siRNA沉默MAPK p42对肝癌SMMC7721增殖和凋亡的影响

目的应用siRNA沉默SMMC7721细胞MAPK p42,并观察其对增殖和凋亡的影响。方法应用SilencerTMsiRNA构建试剂盒合成2条MAPK p42siRNAs和一条随机对照siRNAs,用脂质体LipofectaminTM 2000将其转染入肝癌SMMC-7721细胞株。应用Western blot检测MAPK p42表达,MTT法检测细胞增殖情况,流式细胞仪分析细胞周期及其细胞凋亡。结果与对照相比,MAPK p42siRNA明显抑制MAPK p42表达。MAPK p42siRNA抑制了SMMC-7721细胞的生长,将细胞阻滞于细胞周期S期,进而诱导了细胞凋亡的发生。结论 MAPK p42靶向siRNA可能为肝癌基因治疗的重要手段。     

β淀粉样前体蛋白裂解酶特异性小干扰RNA真核表达载体的构建和鉴定

目的构建β淀粉样前体蛋白裂解酶(BACE)特异性小干扰RNA真核表达载体。方法用PCR扩增BACE特异性小干扰RNA,转入带有增强型绿荧光蛋白(EGFP)和启动子U6的pUC19质粒,再将重组基因片段导入逆转录病毒真核表达载体pLXSN中,通过限制性酶切对该重组表达载体进行鉴定。结果成功构建了BACE真核表达载体pLXSN/EGFP-U6-siBACE。结论pLXSN/EGFP-U6-siBACE载体的成功构建对进一步利用基因干扰技术治疗阿尔茨海默病的研究奠定了重要的基础。     

CONSTRUCTION AND IDENTIFICATION OF EUKARYOTIC VECTOR EXPRESSING SIRNA SPECIFIC FOR BACE

TheAlzheimer sdisease(AD)isaseriousde generativediseaseandmanifestedthedepositionof amyloid peptide(Aβ)intosenileplaquesintheex tracellularspaceandthehyperphosphorylationoftau proteincausingneurofibrillarytangles[1].RNAinterference(RNAi)isanancientevolu …     

siRNA directed against TrkA sensitizes human pancreatic cancer cells to apoptosis induced by gemcitabine through an inactivation of PI3K/Akt-dependent pathway

Pancreatic cancer is a lethal disease.Despite sig-nificant advances in diagnosis,staging,and surgicalmanagement of the disease,less than10%of pa-tients survive for more than one year fromthe ti meof diagnosis[1-2].The largely refractory response tochemotherapy is a common feature of this disease,and eventhe principal therapeutic agent,the nucleo-side analogue gemcitabine(2′,2′-difluorodeoxycyt-idine),only marginally affects clinical outcomes[3].Therefore,further understanding of the chemores…