A series of novel E-ring modified evodiamine derivatives were designed and synthesized as antitumor agents. Their capacity to interfere with the catalytic activity of topoisomerase Ⅰ and Ⅱ was evaluated by the relax...A series of novel E-ring modified evodiamine derivatives were designed and synthesized as antitumor agents. Their capacity to interfere with the catalytic activity of topoisomerase Ⅰ and Ⅱ was evaluated by the relaxation assay. In vitro antitumor activity results revealed that compound 12 showed good antitumor activity with a broad spectrum. Its binding modes with topoisomerase Ⅰ and Ⅱ were clarified by molecular docking.展开更多
背景与目的:卵巢癌是妇科恶性肿瘤中发病率排名第3、致死率排名第1的疾病。卵巢癌的预后很差,这是由于大部分患者确诊时已处于晚期并且对铂类药物化疗易耐药,异常的DNA修复是导致铂类耐药的重要原因,针对性地干扰DNA损伤修复相关分子可...背景与目的:卵巢癌是妇科恶性肿瘤中发病率排名第3、致死率排名第1的疾病。卵巢癌的预后很差,这是由于大部分患者确诊时已处于晚期并且对铂类药物化疗易耐药,异常的DNA修复是导致铂类耐药的重要原因,针对性地干扰DNA损伤修复相关分子可能是提高铂类药物化疗敏感性的新手段。Ku86是参与非同源末端连接(non-homologous end joining,NHEJ)过程的一个关键分子,能够有效地修复DNA双链断裂(DNA double-strand break,DSB)。方法:使用免疫组织化学染色和细胞免疫荧光法检测Ku86在卵巢癌组织和细胞中的定位。使用RNAi技术下调Ku86,用顺铂处理后使用流式细胞术检测凋亡,使用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测IC_(50)。使用蛋白质印迹法(Western blot)分别检测并分析Ku86与拓扑异构酶Ⅰ(topoisomeraseⅠ,TOP1)及COP9信号复合体的第5种成分(the fifth component of the COP9 signalosome,COPS5)的关系。结果:下调Ku86可以减少由于顺铂引起的细胞凋亡,降低卵巢癌细胞对于顺铂的敏感性。Ku86对顺铂在卵巢癌药物敏感性中的这种影响可能是通过调节TOP1和COPS5发挥作用的。结论:TOP1和COPS5都是与DNA损伤修复有关并能影响铂类药物化疗敏感性的重要分子,下调Ku86会使TOP1的表达增高和COPS5的表达减少。卵巢癌中与铂类药物敏感性有关的生物标志物之间有相关性,靶向Ku86的治疗可能是提高卵巢癌药物敏感性的有效策略。展开更多
OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities.The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in v...OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities.The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS The antitumor effect of lapachol was firstly evaluated in the C6 glioma model in Wistar rats.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phenazinemethosulfate(PMS)assay,hoechst 33358 staining,annexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomerase I(TOP I)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322DNA relaxation assays and molecular docking.TOPⅠand TOPⅡexpression levels in C6 cells were also determined.RESULTS High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).It was showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cel s in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠ and Ⅱ.Lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡin molecular docking study.Also,lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠ and TOPⅡ activities,as wel as TOPⅡ expression.展开更多
基金supported by National Natural Science Foundation of China(Nos.81222044,81373278)Key Project of Science and Technology of Shanghai(No.11431920402)+2 种基金the National Basic Research Program of China(No.2014CB541800)Shanghai Rising Star Program(No.12QH1402600)Shanghai Municipal Health Bureau(No.XYQ2011038)
文摘A series of novel E-ring modified evodiamine derivatives were designed and synthesized as antitumor agents. Their capacity to interfere with the catalytic activity of topoisomerase Ⅰ and Ⅱ was evaluated by the relaxation assay. In vitro antitumor activity results revealed that compound 12 showed good antitumor activity with a broad spectrum. Its binding modes with topoisomerase Ⅰ and Ⅱ were clarified by molecular docking.
文摘背景与目的:卵巢癌是妇科恶性肿瘤中发病率排名第3、致死率排名第1的疾病。卵巢癌的预后很差,这是由于大部分患者确诊时已处于晚期并且对铂类药物化疗易耐药,异常的DNA修复是导致铂类耐药的重要原因,针对性地干扰DNA损伤修复相关分子可能是提高铂类药物化疗敏感性的新手段。Ku86是参与非同源末端连接(non-homologous end joining,NHEJ)过程的一个关键分子,能够有效地修复DNA双链断裂(DNA double-strand break,DSB)。方法:使用免疫组织化学染色和细胞免疫荧光法检测Ku86在卵巢癌组织和细胞中的定位。使用RNAi技术下调Ku86,用顺铂处理后使用流式细胞术检测凋亡,使用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测IC_(50)。使用蛋白质印迹法(Western blot)分别检测并分析Ku86与拓扑异构酶Ⅰ(topoisomeraseⅠ,TOP1)及COP9信号复合体的第5种成分(the fifth component of the COP9 signalosome,COPS5)的关系。结果:下调Ku86可以减少由于顺铂引起的细胞凋亡,降低卵巢癌细胞对于顺铂的敏感性。Ku86对顺铂在卵巢癌药物敏感性中的这种影响可能是通过调节TOP1和COPS5发挥作用的。结论:TOP1和COPS5都是与DNA损伤修复有关并能影响铂类药物化疗敏感性的重要分子,下调Ku86会使TOP1的表达增高和COPS5的表达减少。卵巢癌中与铂类药物敏感性有关的生物标志物之间有相关性,靶向Ku86的治疗可能是提高卵巢癌药物敏感性的有效策略。
文摘OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities.The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS The antitumor effect of lapachol was firstly evaluated in the C6 glioma model in Wistar rats.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phenazinemethosulfate(PMS)assay,hoechst 33358 staining,annexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomerase I(TOP I)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322DNA relaxation assays and molecular docking.TOPⅠand TOPⅡexpression levels in C6 cells were also determined.RESULTS High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).It was showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cel s in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠ and Ⅱ.Lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡin molecular docking study.Also,lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠ and TOPⅡ activities,as wel as TOPⅡ expression.
