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

徐存拴

doi:10.16098/j.issn.0529-1356.2021.06.013

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解剖学报 ›› 2021, Vol. 52 ›› Issue (6) : 921-624. DOI: 10.16098/j.issn.0529-1356.2021.06.013

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ζ mRNA、微小RNA-136-3p和4种环状RNA的表达和作用

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

作者信息 +

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

GAO Han^1,2 WANG Zi-hui^1,2 ZANG Xia-yan^1,2 JIN Wei^1,2 CHANG Cui-fang^1,2 GUO Jian-lin^1,2* 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ζ（CEBPζ）mRNA、miR136-3p、rno-Got1_0001、rno-Crebrf_0009、rno-Slc38a9_0001和rno-LOC100362999_0001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，CEBPζ mRNA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，rno-Crebrf_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，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G1期相关基因cAMP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，CEBPζ mRNA未上调，有利于CEBPζ抑制G0期相关基因的表达和肝细胞处于G0期。相反，PH后6 h 时，rno-Got1_0001、rno-Crebrf_0009、rno-Slc38a9_0001、rno-LOC100362999_0001和miR-136-3p的相互作用解除了后者对CEBPζ mRNA的抑制，有利于CEBPζ的形成，有利于CEBPζ促进G1期相关基因的表达和肝细胞处于G1期。

Abstract

Objective To explore the pathways and patterns which the CCAAT enhancer binding protein ζ (CEBPζ) mRNA, miR-136-3p, rno-Crebrf_0009, rno-Slc38a9_0001, rno-LOC100362999_0001 and rno-Got1_0001 regulate the hepatocytes in G0 phase and G1 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 in 9∶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 CEBPζ mRNA shows 0.97±0.15 and 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, rno-LOC100362999_0001 shows 0.25±0.02 and 0.92±0.22. The G0 phase-related genes inhibited by CEBPζ were 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 G1 phase-related genes prometed by CEBPζ were following, the cAMP responsive element binding protein 3 like 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 CEBPζ mRNA is not up-regulated at 0 hour after PH, that is helpful for the expression of the G0 phase-related genes inhibited by CEBPζ and for the hepatocytes to be in G0 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 CEBPζ mRNA to bind with miR-136-3p, to CEBPζ being formed probablely, and to the expression of the G1 phase-related genes promoted by CEBPζ probablely, and to the hepatocytes being in G1 phase at 6 hours after PH.

导出引用

高寒王子慧臧夏炎靳伟常翠芳郭建林徐存拴. 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[J]. 解剖学报. 2021, 52(6): 921-624 https://doi.org/10.16098/j.issn.0529-1356.2021.06.013

GAO Han WANG Zi-hui ZANG Xia-yan JIN Wei CHANG Cui-fang GUO Jian-lin XU Cun-shuan. 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[J]. Acta Anatomica Sinica. 2021, 52(6): 921-624 https://doi.org/10.16098/j.issn.0529-1356.2021.06.013

中图分类号： Q257

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