OBJECTIVE Low dose of actinomycin D(LDAct D)was reported as a potent P53 activator and protected normal proliferating cells during anti-mitotic chemotherapy.However,the mechanism of LDAct D on P53 activation is still ...OBJECTIVE Low dose of actinomycin D(LDAct D)was reported as a potent P53 activator and protected normal proliferating cells during anti-mitotic chemotherapy.However,the mechanism of LDAct D on P53 activation is still undetermined.In this study,the mechanism of LDAct D on the synergistic antitumor effect for cisplatin(CDDP)and P53 reactivation in KB cells was studied in detail.METHODS Cell viability was determined by MTT and LDH release.Apoptosis was determined by AnnexinⅤ-FITC/PI staining.Mitochondrial membrane potential(MMP)was detected by JC-1 stain-ing.Expression of P53,PARP,BAX,BCL-XL,PUMA,MDM2 and MDMX was detected by Western blotting(WB)and/or immunofluorescence(IF).P53-MDM2 complex was detected by ELISA.Molecular docking of receptor MDM2 and MDMX with actinomycin D(ACTD)was analyzed by Discovery Studio.RESULTS Compared with CDDP alone,P53 expression and the cytotoxicity on KB cells was significantly increased by the combination therapy.P53 regulatory proteins were increased while MMP was decreased.Meanwhile,knockdown of PUMA(P53 upregulated modulator of apoptosis)efficiently blocked the synergistic effect of LDAct D to CDDP.P53 activation was found to be accompanied with the increase of MDMX but not MDM2.Meanwhile,MDM2-P53 complex in KB cells was significantly decreased by LDAct D.Docking of both receptor MDM2 and MDMX with ACTD exhibited well established bonds with nearby amino acid residues.CONCLUSION LDAct D was probably an inhibitor of both MDM2 and MDMX.The synergistic effects of LDAct D for CDDP on KB cells depended on its effect on reactivating P53 and PUMA mediated mitochondrial apoptosis.展开更多
Aim To study the synergistic effect and mechanism of actinomycin D to the anti-cancer activity of cispl- atin on KB cells. Methods Cytotoxicity of actinomycin D on KB cells was evaluated by MTT and LDH Assay. Apoptosi...Aim To study the synergistic effect and mechanism of actinomycin D to the anti-cancer activity of cispl- atin on KB cells. Methods Cytotoxicity of actinomycin D on KB cells was evaluated by MTT and LDH Assay. Apoptosis was detected by Flow cytometry （FCM）. Expression of p53, PUMA, Bax, Bcl-2 and Bcl-xl was detected by WB （Western blot） or IF （immunofluorescence staining）. The translocation of PUMA, Bax, Bcl-2 and Bcl-xl was detected by confocal microscope. PUMA knockdown was achieved by PUMA siRNA. Results Actinomycin D synergistically enhanced the cytotoxicity of cisplatin on KB cells. Time-dependent increasing of PUMA and p53 in- duced by actinomycin D was accompanied by the translocation of PUMA and Bax/Bcl-xl in KB cells. Knockdown of PUMA effectively blocked the synergistic effect of actinomycin D to cisplatin. Conclusion Actinomycin D effi- ciently enhanced the anti-cancer activity of cisplatin on KB cells. Up-regulation of PUMA by actinomycin D is like- ly responsible for the observed synergistic effect between the two drugs. Combination of actinomycin D and cisplatin may lead to an effective cancer treatment strategy.展开更多
