微管类药物在转移、耐药的癌症治疗中发挥重要的作用。然而,临床上出现微管类药物耐药现象,需要研究者研发具有新作用模式、新机制的分子来克服耐药问题。多靶点药物分子策略是近年来针对复杂性疾病及耐药问题发展起来的一种药物分子设...微管类药物在转移、耐药的癌症治疗中发挥重要的作用。然而,临床上出现微管类药物耐药现象,需要研究者研发具有新作用模式、新机制的分子来克服耐药问题。多靶点药物分子策略是近年来针对复杂性疾病及耐药问题发展起来的一种药物分子设计策略。本文在替巴尼布林基础上,结合课题组前期研究,开展微管蛋白(tubulin)及原癌基因酪氨酸蛋白激酶Src(proto-oncogene tyrosine protein kinase Src,c-Src)双靶点抑制剂研究。从增加饱和碳原子比率(fraction of sp^(3)-hybridized carbon atoms,Fsp^(3))角度出发,合成16个目标化合物,并通过核磁共振氢谱、碳谱和质谱对其结构进行表征;采用噻唑蓝(MTT)法评价了化合物对宫颈癌细胞(HeLa)和肝癌细胞(HepG2)两种细胞株的生长抑制活性,并通过免疫荧光和流式细胞术评价化合物22对微管和细胞周期的影响。结果显示2-咪唑啉酮不适合替代替巴尼布林中的吡啶结构,化合物22对HeLa和HepG2的半数生长抑制浓度分别为45和51 nmol·L^(-1),并且能够破坏微管结构,将细胞周期阻滞于G2/M期。化合物22可作为先导物开展进一步深入研究。展开更多
Background:Expression of mRNA is widely regulated by N6-methyladenosine(m6A).An increasing number of studies have shown that m6A methylation,facilitated by methyltransferase 3(METTL3),is crucial in the progression of ...Background:Expression of mRNA is widely regulated by N6-methyladenosine(m6A).An increasing number of studies have shown that m6A methylation,facilitated by methyltransferase 3(METTL3),is crucial in the progression of tumors.Previous reports have indicated the involvement of both METTL3 and c-Src kinase in the evolution of liver cancer.However,the potential connection between c-Src and the METTL3-mediated mechanism in liver cancer progression remains elusive.Methods:The correlation expression between c-Src and METTL3 between liver cancer patients and the control group was analyzed using the TCGA database,and was further demonstrated by Western blot and RT-qPCR.The functional roles of c-Src in METTL3-regulated liver cancer progression were investigated by cell proliferation assays and colony formation assays.The regulatory mechanism of METTL3 in c-Src expression was accessed by RNA-immunoprecipitation(RIP)-qPCR.Results:We demonstrated that c-Src kinase promoted liver cancer development,and the expression of SRC(encodes c-Src kinase)was positively correlated with METTL3 in liver cancer cases.We showed that SRC mRNA could be m6A-modified,and METTL3 regulated the transcription of SRC mRNA through interferon regulatory factor 1(IRF1).We revealed that IRF1,the expression of which was positively regulated byMETTL3,was a novel transcription factor of c-Src.Lastly,The pro-proliferative effect of METTL3 on hepatocellular carcinoma was mechanistically linked to IRF1/c-Src axis activation,as evidenced by our experimental data.Conclusion:Results suggested that the METTL3/IRF1/c-Src axis played potential oncogenic roles in liver cancer development and the axis may be a promising therapeutic target in the disease.展开更多
