OBJECTIVE The present study was designed to investigate anticancer effect of zeylenone(Zey)on K562 cells derived from chronic myelogenous leukemia(CML)both in vitro and in vivo,followed by exploring the underlying mec...OBJECTIVE The present study was designed to investigate anticancer effect of zeylenone(Zey)on K562 cells derived from chronic myelogenous leukemia(CML)both in vitro and in vivo,followed by exploring the underlying mechanisms.METHODS Initially,the effects of Zey on cel viability,proliferation,and apoptosis were measured in K562 cells by MTT,soft agar assay,AO/EB staining,hoechst 33258 staining and flow cytometric analysis after they were treated with Zey for indicated time,the involving signaling pathways were then investigated by JC-1,real-time quantitative polymerase chain reaction(RT-q PCR),Western blotting and immunofluorescence analysis.Furthermore,the in vivo anti-tumoractivity of Zey was assessed with nude xenografts and the involving mechanism was confirmed by immunohistochemical(IHC)and histopathological analysis.RESULTS We identified that Zey dose-dependently decreased cell viability,colony formation and expression of Proliferating Cell Nuclear Antigen(PCNA),and significantly induced K562 cell apoptosis via regulating Bcl-2 family members,decreasing mitochondrial transmembrane potential,and activating caspase-3,caspase-9,and caspase-8(P<0.05 or P<0.01).Further study revealed that Zey significantly inhibited phosphorylation of Jak2 and Src and downregulated their downstream proteins,including stat3,PI3K/AKT/m TOR,and ERK1/2 signaling pathways(P<0.05 or P<0.01).Zey also suppressed tumor growth with low toxicity in mouse xenograft model of K562cells through decreasing expression of Jak2 and Src.CONCLUSION Our data demonstrated that Zey substantially suppressed K562 cells both in vitro and in vivo through Jak2 and Src pathways.These findings suggest the potential of Zey as an effective anticancer agent in CML treatment.展开更多
本研究观察程序性细胞死亡5基因(programmed cell death 5,PDCD5)重组腺病毒转染K562细胞后对化疗药物依托泊甙的增敏作用。利用AdMaxTM腺病毒载体包装系统,通过同源重组方法构建Ad-PDCD5重组腺病毒及对照腺病毒Ad-null及Ad-eGFP;用不...本研究观察程序性细胞死亡5基因(programmed cell death 5,PDCD5)重组腺病毒转染K562细胞后对化疗药物依托泊甙的增敏作用。利用AdMaxTM腺病毒载体包装系统,通过同源重组方法构建Ad-PDCD5重组腺病毒及对照腺病毒Ad-null及Ad-eGFP;用不同感染复数将Ad-eGFP、Ad-null或Ad-PDCD5转染人白血病细胞系,实时定量PCR检测PDCD5mRNA的相对表达水平;利用MTT法及Annexin-V-FITC/PI双染色流式细胞术观察依托泊甙对转染后K562细胞增殖与凋亡的影响。结果表明:Ad-eGFP腺病毒对白血病细胞系K562、Jurkat及CEM的转染效率可达60%-86%。Ad-PDCD5重组腺病毒能梯度增加K562细胞PDCD5 mRNA的相对表达水平,腺病毒介导的PDCD5基因转移促进依托泊甙诱导的K562细胞凋亡。结论:PDCD5重组腺病毒可能成为化疗药物的增敏剂。展开更多
基金The project supported by PUMC Youth Fund(3332015047)Fundamental Research Funds for the Central Universities,Beijing Key Laboratory of Innovative Drug Discovery of Traditional Chinese Medicine(Natural Medicine)and Translational Medicine,Institute of Medical Plant Development,Peking Union Medical College and Chinese Academy of Medical Sciencesby the National Science and Technology Major Project and Scientific Researchers Aiding Enterprise Item(2012ZX09301-002-001-026 and 2012ZX09501001-004)from the Ministry of Science and Technology of China
文摘OBJECTIVE The present study was designed to investigate anticancer effect of zeylenone(Zey)on K562 cells derived from chronic myelogenous leukemia(CML)both in vitro and in vivo,followed by exploring the underlying mechanisms.METHODS Initially,the effects of Zey on cel viability,proliferation,and apoptosis were measured in K562 cells by MTT,soft agar assay,AO/EB staining,hoechst 33258 staining and flow cytometric analysis after they were treated with Zey for indicated time,the involving signaling pathways were then investigated by JC-1,real-time quantitative polymerase chain reaction(RT-q PCR),Western blotting and immunofluorescence analysis.Furthermore,the in vivo anti-tumoractivity of Zey was assessed with nude xenografts and the involving mechanism was confirmed by immunohistochemical(IHC)and histopathological analysis.RESULTS We identified that Zey dose-dependently decreased cell viability,colony formation and expression of Proliferating Cell Nuclear Antigen(PCNA),and significantly induced K562 cell apoptosis via regulating Bcl-2 family members,decreasing mitochondrial transmembrane potential,and activating caspase-3,caspase-9,and caspase-8(P<0.05 or P<0.01).Further study revealed that Zey significantly inhibited phosphorylation of Jak2 and Src and downregulated their downstream proteins,including stat3,PI3K/AKT/m TOR,and ERK1/2 signaling pathways(P<0.05 or P<0.01).Zey also suppressed tumor growth with low toxicity in mouse xenograft model of K562cells through decreasing expression of Jak2 and Src.CONCLUSION Our data demonstrated that Zey substantially suppressed K562 cells both in vitro and in vivo through Jak2 and Src pathways.These findings suggest the potential of Zey as an effective anticancer agent in CML treatment.
文摘本研究观察程序性细胞死亡5基因(programmed cell death 5,PDCD5)重组腺病毒转染K562细胞后对化疗药物依托泊甙的增敏作用。利用AdMaxTM腺病毒载体包装系统,通过同源重组方法构建Ad-PDCD5重组腺病毒及对照腺病毒Ad-null及Ad-eGFP;用不同感染复数将Ad-eGFP、Ad-null或Ad-PDCD5转染人白血病细胞系,实时定量PCR检测PDCD5mRNA的相对表达水平;利用MTT法及Annexin-V-FITC/PI双染色流式细胞术观察依托泊甙对转染后K562细胞增殖与凋亡的影响。结果表明:Ad-eGFP腺病毒对白血病细胞系K562、Jurkat及CEM的转染效率可达60%-86%。Ad-PDCD5重组腺病毒能梯度增加K562细胞PDCD5 mRNA的相对表达水平,腺病毒介导的PDCD5基因转移促进依托泊甙诱导的K562细胞凋亡。结论:PDCD5重组腺病毒可能成为化疗药物的增敏剂。