AMD推出基于RV670核心的系列产品获得市场好评。其系列中的顶级型号Radeon HD 3870的3D性能较上一代旗舰R600核心的Radeon HD 2900 XT有较大提升。不过从我们以往的测试来看，Radeon HD 3870的3D性能并不如ONVIDIA的顶级产品GeForce 88...AMD推出基于RV670核心的系列产品获得市场好评。其系列中的顶级型号Radeon HD 3870的3D性能较上一代旗舰R600核心的Radeon HD 2900 XT有较大提升。不过从我们以往的测试来看，Radeon HD 3870的3D性能并不如ONVIDIA的顶级产品GeForce 8800 Ultra。基于R680核心的新旗舰产品——Radeon HD 3870×2，就是在这样背景下诞生，AMD意在靠它重夺显卡旗舰霸主。展开更多
基金The project supported by Ministry of Science and Technology Project of International Cooperation(2011DFR31240)National Science and Technology Major Projects″Major Drug Discovery(″2012ZX09301002001001)
文摘OBJECTIVE Low dose of actinomycin D(LDAct D)was reported as a potent P53 activator and protected normal proliferating cells during anti-mitotic chemotherapy.However,the mechanism of LDAct D on P53 activation is still undetermined.In this study,the mechanism of LDAct D on the synergistic antitumor effect for cisplatin(CDDP)and P53 reactivation in KB cells was studied in detail.METHODS Cell viability was determined by MTT and LDH release.Apoptosis was determined by AnnexinⅤ-FITC/PI staining.Mitochondrial membrane potential(MMP)was detected by JC-1 stain-ing.Expression of P53,PARP,BAX,BCL-XL,PUMA,MDM2 and MDMX was detected by Western blotting(WB)and/or immunofluorescence(IF).P53-MDM2 complex was detected by ELISA.Molecular docking of receptor MDM2 and MDMX with actinomycin D(ACTD)was analyzed by Discovery Studio.RESULTS Compared with CDDP alone,P53 expression and the cytotoxicity on KB cells was significantly increased by the combination therapy.P53 regulatory proteins were increased while MMP was decreased.Meanwhile,knockdown of PUMA(P53 upregulated modulator of apoptosis)efficiently blocked the synergistic effect of LDAct D to CDDP.P53 activation was found to be accompanied with the increase of MDMX but not MDM2.Meanwhile,MDM2-P53 complex in KB cells was significantly decreased by LDAct D.Docking of both receptor MDM2 and MDMX with ACTD exhibited well established bonds with nearby amino acid residues.CONCLUSION LDAct D was probably an inhibitor of both MDM2 and MDMX.The synergistic effects of LDAct D for CDDP on KB cells depended on its effect on reactivating P53 and PUMA mediated mitochondrial apoptosis.
文摘Aim To study the synergistic effect and mechanism of actinomycin D to the anti-cancer activity of cispl- atin on KB cells. Methods Cytotoxicity of actinomycin D on KB cells was evaluated by MTT and LDH Assay. Apoptosis was detected by Flow cytometry （FCM）. Expression of p53, PUMA, Bax, Bcl-2 and Bcl-xl was detected by WB （Western blot） or IF （immunofluorescence staining）. The translocation of PUMA, Bax, Bcl-2 and Bcl-xl was detected by confocal microscope. PUMA knockdown was achieved by PUMA siRNA. Results Actinomycin D synergistically enhanced the cytotoxicity of cisplatin on KB cells. Time-dependent increasing of PUMA and p53 in- duced by actinomycin D was accompanied by the translocation of PUMA and Bax/Bcl-xl in KB cells. Knockdown of PUMA effectively blocked the synergistic effect of actinomycin D to cisplatin. Conclusion Actinomycin D effi- ciently enhanced the anti-cancer activity of cisplatin on KB cells. Up-regulation of PUMA by actinomycin D is like- ly responsible for the observed synergistic effect between the two drugs. Combination of actinomycin D and cisplatin may lead to an effective cancer treatment strategy.
文摘AMD推出基于RV670核心的系列产品获得市场好评。其系列中的顶级型号Radeon HD 3870的3D性能较上一代旗舰R600核心的Radeon HD 2900 XT有较大提升。不过从我们以往的测试来看，Radeon HD 3870的3D性能并不如ONVIDIA的顶级产品GeForce 8800 Ultra。基于R680核心的新旗舰产品——Radeon HD 3870×2，就是在这样背景下诞生，AMD意在靠它重夺显卡旗舰霸主。
