Dvl1及其结构缺失突变体ΔDIX(dsh and axin)和ΔDEP(dsh,Egl-10 and pleckstrin)腺病毒载体的构建并验证其在MSCs(mesenchymal stem cells)中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中...Dvl1及其结构缺失突变体ΔDIX(dsh and axin)和ΔDEP(dsh,Egl-10 and pleckstrin)腺病毒载体的构建并验证其在MSCs(mesenchymal stem cells)中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中与pAdEasy-1骨架质粒同源重组,获得重组腺病毒载体,在QBI-293A细胞中包装及扩增,实时荧光定量PCR和Western bolt验证Dvl1在MSCs中的表达情况。经PCR、PacⅠ单酶切鉴定及测序分析,成功构建Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,目的基因序列与GenBank报道一致,并选出150 MOI为最适感染MSCs的感染复数,成功构建了Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,并获得高滴度的病毒子,qPCR和Western blot证实Dvl1在MSCs中高表达并增加了β-catenin在细胞中的累积,为进一步研究Dvl1在MSCs迁移过程的作用奠定了基础。展开更多
Objective:To investigate the antioxidant,antimicrobial,and anticancer effects of Achillea filipendulina.Methods:Phytochemical content was analyzed by GC-MS.Antioxidant activity was assayed by DPPH,total antioxidant st...Objective:To investigate the antioxidant,antimicrobial,and anticancer effects of Achillea filipendulina.Methods:Phytochemical content was analyzed by GC-MS.Antioxidant activity was assayed by DPPH,total antioxidant status,total oxidant status,and iron reduction tests.Antimicrobial activity was assessed using the MIC assay.Anticancer activity was detected via MTT assay,followed by analysis of the apoptotic process using annexin V and caspase 3/7.miR-519d and Dvl-1 expression were assessed by RT-PCR,and molecular docking analysis was also performed.Results:The ethanol extract of Achillea filipendulina showed the strongest anticancer effect.A significant increase in miR-519d expression was observed in HT29 cells,while a decrease in Dvl-1 expression was noted.DPPH,FRAP,total antioxidant status,and total oxidant assays showed high antioxidant activity.The hexane and ethanol extracts were effective against Streptococcus mutans(MIC<50μg/mL),while the ethanol extract was effective against Enterococcus faecalis and Candida albicans(MIC<50μg/mL).Molecular docking studies also confirmed that ethanol extracts could act on the Wnt pathway.GC-MS analysis showed high level of eucalyptol and borneol content in the extract.Conclusions:Ethanol extract of Achillea filipendulina has anticancer effect and can be used in colon cancer treatments.展开更多
文摘Dvl1及其结构缺失突变体ΔDIX(dsh and axin)和ΔDEP(dsh,Egl-10 and pleckstrin)腺病毒载体的构建并验证其在MSCs(mesenchymal stem cells)中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中与pAdEasy-1骨架质粒同源重组,获得重组腺病毒载体,在QBI-293A细胞中包装及扩增,实时荧光定量PCR和Western bolt验证Dvl1在MSCs中的表达情况。经PCR、PacⅠ单酶切鉴定及测序分析,成功构建Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,目的基因序列与GenBank报道一致,并选出150 MOI为最适感染MSCs的感染复数,成功构建了Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,并获得高滴度的病毒子,qPCR和Western blot证实Dvl1在MSCs中高表达并增加了β-catenin在细胞中的累积,为进一步研究Dvl1在MSCs迁移过程的作用奠定了基础。
基金supported by the TUBITAK 2209-A University Student Project(Project Number:1919B012111858).
文摘Objective:To investigate the antioxidant,antimicrobial,and anticancer effects of Achillea filipendulina.Methods:Phytochemical content was analyzed by GC-MS.Antioxidant activity was assayed by DPPH,total antioxidant status,total oxidant status,and iron reduction tests.Antimicrobial activity was assessed using the MIC assay.Anticancer activity was detected via MTT assay,followed by analysis of the apoptotic process using annexin V and caspase 3/7.miR-519d and Dvl-1 expression were assessed by RT-PCR,and molecular docking analysis was also performed.Results:The ethanol extract of Achillea filipendulina showed the strongest anticancer effect.A significant increase in miR-519d expression was observed in HT29 cells,while a decrease in Dvl-1 expression was noted.DPPH,FRAP,total antioxidant status,and total oxidant assays showed high antioxidant activity.The hexane and ethanol extracts were effective against Streptococcus mutans(MIC<50μg/mL),while the ethanol extract was effective against Enterococcus faecalis and Candida albicans(MIC<50μg/mL).Molecular docking studies also confirmed that ethanol extracts could act on the Wnt pathway.GC-MS analysis showed high level of eucalyptol and borneol content in the extract.Conclusions:Ethanol extract of Achillea filipendulina has anticancer effect and can be used in colon cancer treatments.
