大鼠肝再生启动阶段肝细胞CCAAT增强子结合蛋白ζ mRNA、微小RNA-136-3p和4种环状RNA的表达和作用  

Expression and role of CCAAT enhancer binding protein ζ mRNA,microRNA-136-3p and four kinds of circular RNAs of hepatocytes during the rat liver regeneration initiation

作　　者：高寒 王子慧 臧夏炎 靳伟[1,2] 常翠芳[1,2] 郭建林[1,2] 徐存拴 GAO Han;WANG Zi-hui;ZANG Xia-yan;JIN Wei;CHANG Cui-fang;GUO Jian-lin;XU Cun-shuan(College of Life Science,He’nan Normal University,He'nan Xinxiang 453007,China;State Key Laboratory Cultivation Base for Cell Differentiation Regulation,He'nan Xinxiang 453007,China)

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]河南省-科技部共建细胞分化调控国家重点实验室培育基地,河南新乡453007

出　　处：《解剖学报》2021年第6期921-924,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金(31572270,81771226);国家青年自然科学基金(U1804186,U1404312,31601038,31201093,81200317)。

摘　　要：目的了解大鼠肝再生启动阶段CCAAT增强子结合蛋白ζ(CEBPζ)mRNA、miR-136-3p、rno-Got1_0001、rno-Crebrf_0009、rno-Slc38a9_0001和rno-LOC100362999_0001调节肝细胞处于G_(0)期还是G_(1)期的途径和方法。方法按Higgins等方法制备大鼠2/3肝切除(PH)模型,按Smedsrod等方法分离肝细胞,用大规模定量分析技术检测大鼠肝再生中肝细胞的ceRNA表达变化,用Cytoscape 3.2软件构建竞争性内源RNA(ceRNA)的相互作用网络,用ceRNA综合分析等方法解析它们表达的相关性和作用相关性。结果PH后0 h和6 h时,CEBPζmRNA的比值为0.97±0.15和2.56±0.12,miR-136-3p为1.05±0.32和0.38±0.04,rno-Got1_0001为0.33±0.03和4.35±0.78,rnoCrebrf_0009为1.17±0.32和2.99±0.28,rno-Slc38a9_0001为0.67±0.08和2.64±0.29,rno-LOC100362999_0001为0.25±0.02和0.92±0.22。CEBPζ抑制的G_(0)期相关基因细胞周期蛋白依赖性激酶2相关蛋白2(CDK2AP2)为2.55±0.42和0.74±0.11,myb1膜运输蛋白的靶标(TOM1)为2.80±0.91和1.40±0.36,含缬草肽蛋白质(VCP)为2.63±0.17和1.10±0.10。CEBPζ促进的G_(1)期相关基因c AMP响应元件结合蛋白3样4(CREB3L4)为1.13±0.63和2.00±0.81,硫氧还蛋白相互作用蛋白(TXNIP)为1.03±0.07和2.50±0.19。结论PH后0 h时,CEBPζmRNA未上调,有利于CEBPζ抑制G_(0)期相关基因的表达和肝细胞处于G_(0)期。相反,PH后6 h时,rno-Got1_0001、rno-Crebrf_0009、rnoSlc38a9_0001、rno-LOC100362999_0001和miR-136-3p的相互作用解除了后者对CEBPζmRNA的抑制,有利于CEBPζ的形成,有利于CEBPζ促进G_(1)期相关基因的表达和肝细胞处于G_(1)期。Objective To explore the pathways and patterns which the CCAAT enhancer binding proteinζ(CEBPζ)mRNA,miR-136-3p,rno-Crebrf_0009,rno-Slc38a9_0001,rno-LOC100362999_0001 and rno-Got1_0001regulate the hepatocytes in G_(0) phase and G_(1) phase during rat liver regeneration(LR).Methods The rat 2/3 partial hepatectomy(PH)model was prepared as described by Higgins,the hepatocytes of rat liver right lobes were isolated in9∶00-11∶00 am according to the method of Smedsrod et al,the large-scale quantitative detection of competitive endogenous RNA(ceRNAs)was processed by the high-throughput biotechnology,the interaction network of ceRNA was constructed by Cytoscape 3.2 software,and the correlation in expression and role of ceRNA was analyzed by ceRNA comprehensive analysis.Results It was found that at 0 hour and 6 hours after PH,the ratio value of CEBPζmRNA shows 0.97±0.15and 2.56±0.12,miR-136-3p displays 1.05±0.32 and 0.38±0.04,rno-Got1_0001 indicates 0.33±0.03 and 4.35±0.78,rno-Crebrf_0009 shows 1.17±0.32 and 2.99±0.28,rno-Slc38a9_0001 indicates 0.67±0.08 and 2.64±0.29,rnoLOC100362999_0001 shows 0.25±0.02 and 0.92±0.22.The G_(0)phase-related genes inhibited by CEBPζwere following,the cyclin dependent kinase 2 associated protein 2(CDK2AP2)2.55±0.42 and 0.74±0.11,the target of myb1 membrane trafficking protein(TOM1)2.80±0.91 and 1.40±0.36,the valosin containing protein(VCP)2.63±0.17 and 1.10±0.10.The G_(1)phase-related genes prometed by CEBPζwere following,the c AMP responsive element binding protein 3like 4(CREB3L4)1.13±0.63 and 2.00±0.81,the thioredoxin interacting protein(TXNIP)1.03±0.07 and 2.50±0.19.Conclusion CEBPζmRNA is not up-regulated at 0 hour after PH,that is helpful for the expression of the G_(0)phaserelated genes inhibited by CEBPζand for the hepatocytes to be in G_(0)phase.On the contrary,the interaction of miR-136-3p and rno-Got1_0001,rno-Crebrf_0009,rno-Slc38a9_0001 and rno-LOC100362999_0001 leads to CEBPζmRNA to bind with miR-136-3p,to CEBPζbeing formed probablely,and to t

关 键 词：肝再生 G_(0)期肝细胞 G_(1)期肝细胞 CCAAT增强子结合蛋白ζ 竞争性内源RNA综合分析 生物高通量检测 大鼠 

分 类 号：Q257[生物学—细胞生物学]