Hsa_circ_0009096/miR-370-3p modulates hepatic stellate cell proliferation and fibrosis during biliary atresia...

Area of Science:

• Hepatology
• Molecular Biology
• Cell Biology

Background:

• Hepatic stellate cell (HSC) activation and fibrosis are central to biliary atresia (BA) pathogenesis.
• The molecular mechanisms underlying HSC activation and fibrosis in BA remain poorly understood.

Purpose of the Study:

• To investigate the role of circRNA hsa_circ_0009096 in regulating HSC proliferation and hepatic fibrosis.
• To elucidate the molecular mechanism by which hsa_circ_0009096 influences BA.

Main Methods:

• Established a cellular hepatic fibrosis model using LX-2 cells treated with transforming growth factor β (TGF-β1).
• Assessed hsa_circ_0009096 stability using RNaseR and actinomycin D assays.
• Quantified expression of hsa_circ_0009096, miR-370-3p, and target genes via reverse transcription-qPCR.
• Validated direct binding of hsa_circ_0009096 to miR-370-3p using dual luciferase reporter assay.
• Analyzed cell cycle progression, apoptosis, and protein levels of α-SMA, COL1A1, and TGFBR2 using flow cytometry, immunocytochemistry, and western blotting.

Main Results:

• Hsa_circ_0009096 demonstrated significant stability and its expression increased upon TGF-β1 treatment in LX-2 cells.
• Hsa_circ_0009096 directly binds to miR-370-3p, suppressing its activity and promoting TGFBR2 expression.
• Knockdown of hsa_circ_0009096 inhibited HSC proliferation, promoted apoptosis, and reduced fibrotic markers (α-SMA, COL1A1) in TGF-β1-treated cells.
• Inhibition of miR-370-3p reversed the effects of hsa_circ_0009096 knockdown on cell cycle, apoptosis, and fibrotic markers.

Conclusions:

• Hsa_circ_0009096 promotes HSC proliferation and hepatic fibrosis in BA pathogenesis.
• This promotion is mediated by the sponging of miR-370-3p, leading to accelerated TGFBR2 expression.
• Hsa_circ_0009096 represents a potential therapeutic target for biliary atresia.