AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric carcinogenesis.METHODS:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-ti...AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric carcinogenesis.METHODS:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-time quantitative RT-PCR in gastric tumors and their matched non-neoplastic gastric tissue samples.RESULTS:Authors observed a significant reduction of 14-3-3εprotein expression in gastric cancer(GC)samples compared to their matched non-neoplastic tissue.Reduced levels of 14-3-3εwere also associated with diffuse-type GC and early-onset of this pathology.Our data suggest that reduced 14-3-3εmay have a role in gastric carcinogenesis process.CONCLUSION:Our results reveal that the reduced 14-3-3εexpression in GC and investigation of 14-3-3ε interaction partners may help to elucidate the carcino-genesis process.展开更多
文摘目的探讨寨卡病毒(Zika virus,ZIKV)逃避子宫成纤维细胞先天免疫的分子机制。方法使用ZIKV感染子宫成纤维细胞后,检测YWHAE-RIG-I-MAVS-IFN-β通路关键分子的表达情况。通过实时荧光定量PCR检测YWHAE mRNA的表达情况。通过高通量测序检测ZIKV感染子宫成纤维细胞后的miRNA表达谱。通过荧光素酶报告实验确定调控YWHAE的miRNA。结果ZIKV感染子宫成纤维细胞后,IFN-β的分泌和IFN-β的启动子活性水平下降(P<0.05),YWHAE、p-IRF3S396的表达水平下降,MAVS与RIG-I、RIG-I与YWHAE的相互作用水平下降。但是,YWHAE启动子活性没有显著改变(P>0.05),而YWHAE mRNA水平下降(P<0.05)。通过高通量测序发现ZIKV感染后子宫成纤维细胞miRNA的表达发生改变。其中hsamiR-15a-5p、hsa-miR-6780b-3p、hsa-miR-6845-3p、hsa-miR-16-5p、hsa-miR-590-3p、hsa-miR-575、hsa-miR-4674、hsamiR-371a-5p、hsa-miR-4642、hsa-miR-4676-5p的表达差异在5倍以上。子宫成纤维细胞中过表达上述差异miRNA后,miR-545-3p能够降低YWHAE mRNA的表达水平(P<0.05)。敲低miR-545-3p后,YWHAE m RNA和蛋白的表达水平上升(P<0.05)。此外,miR-545-3p靶向YWHAE的3端非编码区(P<0.05)。ZIKV感染子宫成纤维细胞的同时过表达miR-545-3p后,YWHAE mRNA水平显著下降(P<0.05)、IFN-β分泌水平显著下降(P<0.05)、ZIKV复制水平均显著上升(P<0.05);但是,ZIKV感染子宫成纤维细胞的同时敲低miR-545-3p后,YWHAE m RNA水平显著上升(P<0.05)、IFN-β分泌水平显著上升(P<0.05)、ZIKV复制水平均显著下降(P<0.05)。结论ZIKV通过miR-545-3p靶向YWHAE促进了自身的复制。
基金Supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico(CNPqChammas R,Smith MAC and Burbano RR)+1 种基金Fundao de Amparoà Pesquisa do Estado de So Paulo (FAPESPLeal MF and Calcagno DQ)
文摘AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric carcinogenesis.METHODS:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-time quantitative RT-PCR in gastric tumors and their matched non-neoplastic gastric tissue samples.RESULTS:Authors observed a significant reduction of 14-3-3εprotein expression in gastric cancer(GC)samples compared to their matched non-neoplastic tissue.Reduced levels of 14-3-3εwere also associated with diffuse-type GC and early-onset of this pathology.Our data suggest that reduced 14-3-3εmay have a role in gastric carcinogenesis process.CONCLUSION:Our results reveal that the reduced 14-3-3εexpression in GC and investigation of 14-3-3ε interaction partners may help to elucidate the carcino-genesis process.
