using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. Apoptosis was studied using Flow cytometry analysis, Hoechst33258 staining, DNA fragmentation assay, as well as the activation of caspase-3 and poly （ADP-ribose） polymerase. Protein expression was detected by Western blotting. The dependence of p53 was determined using p53-siRNA transfection. Results： IFNα increased doxorubicin-induced cytotoxicity to a much greater degree through apoptosis in human osteosarcoma p53-wild U2OS cells, but not p53-mutant MG63 cells. IFNα markedly upregulated p53, Bax, Mdm2, and p21, downregulated Bcl-2, and activated caspase-3 and PARP cleavage in response to doxorubicin in U2OS cells. Moreover, the siRNA-mediated silencing of p53 significantly reduced the IFNα/doxorubicin combination-induced cytotoxicity and PARP cleavage. Conclusion： IFNα enhances the sensitivity of human osteosarcoma U2OS cells to doxorubicin by p53-dependent apoptosis. The proper combination with IFNα and conventional chemotherapeutic agents may be a rational strategy for improving the treatment of osteosarcoma with functional p53.

关 键 词：Interferon-a DOXORUBICIN APOPTOSIS P53 OSTEOSARCOMA 

分 类 号：R4[医药卫生—临床医学]