Mechanistic study on circVAPA promoting the Hippo signaling pathway to inhibit liver regeneration via miR-101a-3p/TEAD3 axis
10.3969/j.issn.1671-8348.2025.05.002
- VernacularTitle:circVAPA通过miR-101a-3p/TEAD3轴促进Hippo通路抑制肝再生的机制研究
- Author:
Jian ZHAO
1
;
Yunhong DAI
;
Yanli DANG
Author Information
1. 云南大学附属医院普外二科,昆明 650021
- Keywords:
circular RNA;
circVAPA;
miR-101a-3p/TEAD3 axis;
Hippo signaling pathway;
liver regen-eration
- From:
Chongqing Medicine
2025;54(5):1050-1058
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the molecular mechanism of circular RNA(circ)VAPA promoting the Hippo pathway to inhibit liver regeneration through the miR-101a-3p/TEAD3 axis.Methods A mouse model of 70％partial hepatectomy-induced liver regeneration was constructed,and the expressions of circVA-PA,miR-101a-3p,and TEAD3 were analyzed.Mouse embryonic hepatocyte BNL CL.2 cells were transfected with siRNA or overexpression plasmids and divided into the VAPA-NC group,the VAPA-NC+miR-101a-3p mimic group,the VAPA-NC+miR-101a-3p mimic-NC group,the VAPA-OE group,the VAPA-OE+miR-101a-3p mimic group and the VAPA-OE+miR-101a-3p mimic-NC group.CCK-8 method and flow cytometry were used to analyze the proliferation,apoptosis,and cell cycle changes of hepatocytes.Immunofluorescence staining was used to analyze the nuclear translocation of YAP1.Quantitative Reverse Transcription Real-Time PCR(qRT-PCR)was used to analyze the expressions of key genes in the Hippo pathway.The dual-lu-ciferase reporter gene assay was used to verify the targeting relationships between circVAPA and miR-101a-3p,as well as between miR-101a-3p and TEAD3.Results The level of circVAPA gradually increased during liver regeneration(P＜0.05),while the level of miR-101a-3p first increased and then decreased(P＜0.05).When miR-101a-3p was overexpressed alone,the cell proliferation rate was the highest(P＜0.05),but it had no effect on the cell apoptosis rate(P＞0.05).When circVAPA was overexpressed alone,it had no effect both on the cell proliferation rate and apoptosis(P＞0.05).After co-overexpression of circVAPA and miR-101a-3p,the cell proliferation rate significantly decreased,and the cell apoptosis significantly increased(P＜0.05).When miR-101a-3p was overexpressed alone,a large number of cells entered into the S phase.After co-overex-pression of circVAPA and miR-101a-3p,a large number of cells were blocked in the G2/M phase.The phos-phorylation level of the Hippo upstream gene YAP1 significantly increased at 6 hours after liver regeneration(P＜0.05)and then rapidly decreased.However,co-overexpression of circVAPA and miR-101a-3p did not af-fect the level of p-YAP1 and the nuclear translocation of YAP1(P＞0.05).The expression levels of the Hip-po downstream gene CTGF and the transcription factor TEAD3 first increased and then decreased during liver regeneration(P＜0.05),and there was no significant change in CYR61(P＞0.05).After co-overexpression of circVAPA and miR-101a-3p,the expression level of CTGF increased(P＜0.05).Knocking down or overex-pressing circVAPA did not affect the expression of TEAD3(P＞0.05),while overexpressing miR-101a-3p could significantly inhibit the expression level of TEAD3(P＜0.05).The dual-luciferase reporter gene assay confirmed the targeting relationships between circVAPA and miR-101a-3p,as well as between miR-101a-3p and TEAD3.Conclusion circVAPA promotes the Hippo pathway to inhibit liver regeneration through the miR-101a-3p/TEAD3 axis.