本文旨在研究c-SRC蛋白对人宫颈癌HeLa细胞的活性及对磷酸化的信号转导与转录激活子-3(phosphorylated signal transducer and activator of transcription-3,p-STAT3)表达的影响。人宫颈癌HeLa细胞转染c-SRC RNA干涉质粒后,分别用RT-PC...本文旨在研究c-SRC蛋白对人宫颈癌HeLa细胞的活性及对磷酸化的信号转导与转录激活子-3(phosphorylated signal transducer and activator of transcription-3,p-STAT3)表达的影响。人宫颈癌HeLa细胞转染c-SRC RNA干涉质粒后,分别用RT-PCR和Westernblot检测细胞内c-SRC mRNA和蛋白的表达;用MTT比色法观察c-SRC敲减后细胞的活性;用流式细胞仪检测细胞周期;同时检测细胞内p-STAT3的表达情况。转染c-SRC RNA干涉质粒后,HeLa细胞内c-SRC mRNA和蛋白的表达显著降低;在转染c-SRC RNA干涉质粒24、48、72及96h后,细胞活性分别下降了23.1%、29.3%、38.6%和45.0%(均P<0.05)。转染c-SRC RNA干涉质粒24、48、72及96h后,HeLa细胞S期细胞数分别下降了5.6%、10.0%、15.2%和19.9%(均P<0.05)。敲减c-SRC后,细胞内p-STAT3的含量也显著下降。与对照组相比,STAT3抑制剂Piceatannol处理细胞24、48、72、96h后,细胞活性分别下降了23.8%、29.7%、37.3%和45.4%(均P<0.05),而Piceatannol预处理细胞后再用重组人c-SRC蛋白处理增加细胞内c-SRC蛋白的含量,细胞活性未见明显增加。以上结果表明,c-SRC敲减后抑制HeLa细胞的活性可能与其抑制STAT3蛋白磷酸化相关。展开更多
背景:原癌基因c-Src在调节高血压等心血管系统疾病中起着重要的作用,目前尚未看到有关运动干预影响主动脉血管内皮细胞c-Src的表达和活来调节高血压的研究。目的:观察有氧运动对自发性高血压大鼠主动脉血管内皮细胞c-Src m RNA表达和c-...背景:原癌基因c-Src在调节高血压等心血管系统疾病中起着重要的作用,目前尚未看到有关运动干预影响主动脉血管内皮细胞c-Src的表达和活来调节高血压的研究。目的:观察有氧运动对自发性高血压大鼠主动脉血管内皮细胞c-Src m RNA表达和c-Src活性的影响。方法:8只雄性Wistar大鼠作为正常对照组,16只自发性高血压大鼠随机分为自发性高血压组8只,自发性高血压运动组8只。自发性高血压运动组每天进行90 min无负重有氧游泳运动,6 d/周,共8周;正常对照组和自发性高血压组大鼠不做游泳运动。每周测定大鼠血压1次,8周后,测定各组大鼠主动脉血管内皮细胞c-Src m RNA表达和c-Src活性。结果与结论:与自发性高血压组相比,自发性高血压运动组血压明显降低,主动脉血管内皮细胞c-Src m RNA表达和c-Src活性明显升高。与正常对照组相比,自发性高血压运动组大鼠主动脉血管内皮细胞c-Src活性和c-Src m RNA表达高于正常对照组和自发性高血压组(P<0.01)。结果提示有氧运动可以促进自发性高血压大鼠主动脉血管内皮细胞c-Src活性和c-Src m RNA表达的增加。展开更多
文摘微管类药物在转移、耐药的癌症治疗中发挥重要的作用。然而,临床上出现微管类药物耐药现象,需要研究者研发具有新作用模式、新机制的分子来克服耐药问题。多靶点药物分子策略是近年来针对复杂性疾病及耐药问题发展起来的一种药物分子设计策略。本文在替巴尼布林基础上,结合课题组前期研究,开展微管蛋白(tubulin)及原癌基因酪氨酸蛋白激酶Src(proto-oncogene tyrosine protein kinase Src,c-Src)双靶点抑制剂研究。从增加饱和碳原子比率(fraction of sp^(3)-hybridized carbon atoms,Fsp^(3))角度出发,合成16个目标化合物,并通过核磁共振氢谱、碳谱和质谱对其结构进行表征;采用噻唑蓝(MTT)法评价了化合物对宫颈癌细胞(HeLa)和肝癌细胞(HepG2)两种细胞株的生长抑制活性,并通过免疫荧光和流式细胞术评价化合物22对微管和细胞周期的影响。结果显示2-咪唑啉酮不适合替代替巴尼布林中的吡啶结构,化合物22对HeLa和HepG2的半数生长抑制浓度分别为45和51 nmol·L^(-1),并且能够破坏微管结构,将细胞周期阻滞于G2/M期。化合物22可作为先导物开展进一步深入研究。
基金supported by Natural Science Foundation of Hunan Province of China(project No.2022JJ40413)Outstanding Youth Project of Hunan Provincial Department of Education(project No.22B0814)+1 种基金Regional Consolidated Foundation ofHunan Province of China(project No.2023JJ50065)Natural Science Foundation of Hunan Province of China(project No.2023JJ50412).
