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作 者:邓大炜 孔宪炳[1] 王平[1] 于庆三 王强[2]
机构地区:[1]重庆医科大学附属第一医院肝胆外科,重庆404100 [2]重庆医科大学附属第一医院胃肠外科,重庆404100
出 处:《中国生物化学与分子生物学报》2015年第4期422-428,共7页Chinese Journal of Biochemistry and Molecular Biology
基 金:重庆市科技攻关项目(No.9902200426-0001)~~
摘 要:探讨叉头框蛋白Q1(forkhead box Q1,FOXQ1)基因在肝癌中的临床意义及对肝癌细胞体外血管生成作用.利用qRT-PCR法及Western印迹法,检测24例肝癌、癌旁组织、正常肝细胞L02及肝癌细胞SMMC-7721中FOXQ1的mRNA和蛋白质的表达;利用免疫组织化学法检测68例肝癌及癌旁组织中FOXQ1的蛋白质表达.合成shRNA-FOXQ1及shRNA-NC慢病毒,转染到SMMC-7721细胞.用体外血管生成实验检测转染shRNA-FOXQ1的肝癌细胞血管生成能力.用qRT-PCR和Western印迹法检测细胞间FOXQ1、VEGF基因和蛋白质的表达.结果显示,癌组织和SMMC-7721细胞中FOXQ1 mRNA和蛋白质的表达均高于癌旁组织和正常肝细胞(P<0.05),FOXQ1蛋白的表达与TNM分期、肿瘤分化程度、肿瘤数目、肿瘤大小等参数差异显著(P<0.05).shRNA-FOXQ1组血管生成能力明显低于shRNA-NC组和空白组(P<0.05),FOXQ1、VEGF基因和蛋白质的表达也明显低于shRNA-NC组和空白组(P<0.05).研究结果证实,FOXQ1在肝癌中高表达,如果沉默FOXQ1的表达可抑制肝癌细胞血管生成,与肝癌的临床病理特征密切相关.To investigate the clinical significance of forkhead box Q1(FOXQ1) in human hepatocellular carcinoma(HCC),FOXQ1 mRNA and protein expression were detected by qRT-PCR and Western blotting in 24 pairs of HCC tissues,paracarcinoma tissues,hepatic cells(LO2) and SMMC-7721 cells.Immunohistochemistry was used for the detection of FOXQ1 protein levels in 68 pairs of HCC and corresponding paraneoplastic tissues.shRNA-FOXQ1 and shRNA-negative control(NC) were constructed and transfected into SMMC-7721 cells.The effect was detected by angiogenesis-related in vitro assay.The expression of FOXQ1 mRNA and protein in HCC were significantly higher than in paraneoplastic liver tissues.The expression of FOXQ1 in SMMC-7721 cells was also higher than that in LO2 liver cells(P〈0.05).The FOXQ1 protein level was correlated with TNM stage,tumor differentiation,tumor numbers and tumor size(P〈 0.05).FOXQ1 and VEGF expression were decreased following sh FOXQ1 transfection as compared with the shRNA-NC group(P〈0.05).These findingssuggested that FOXQ1 was upregulated in HCC.The abnormal expression of FOXQ1 related to the HCC progress and clinicopahology.Inhibition of FOXQ1 expression might suppress hepatoma cell angiopoiesis.
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