Y-box binding protein-1 modulates hepatic progenitor cells proliferation via Gli2

Abstract

Abstract: Objective To explore the effect of Y-box binding protein-1 (YB-1) on the proliferation of hepatic progenitor cells (HPCs) and the molecular mechanism.Methods The lentiviral vector system was adopted to down-regulate YB-1 expression. Cell counting kit 8 (CCK-8) and flow cytometry were performed to assess the proliferation rate. RNA-sequencing was employed to evaluate the effect of YB-1 on gene expression of HPCs. Chromatin immunoprecipitation coupled with high-throughput deep sequencing (ChIP-Sequence) was performed to detect the potential target genes of YB-1 in HPCs. ChIP-PCR was conducted to identify the binding of YB-1 and Gli2. Luciferase reporter assay was conducted to analyze the regulatory effect of YB-1 on Gli2 transcription. Gli1/Gli2 inhibitor GANT61 was co-culture with HPCS to assess the effect of Gli2 on HPCs proliferation. Results When YB-1 mRNA expression down-regulated by 67% and 75%, respectively, HPCs proliferation decreased by 58% and 70%. RNA-sequence demonstrated that YB-1 involved in regulation of TGF-beta signaling pathway, Hedgehog signaling pathway, Wnt signaling pathway and MAPK signaling pathway. ChIP-sequence indicated the target gene of YB-1 participated in MAPK signaling pathway, Wnt signaling pathway, Hedgehog signaling pathway. Real-time PCR and Western blotting identified down-regulation of YB-1 depressed the expression of several molecules involving in the Hedgehog signaling pathway. ChIP-PCR and luciferase reporter assay showed that YB-1 could bind to the promoter of Gli2 and modulated the transcription. CCK-8 and EdU assay indicated that Gli-1/Gli2 inhibitor GANT61 could depress HPCs proliferation up to 60%.Conclusion YB-1 can bind to the promoter of Gli2 and modulates the transcription of Gli2 further to regulate the proliferation of hepatic progenitor cells.

Key words: Y-box binding protein-1 (YB-1), Gli2, Hepatic progenitor cells, Proliferation

LI Fei, LI Bing-hang, GUO Yue-cheng, SHEN Bo, GU Tian-yi, QU Ying, ZHANG Qi-di, CAI Xiao-bo, LU Lun-gen. Y-box binding protein-1 modulates hepatic progenitor cells proliferation via Gli2[J]. Chinese Hepatolgy, 2021, 26(5): 557-562.

References

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