文摘Background:Expression of mRNA is widely regulated by N6-methyladenosine(m6A).An increasing number of studies have shown that m6A methylation,facilitated by methyltransferase 3(METTL3),is crucial in the progression of tumors.Previous reports have indicated the involvement of both METTL3 and c-Src kinase in the evolution of liver cancer.However,the potential connection between c-Src and the METTL3-mediated mechanism in liver cancer progression remains elusive.Methods:The correlation expression between c-Src and METTL3 between liver cancer patients and the control group was analyzed using the TCGA database,and was further demonstrated by Western blot and RT-qPCR.The functional roles of c-Src in METTL3-regulated liver cancer progression were investigated by cell proliferation assays and colony formation assays.The regulatory mechanism of METTL3 in c-Src expression was accessed by RNA-immunoprecipitation(RIP)-qPCR.Results:We demonstrated that c-Src kinase promoted liver cancer development,and the expression of SRC(encodes c-Src kinase)was positively correlated with METTL3 in liver cancer cases.We showed that SRC mRNA could be m6A-modified,and METTL3 regulated the transcription of SRC mRNA through interferon regulatory factor 1(IRF1).We revealed that IRF1,the expression of which was positively regulated byMETTL3,was a novel transcription factor of c-Src.Lastly,The pro-proliferative effect of METTL3 on hepatocellular carcinoma was mechanistically linked to IRF1/c-Src axis activation,as evidenced by our experimental data.Conclusion:Results suggested that the METTL3/IRF1/c-Src axis played potential oncogenic roles in liver cancer development and the axis may be a promising therapeutic target in the disease.
基金supported by the National Natural Science Foundation of China(No.30360032)the Science Foundation of Educational Committee of Jiangxi Province+3 种基金China(No.GJJ09444GJJ09111)the Scientific Research Foundation for the Scholars of Ph.D.Degree of Nanchang University and Scientific Research Foundation of Nanchang UniversityChina
文摘本文旨在研究c-SRC蛋白对人宫颈癌HeLa细胞的活性及对磷酸化的信号转导与转录激活子-3(phosphorylated signal transducer and activator of transcription-3,p-STAT3)表达的影响。人宫颈癌HeLa细胞转染c-SRC RNA干涉质粒后,分别用RT-PCR和Westernblot检测细胞内c-SRC mRNA和蛋白的表达;用MTT比色法观察c-SRC敲减后细胞的活性;用流式细胞仪检测细胞周期;同时检测细胞内p-STAT3的表达情况。转染c-SRC RNA干涉质粒后,HeLa细胞内c-SRC mRNA和蛋白的表达显著降低;在转染c-SRC RNA干涉质粒24、48、72及96h后,细胞活性分别下降了23.1%、29.3%、38.6%和45.0%(均P<0.05)。转染c-SRC RNA干涉质粒24、48、72及96h后,HeLa细胞S期细胞数分别下降了5.6%、10.0%、15.2%和19.9%(均P<0.05)。敲减c-SRC后,细胞内p-STAT3的含量也显著下降。与对照组相比,STAT3抑制剂Piceatannol处理细胞24、48、72、96h后,细胞活性分别下降了23.8%、29.7%、37.3%和45.4%(均P<0.05),而Piceatannol预处理细胞后再用重组人c-SRC蛋白处理增加细胞内c-SRC蛋白的含量,细胞活性未见明显增加。以上结果表明,c-SRC敲减后抑制HeLa细胞的活性可能与其抑制STAT3蛋白磷酸化相关。
文摘背景:原癌基因c-Src在调节高血压等心血管系统疾病中起着重要的作用,目前尚未看到有关运动干预影响主动脉血管内皮细胞c-Src的表达和活来调节高血压的研究。目的:观察有氧运动对自发性高血压大鼠主动脉血管内皮细胞c-Src m RNA表达和c-Src活性的影响。方法:8只雄性Wistar大鼠作为正常对照组,16只自发性高血压大鼠随机分为自发性高血压组8只,自发性高血压运动组8只。自发性高血压运动组每天进行90 min无负重有氧游泳运动,6 d/周,共8周;正常对照组和自发性高血压组大鼠不做游泳运动。每周测定大鼠血压1次,8周后,测定各组大鼠主动脉血管内皮细胞c-Src m RNA表达和c-Src活性。结果与结论:与自发性高血压组相比,自发性高血压运动组血压明显降低,主动脉血管内皮细胞c-Src m RNA表达和c-Src活性明显升高。与正常对照组相比,自发性高血压运动组大鼠主动脉血管内皮细胞c-Src活性和c-Src m RNA表达高于正常对照组和自发性高血压组(P<0.01)。结果提示有氧运动可以促进自发性高血压大鼠主动脉血管内皮细胞c-Src活性和c-Src m RNA表达的增加。
