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
目的比较通辽地区不同来源奇异变形杆菌对小鼠致病力及其毒力基因携带情况。方法取4株(SPM、GPM、BPM-1和BPM-2)不同来源的奇异变形杆菌,进行生化鉴定、药物敏感性测试及动物致病性试验,对4株菌株的8种毒力基因(ureC、zapA、mrpA、ucaA...目的比较通辽地区不同来源奇异变形杆菌对小鼠致病力及其毒力基因携带情况。方法取4株(SPM、GPM、BPM-1和BPM-2)不同来源的奇异变形杆菌,进行生化鉴定、药物敏感性测试及动物致病性试验,对4株菌株的8种毒力基因(ureC、zapA、mrpA、ucaA、rsbA、pmfA、atfA、atfC)进行PCR检测,对其16S r RNA基因序列进行同源性和系统进化树分析。结果 4株菌株均被鉴定为奇异变形杆菌;对4株病原菌均高度敏感的药物有丁胺卡那、美罗培南、头孢他啶、头孢噻肟、头孢吡肟、他巴克坦和氨曲南7种,均不敏感的药物有庆大霉素、亚胺培南、头孢唑啉、氨苄西林、克拉维酸、黏菌素、磺胺甲恶唑、氯霉素、环丙沙星和四环素10种;4株菌株的半数致死量(LD50)按BPM-2、BPM-1、SPM、GPM依次降低,鹅源的毒力最高;奇异变形杆菌具有不同的毒力基因谱,8种毒力基因在菌株SPM、GPM、BPM-2中均被检出,菌株BPM-2未检出ucaA基因。BPM-2和BPM-1处于同一分支,但分处于不同的末支;SPM和GPM分处于不同的小分支。结论不同来源的奇异变形杆菌对小鼠的致病力存在较小的差异,且对小鼠有致病力的基因表型与该毒力基因的分布可能存在一定的相关性。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.
文摘目的比较通辽地区不同来源奇异变形杆菌对小鼠致病力及其毒力基因携带情况。方法取4株(SPM、GPM、BPM-1和BPM-2)不同来源的奇异变形杆菌,进行生化鉴定、药物敏感性测试及动物致病性试验,对4株菌株的8种毒力基因(ureC、zapA、mrpA、ucaA、rsbA、pmfA、atfA、atfC)进行PCR检测,对其16S r RNA基因序列进行同源性和系统进化树分析。结果 4株菌株均被鉴定为奇异变形杆菌;对4株病原菌均高度敏感的药物有丁胺卡那、美罗培南、头孢他啶、头孢噻肟、头孢吡肟、他巴克坦和氨曲南7种,均不敏感的药物有庆大霉素、亚胺培南、头孢唑啉、氨苄西林、克拉维酸、黏菌素、磺胺甲恶唑、氯霉素、环丙沙星和四环素10种;4株菌株的半数致死量(LD50)按BPM-2、BPM-1、SPM、GPM依次降低,鹅源的毒力最高;奇异变形杆菌具有不同的毒力基因谱,8种毒力基因在菌株SPM、GPM、BPM-2中均被检出,菌株BPM-2未检出ucaA基因。BPM-2和BPM-1处于同一分支,但分处于不同的末支;SPM和GPM分处于不同的小分支。结论不同来源的奇异变形杆菌对小鼠的致病力存在较小的差异,且对小鼠有致病力的基因表型与该毒力基因的分布可能存在一定的相关性。
