解剖学报

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 • 上一篇    下一篇

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*   

  1. 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
  • 收稿日期:2021-01-11 修回日期:2021-03-12 出版日期:2021-12-06 发布日期:2021-12-06
  • 通讯作者: 郭建林;徐存拴 E-mail:cellkeylab@126.com

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 Han1,2 WANG Zi-hui1,2 ZANG Xia-yan1,2 JIN Wei1,2  CHANG Cui-fang1,2 GUO Jian-lin1,2* XU Cun-shuan1,2*   

  1. 1. College of Life Science, He’nan Normal University, He’nan Xinxiang 453007, China; 2. State Key Laboratory Cultivation Base for Cell Differentiation Regulation, He’nan Xinxiang 453007, China
  • Received:2021-01-11 Revised:2021-03-12 Online:2021-12-06 Published:2021-12-06
  • Contact: GUO Jian-lin;XU Cun-shuan E-mail:cellkeylab@126.com

PDF

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。

关键词: 肝再生, G0期肝细胞, G1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

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.

Key words: Liver regeneration, Hepatocyte in G0 phase Hepatocyte in G1 phase,  , CCAAT enhancer binding protein ζ, Competitive endogenous RNA comprehensive aalysis, Biological high-throughput detection, Competitive endogenous RNA comprehensive analysis, Rat

中图分类号: