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Updated: Feb 2, 2026
Seven Steps to Stellate Cells
Published on: May 10, 2011
Thymosin-β4 Mediates Hepatic Stellate Cell Activation by Interfering with CircRNA-0067835/miR-155/FoxO3 Signaling
Lili Zhu1,2, Tingting Ren1, Zixin Zhu3
1Department of Infectious Diseases, The Affiliated Hospital of Guizhou Medical University, Guizhou, China.
Background/Aims:
Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrosis. Our study proved that thymosin beta 4 (Tβ4) has anti-fibrogenic effects in HSCs through PI3K/AKT pathway. However, the underlying mechanisms are not fully elucidated. Circular RNAs (circRNAs) play important roles in fine-tuning gene expression and are often deregulated in cancers. However, the expression profile and clinical significance of in liver fibrosis is still unknown. Therefore, we hypothesize that Tβ4 influences circRNAs in liver fibrosis.
Methods:
Circular RNA microarray was conducted to identify Tβ4-related circRNAs. Pathway analysis and miRNA response elements analysis was conducted to predict the potential roles of differentially expressed circRNAs in liver fibrosis. CCK8 assays and flow cytometric assays were conducted to clarify the role of circRNA in liver fibrosis. Bioinformatics analysis and in vitro experiments were conducted to clarify the mechanism of circRNA-mediated gene regulation in liver fibrosis.
Results:
A total of 644 differentially expressed circRNAs were identified between the Tβ4-depleted LX-2 cells and the control LX2 cells. The expression of circRNA-0067835 was significantly increased in the Tβ4-depleted LX-2 cells compared with control. Knockdown of circRNA-0067835 observably decreased LX-2 cell proliferation by causing G1 arrest and promoting apoptosis. Bioinformatics online programs predicted that circRNA-0067835 acted as miR-155 sponge to regulate FOXO3a expression, which was validated using luciferase reporter assay.
Conclusion:
Our experiments showed that circRNA-0067835 regulated liver fibrosis progression by acting as a sponge of miR-155 to promote FOXO3a expression, indicating that circRNA-0067835 may serve as a potential therapeutic target for patients with liver fibrosis.
Insights
Thymosin beta 4 (Tβ4) impacts liver fibrosis by regulating circRNA-0067835. This circular RNA acts as a sponge for miR-155, promoting FOXO3a expression and influencing fibrosis progression.
Area of Science:
- Hepatology and molecular biology
- Cellular and molecular mechanisms of liver fibrosis
- Circular RNA biology
Background:
- Hepatic stellate cells (HSCs) are key drivers of liver fibrosis.
- Thymosin beta 4 (Tβ4) exhibits anti-fibrogenic effects in HSCs via the PI3K/AKT pathway.
- The precise mechanisms by which Tβ4 influences liver fibrosis, particularly involving circular RNAs (circRNAs), remain incompletely understood.
Purpose of the Study:
- To investigate the role of circRNAs in Tβ4-mediated regulation of liver fibrosis.
- To identify specific circRNAs affected by Tβ4 depletion in HSCs.
- To elucidate the molecular mechanisms underlying circRNA involvement in liver fibrosis progression.
Main Methods:
- Circular RNA microarray analysis to identify differentially expressed circRNAs upon Tβ4 depletion.
- Bioinformatics and pathway analysis to predict circRNA functions.
- In vitro experiments including CCK8 assays, flow cytometry, and luciferase reporter assays to validate circRNA roles and mechanisms.
Main Results:
- A total of 644 differentially expressed circRNAs were identified between Tβ4-depleted and control LX-2 cells.
- CircRNA-0067835 expression was significantly upregulated in Tβ4-depleted cells.
- Knockdown of circRNA-0067835 inhibited LX-2 cell proliferation, induced G1 arrest, and promoted apoptosis.
- CircRNA-0067835 was validated as a miR-155 sponge that regulates FOXO3a expression.
Conclusions:
- CircRNA-0067835 plays a critical role in regulating liver fibrosis progression.
- It functions by sequestering miR-155, thereby increasing FOXO3a expression.
- CircRNA-0067835 represents a potential therapeutic target for liver fibrosis.
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