解剖学报

解剖学报 ›› 2021, Vol. 52 ›› Issue (6): 909-912.doi: 10.16098/j.issn.0529-1356.2021.06.010

• 肝再生与细胞周期调控 • 上一篇    下一篇

大鼠肝再生终止阶段肝细胞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 高寒1,2 郭建林 1,2 靳伟1,2 常翠芳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:2021-01-04 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: 徐存拴 E-mail:cellkeylab@126.com

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 termination

ZANG Xia-yan1, 2 WANG Zi-hui1, 2 GAO Han1, 2 GUO Jian-lin1, 2 JIN Wei1,2 CHANG Cui-fang1, 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:2021-01-04 Revised:2021-03-12 Online:2021-12-06 Published:2021-12-06
  • Contact: XU Cun-shuan E-mail:cellkeylab@126.com

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摘要：

目的 了解大鼠肝再生终止阶段CCAAT增强子结合蛋白α（CEBPα）mRNA、miR-144-3p、rno-LOC100365958_0001、rno-Usp3_0010和rno-AC241873_0010调节肝细胞进入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6 h和72 h时，CEBPα mRNA的比值为1.12±0.27和2.52±0.15，miR-144-3p为2.97±0.81和0.99±0.36，rno-LOC100365958_0001为0.16±0.05和6.78±0.62，rno-Usp3_0010为0.13±0.01和7.61±0.69，rno-AC241873_0010为0.09±0.02和7.53±0.69。CEBPα促进的G1期相关基因细胞周期蛋白G2（CCNG2）为0.25±0.06和1.10±0.22，抑制的G1期相关基因ETS变异转录因子6（ETV6）为2.22±0.68和0.99±0.27，MAPK14为2.01±0.32和0.85±0.11，信号转导子和转录激活因子3（STAT3）为2.88±0.24和1.08±0.07，肿瘤坏死因子（TNF）为2.29±0.51和0.86±0.51。CEBPα促进的G0期相关基因G0/G2期开关2（G0S2）为0.64±0.08和4.71±0.91，磷脂酶A2-IVA（PLA2G4A）为0.42±0.13和2.83±0.62。   结论  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。相反，PH后72 h时，CEBPα mRNA上调，有利于CEBPα促进G0期相关基因表达和肝细胞处于G0期。

关键词: 部分肝切除, 肝再生, G0期肝细胞, G1期肝细胞, CCAAT增强子结合蛋白α, 生物高通量检测, 竞争性内源RNA综合分析, 大鼠

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 to enter 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 et al, the large-scale quantitative detection of ceRNAs was processed by the high-throughput biotechnology, the interaction network of competitive endogenous RNAs(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 6 hours and 72 hours after PH, the ratio value of CEBPα mRNA shows 1.12±0.27 and 2.52±0.15, miR-144-3p displays 2.97±0.81 and 0.99±0.36, rno-LOC100365958_0001 indicates 0.16±0.05 and 6.78±0.62, rno-Usp3_0010 shows 0.13±0.01 and 7.61±0.69, rno-AC241873_0010 indicates 0.09±0.02 and 7.53±0.69. The G1 phase-related gene cylin G2(CCNG2) promoted by CEBPα 0.25±0.06 and 1.10±0.22, but the G1 phase-related genes inbibited by CEBPα were following, ETS variant transcription factor 6(ETV6) 2.22±0.68 and 0.99±0.27, MAPK14 2.01±0.32 and 0.85±0.11, the signal transducer and activator of transcription 3(STAT3)为2.88±0.24 and 1.08±0.07, tumor necrosis factor(TNF) 2.29±0.51 and 0.86±0.51. The G0 phase-related genes promoted by CEBPα were following, G0/G1 witch 2(G0S2) 0.64±0.08 and 4.71±0.91, phospholipase A2 group IVA(PLA2G4A) 0.42±0.13 and 2.83±0.62.   Conclusion  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. On the contrary, CEBPα mRNA is up-regulated at 72 hours 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.

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

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