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国家自然科学基金(U0932001)

作品数:4 被引量:6H指数:1
相关作者:谢剑君吴健谊许丽艳蒋太峰杜则澎更多>>
相关机构:汕头大学南开大学更多>>
发文基金:国家自然科学基金中国博士后科学基金更多>>
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哺乳动物细胞CDK3系列表达载体的构建与表达被引量:1
2010年
目的:构建一系列细胞周期蛋白依赖激酶3(CDK3)在哺乳动物细胞的表达载体,并在真核细胞NIH3T3中进行初步表达。方法:从食管癌细胞中提取总RNA,RT-PCR扩增CDK3编码区,然后将PCR产物克隆到T载体;扩增后的CDK3片段分别亚克隆入pcDNA3-4、pNTAP和pEGFP等4种哺乳动物表达载体;将获得的表达载体转染小鼠成纤维细胞NIH3T3,并用蛋白免疫印迹法进行初步的CDK3表达分析。结果:PCR扩增获得约900 bp目的片段,经T载体克隆和DNA序列分析显示重组片段是人CDK3基因序列,全长915 bp;CDK3目的片段分别亚克隆入pcDNA3-4、pNTAP和pEGFP载体,获得相应表达载体;运用构建的表达载体,可以在真核细胞中表达出CDK3蛋白。结论:成功构建了CDK3系列哺乳动物细胞表达载体,并在NIH3T3中得到表达。
吴健谊蒋太峰许丽艳李恩民谢剑君
关键词:基因克隆真核表达载体
哺乳动物细胞CDK2系列表达载体的构建与表达被引量:1
2011年
目的:构建一系列细胞周期蛋白依赖激酶2(cyclin-dependent kinase 2,CDK2)在哺乳动物细胞的表达载体,为研究CDK2的功能和修饰提供实验材料。方法:从食管癌细胞中提取总RNA,逆转录PCR扩增CDK2编码区,然后将PCR产物克隆到T载体;扩增后的CDK2片段分别亚克隆入pcDNA3、pcDNA4、pNTAP和pEGFP等4种哺乳动物表达载体;最后,将获得的表达载体PEGFP/CDK2转染小鼠成纤维细胞NIH3T3进行初步的CDK2表达分析。结果:RT-PCR扩增获得约900 bp的目的片段,经T载体克隆和DNA序列分析,显示重组片段是人CDK2基因序列;CDK2片段分别亚克隆入上述4种载体后获得相应表达载体;运用构建的PEGFP/CDK2表达载体,在NIH3T3细胞中表达出CDK2蛋白。结论:成功构建了CDK2的哺乳动物细胞系列表达载体,并在NIH3T3细胞中成功表达目的蛋白。
吴健谊蒋太峰谢剑君张锴杜则澎许丽艳
关键词:CDK2基因克隆真核表达载体
Protein-coding genes combined with long noncoding RNA as a novel transcriptome molecular staging model to predict the survival of patients with esophageal squamous cell carcinoma被引量:4
2018年
Background:Esophageal squamous cell carcinoma(ESCC)is the predominant subtype of esophageal carcinoma in China.This study was to develop a staging model to predict outcomes of patients with ESCC.Methods:Using Cox regression analysis,principal component analysis(PCA),partitioning clustering,Kaplan-Meier analysis,receiver operating characteristic(ROC)curve analysis,and classification and regression tree(CART)analysis,we mined the Gene Expression Omnibus database to determine the expression profiles of genes in 179 patients with ESCC from GSE63624 and GSE63622 dataset.Results:Univariate cox regression analysis of the GSE63624 dataset revealed that 2404 protein-coding genes(PCGs)and 635 long non-coding RNAs(lncRNAs)were associated with the survival of patients with ESCC.PCA categorized these PCGs and lncRNAs into three principal components(PCs),which were used to cluster the patients into three groups.ROC analysis demonstrated that the predictive ability of PCG-lncRNA PCs when applied to new patients was better than that of the tumor-node-metastasis staging(area under ROC curve[AUC]:0.69 vs.0.65,P<0.05).Accord-ingly,we constructed a molecular disaggregated model comprising one lncRNA and two PCGs,which we desig-nated as the LSB staging model using CART analysis in the GSE63624 dataset.This LSB staging model classified the GSE63622 dataset of patients into three different groups,and its effectiveness was validated by analysis of another cohort of 105 patients.Conclusions:The LSB staging model has clinical significance for the prognosis prediction of patients with ESCC and may serve as a three-gene staging microarray.
Jin-Cheng GuoYang WuYang ChenFeng PanZhi-Yong WuJia-Sheng ZhangJian-Yi WuXiu-E XuJian-Mei ZhaoEn-Min LiYi ZhaoLi-Yan Xu
关键词:TRANSCRIPTOME
Fascin lysine 471 acetylation cooperates with serine 39 phosphorylation to inhibit actin-bundling activity and tumor metastasis in esophageal squamous cell carcinoma
2022年
Dear Editor,Fascin actin-bundling protein 1(Fascin,gene name:FSCN1)is the primary actin-bundling protein that clusters filaments into stable and bundled filopodia[1].In previous studies,elevated levels of Fascin promoted filopodia formation and enhanced the invasive ability of tumor cells,suggesting that Fascin is a target for therapeutic intervention[2,3].Therefore,it is necessary to elucidate the detailed molecular mechanism by which Fascin bundles F-actin filaments.
Fa-Min ZengYin-Wei ChengJian-Zhong HeXiu-E XuLian-Di LiaoLi-Yan XuEn-Min Li
关键词:FASCIN
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