大鼠肝再生启动阶段肝细胞CCAAT增强子结合蛋白α mRNA、微小RNA-144-3p和3种环状RNA的表达和作用

徐存拴

doi:10.16098/j.issn.0529-1356.2021.06.009

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解剖学报 ›› 2021, Vol. 52 ›› Issue (6) : 901-908. DOI: 10.16098/j.issn.0529-1356.2021.06.009

肝再生与细胞周期调控

大鼠肝再生启动阶段肝细胞CCAAT增强子结合蛋白α mRNA、微小RNA-144-3p和3种环状RNA的表达和作用

臧夏炎^1,2 王子慧^1,2李亚霏^1,2 郭建林^1,2 靳伟^1,2常翠芳^1,2徐存拴^1,2*

作者信息 +

Expression and role of CCAAT enhancer binding protein α mRNA, microRNA-144-3p and three kinds of circular RNAs of hepatocytes during the rat liver regeneration initiation

ZANG Xia-yan^{1, 2} WANG Zi-hui^{1, 2} LI Ya-fei^{1, 2} GUO Jian-lin^{1, 2} JIN Wei^{1, 2}CHANG Cui-fang³ XU Cun-shuan^{1, 2*}

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文章历史 +

摘要

目的了解大鼠肝再生启动阶段CCAAT增强子结合蛋白α（CEBPα） mRNA、微小RNA（miR）-144-3p、rno-LOC100365958_0001、rnoUsp3_0010和rno-AC241873_0010调节肝细胞处于G0期还是G1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竞争性内源RNA（ceRNA）表达的变化，用Cytoscape 3.2软件构建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， CEBPα mRNA的比值为4.88±0.78和1.12±0.27，miR-144-3p为1.30±0.11和2.97±0.81，rno-LOC100365958_0001为6.99±0.55和0.16±0.05，rno-Usp3_0010为8.48±0.67和0.13±0.01，rno-AC241873_0010为11.85±0.93和0.09±0.02。CEBPα促进的G0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，G0/G1开关2（G0S2）为3.85±0.23和0.64±0.08，信号转导子和转录激活因子1（STAT1）为2.57±0.13和1.32±0.13，转化生长因子β受体2（TGFBR2）为2.77±0.20和0.79±0.13，抑制的G0期相关基因细胞周期蛋白依赖性激酸酶抑制剂1a（CDKN1a）为0.39±0.07和0.93±0.15。CEBPα促进的G1期相关基因细胞周期蛋白G2（CCNG2）为1.19±0.09和0.25±0.06，抑制的G1期相关基因ETS变异转录因子6（ETV6）为0.77±0.05和2.22±0.68，血红蛋白加氧酶1（HMOX1）为1.05±0.21和4.57±0.88，MAPK14为1.01±0.15和2.01±0.32，STAT3为0.74±0.15和2.88±0.24，肿瘤坏死因子（TNF）为0.80±0.14和2.29±0.51。结论 PH后0 h时，CEBPα mRNA上调，有利于CEBPα促进的G0期相关基因表达和肝细胞处于G0期。相反，PH后6 h时，rno-LOC100365958_0001、rno-Usp3_0010、rno-AC241873_0010和miR-144-3p的相互作用加强了后者对CEBPα mRNA的抑制，不利于CEBPα形成，有利于CEBPα抑制的G1期相关基因表达和肝细胞处于G1期。

Abstract

Objective To explore the pathways and patterns which CCAAT enhancer binding proteinα(CEBPα) mRNA, miR-144-3p, rno-LOC100365958_0001, rno-Usp3_0010 and rno-AC241873_0010 regulate the hepatocytes in G0 phase and G1 phase during rat liver regeneration (LR). Methods The rat 2/3 hepatectomy (partial hepatectomy, PH) model was prepared as described by Higgins, the hepatocytes of rat liver right lobes were isolated in 9∶00-11∶00 am according to the method of Smedsrod, the large-scale quantitative detection of competitive endogenous RNAs(ceRNAs) was processed by the high-throughput biotechnology, the interaction network of ceRNAs was constructed by Cytoscape 3.2 software, and the correlation in expression and role of ceRNAs was analyzed by ceRNA comprehensive analysis. Results It was found that at 0 hour and 6 hours after PH, the ratio value of CEBPα mRNA showed 4.88±0.78 and 1.12±0.27, miR-144-3p displays 1.30±0.11 and 2.97±0.81, rno-LOC100365958_0001 indicateed 6.99±0.55 and 0.16±0.05, rno-Usp3_0010 shows 8.48±0.67 and 0.13±0.01, rno-AC241873_0010 indicates 11.85±0.93 and 0.09±0.02. The G0 phase-related genes promoted by CEBPα were following, cyclin dependent kinase 2 associated protein 2(CDK2AP2) 2.55±0.42 and 0.74±0.11, G0/G1 swich 2(G0S2) 3.85±0.23 and 0.64±0.08, the signal transducer and activator of transcription 1(STAT1) 2.57±0.13 and 1.32±0.13, transforming growth factor beta receptor 2(TGFBR2) 2.77±0.20 and 0.79±0.13, but the G0 phase-related gene cyclin dependent kinase inhibitor 1a(CDKN1a) inbibited by CEBPα 0.39±0.07 and 0.93±0.15. The G1 phase-related gene cyclin G2(CCNG2) promoted by CEBPα 1.19±0.09 and 0.25±0.06, but the G1 phase-related genes inbibited by CEBPα were following, ETS variant transcription factor 6(ETV6) 0.77±0.05 and 2.22±.68, hemoglobin oygenase 1(HMOX1) 1.05±0.21 and 4.57±0.88, MAPK14 1.01±0.15 and 2.01±0.32, STAT3 0.74±0.15 and 2.88±0.24, tumor necrosis factor(TNF) 0.80±0.14 and 2.29±0.51. Conclusion CEBPα mRNA is up-regulated at 0 h after PH, that is helpful for the expression of the G0 phase-related genes promoted by CEBPα and for the hepatocytes to be in G0 phase. On the contrary, the interaction of miR-144-3p and rno-LOC100365958_0001, rno-Usp3_0010 and rno-AC241873_0010 leads to CEBPα mRNA to bind with miR-144-3p, to CEBPα being formed difficultly, and to the expression of the G1 phase-related genes inbibited by CEBPα probablely, and to the hepatocytes being in G1 phase at 6 hours after PH.

导出引用

臧夏炎王子慧李亚霏郭建林靳伟常翠芳徐存拴. 大鼠肝再生启动阶段肝细胞CCAAT增强子结合蛋白α mRNA、微小RNA-144-3p和3种环状RNA的表达和作用[J]. 解剖学报. 2021, 52(6): 901-908 https://doi.org/10.16098/j.issn.0529-1356.2021.06.009

ZANG Xia-yan WANG Zi-hui LI Ya-fei GUO Jian-lin JIN WeiCHANG Cui-fang XU Cun-shuan. Expression and role of CCAAT enhancer binding protein α mRNA, microRNA-144-3p and three kinds of circular RNAs of hepatocytes during the rat liver regeneration initiation[J]. Acta Anatomica Sinica. 2021, 52(6): 901-908 https://doi.org/10.16098/j.issn.0529-1356.2021.06.009

中图分类号： Q257

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