克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱。通过PCR扩增、Sanger测序等方法获得合作猪NTF3基因CDS区序列,使...克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱。通过PCR扩增、Sanger测序等方法获得合作猪NTF3基因CDS区序列,使用生物信息学方法分析其结构,并使用RT-qPCR技术检测其在不同组织中的表达水平。结果显示,合作猪NTF3基因CDS区长774 bp,编码257个氨基酸;等电点(pI)为9.46,属于碱性蛋白;合作猪与猪、牛亲缘关系较近;二级结构主要为α-螺旋和无规则卷曲,三级结构主要由β-折叠、α-螺旋等组成;整条链中亲水性氨基酸残基数量多于疏水性氨基酸残基数量,是亲水性蛋白;无跨膜区域,是非跨膜蛋白;在37~58位氨基酸残基有一个线圈结构域、64~79位有一个低复杂度结构域和144~249位有一个神经生长因子结构域;存在信号肽,推测是分泌蛋白;NTF3基因在合作猪脾脏的表达量最高,回肠表达量最低。展开更多
Objective Neurotrophin 3(NTF3)is involved in numerous biological processes;however,its role in hepatocellular carcinoma(HCC)is not well studied.This study investigated NTF3 function in HCC progression and revealed its...Objective Neurotrophin 3(NTF3)is involved in numerous biological processes;however,its role in hepatocellular carcinoma(HCC)is not well studied.This study investigated NTF3 function in HCC progression and revealed its underlying molecular mechanisms.Methods The prognostic relevance of NTF3 was determined through a bioinformatical analysis of publicly available TCGA data.Immunohistochemistry of HCC biopsies was performed to explore the expression of NTF3.Cell growth and proliferation were analyzed using a Cell Counting Kit-8(CCK-8)assay.Cell invasion and migration were analyzed using Boyden Transwell and wound healing assays.Protein expression and mRNA levels were evaluated through immunoblotting and quantitative polymerase chain reaction(qPCR).Cell apoptosis was evaluated with flow cytometry.Results NTF3 expression was significantly lower in HCC tissues than in adjacent non-tumor tissues.Low NTF3 expression was significantly associated with decreased patient survival and specific clinicopathological features.NTF3 overexpression reduced the proliferation,migration,and invasion abilities of HCC cell lines.Conclusion Decreased expression of NTF3 is associated with poor prognosis in HCC patients,likely due to its action in promoting HCC cell proliferation,migration,and invasion.Our findings provide a novel understanding into the pathogenesis of HCC and the role of NTF3 in tumor progression,suggesting that targeting NTF3 has potential therapeutic and diagnostic value for HCC.展开更多
文摘克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱。通过PCR扩增、Sanger测序等方法获得合作猪NTF3基因CDS区序列,使用生物信息学方法分析其结构,并使用RT-qPCR技术检测其在不同组织中的表达水平。结果显示,合作猪NTF3基因CDS区长774 bp,编码257个氨基酸;等电点(pI)为9.46,属于碱性蛋白;合作猪与猪、牛亲缘关系较近;二级结构主要为α-螺旋和无规则卷曲,三级结构主要由β-折叠、α-螺旋等组成;整条链中亲水性氨基酸残基数量多于疏水性氨基酸残基数量,是亲水性蛋白;无跨膜区域,是非跨膜蛋白;在37~58位氨基酸残基有一个线圈结构域、64~79位有一个低复杂度结构域和144~249位有一个神经生长因子结构域;存在信号肽,推测是分泌蛋白;NTF3基因在合作猪脾脏的表达量最高,回肠表达量最低。
基金Supported by grants from the National Natural Science Foundation of China(No.ZR2015HM025)Technology Development Program of Shandong Province,China(No.2011YD18003)CSCO-Hengrui Cancer Research Fund(No.Y-HR2018-293 and Y-HR2018-294).
文摘Objective Neurotrophin 3(NTF3)is involved in numerous biological processes;however,its role in hepatocellular carcinoma(HCC)is not well studied.This study investigated NTF3 function in HCC progression and revealed its underlying molecular mechanisms.Methods The prognostic relevance of NTF3 was determined through a bioinformatical analysis of publicly available TCGA data.Immunohistochemistry of HCC biopsies was performed to explore the expression of NTF3.Cell growth and proliferation were analyzed using a Cell Counting Kit-8(CCK-8)assay.Cell invasion and migration were analyzed using Boyden Transwell and wound healing assays.Protein expression and mRNA levels were evaluated through immunoblotting and quantitative polymerase chain reaction(qPCR).Cell apoptosis was evaluated with flow cytometry.Results NTF3 expression was significantly lower in HCC tissues than in adjacent non-tumor tissues.Low NTF3 expression was significantly associated with decreased patient survival and specific clinicopathological features.NTF3 overexpression reduced the proliferation,migration,and invasion abilities of HCC cell lines.Conclusion Decreased expression of NTF3 is associated with poor prognosis in HCC patients,likely due to its action in promoting HCC cell proliferation,migration,and invasion.Our findings provide a novel understanding into the pathogenesis of HCC and the role of NTF3 in tumor progression,suggesting that targeting NTF3 has potential therapeutic and diagnostic value for HCC.
