| Liver regeneration（LR）of rats after 2/3 hepatectomy（PH）is a complex and orderly process,which includes the initiation（0～6 h after PH）,proliferation（6～72 h after PH）and termination（72～168 h after PH）.Former studies have shown that competitive endogenousRNA（ceRNA）plays a crucial role in rat LR.At present,liver resection and liver transplantation in clinical liver diseases treatmentmainly rely on the regeneration ability of liver,whereas,the activation and completion of the liver’s potent regenerative capacity is closely related to the G0/G1 phase transition and the following proliferation of hepatocytes.Therefore,a study on liver regeneration,especially the mechanism that ceRNA promotes or inhibits the G0/G1 phase transition for regulating liver cell proliferation,can shed light on the clinical treatment of liver diseases.In view of this,we conducted a research on the interactions amongst circRNA,miRNA and mRNA,aiming to elucidate the merchanism underlying the G0/G1 regulation of hepatocytes.In the study,we constructed 2/3 hepatectomy model,then use bioinformatic and high-throughput detection technology to analyze the expression of ceRNA during rat LR.Afterwards,we retrieved G0/G1 phase-related mRNAs through NCBI database alignments,innovative pathway analysis（IPA）,and constructed a candidate mRNA set.We got miRNAs relating to these mRNAs with alignment of mi Rwalk database.Similarly,we got circRNAs relating to the miRNAs through mi Randa and Target Scan database search,and further screen out the G0/G1-related circRNA.With integrating the above-mentioned G0/G1-related ceRNA and the high-throughput transcriptomic sequencing results,we obtained G0/G1-related ceRNA with available expression data.We then carried out a series of analysis as below: Using NCBI and Animal TFDB database alignments to screen out transcription factors;define differential expressedRNA（DERNA）via|log2Foldchange|≥1 and p≤0.05;select crucial gene Cebpz via GSEA software taking TOP p-value as condition.Then,we selected the upstream miRNA and circRNA of Cebpz according to the above results.Next,we used the Cistrome Data Browser database search and IPA software to get the downstream genes regulated by Cebpz,and finally took advantage of Cytoscape 3.2 software to construct the circRNA-miRNA-Cebpz interaction network.This study revealed that there were 13,251 types of mRNA,921 types of miRNA,and13,999 types of circRNA were detected.Among them,differentially expressed circRNA,miRNA,and mRNA in rat liver regeneration are 2843,348,and 2479 types,respectively;there are 552,349,and 745 related circRNA,miRNA,and mRNA in the G0 phase and G1 phase,respectively.Related circRNA,miRNA and mRNA are 229,104 and 177 kinds,respectively.Cytoscape 3.2software constructed and analyzed the Cebpz-related ceRNA interaction network and found that Cebpz formed 464 interactions with 30 miRNAs and 123 circRNAs in G0 phase,and 720 interactions with 43 miRNAs and 189 circRNAs in G1 phase.After selecting the crucial mi R-136-3p via ceRNA integrated analysis method,combining expression,function and early research results of mi R-136-3p,four circRNAs showed crucial roles in the G0 and G1 phases of rat LR,they are: circ02548,circ02174,circ10034,and circ16474.By expression variation,we got five downstream genes regulated by CEBPz: Tom1,Vcp,Cdk2ap2,Creb3l4,and Txnip.The study disclosed that Cebpz may regulate downstream gene expression through circ02548-mi R-136-3p-Cebpz,circ02174-mi R-136-3p-Cebpz,circ10034-mi R-136-3p-Cebpz and circ16474-mi R-136-3p-Cebpz interaction,and further determine the hepatocytes in G0 or in G1 phase.This paper studies the expression changes and effects of circRNA,miRNA and mRNA in the G0 and G1 phases of rat LR,and analyzes the molecular mechanism that they regulate hepatocytes in G0 phase or G1 phase through the ceRNA network,in order to further reveal the rat liver the regulation mechanism of regenerative cell cycle,cell proliferation and the engineering of new target drugs provide the basis and ideas. |
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