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Cellular and Molecular Biology

Interferon regulatory factor 1 (IRF-1) downregulates Checkpoint kinase 1 (CHK1) through miR-195 to upregulate apoptosis and PD-L1 expression in Hepatocellular carcinoma (HCC) cells

British Journal of Cancer volume 125, pages 101–111 (2021)Cite this article

Subjects

Abstract

Background

CHK1 is considered an oncogene with overexpression in numerous cancers. However, CHK1 signalling regulation in hepatocellular carcinoma (HCC) remains unclear.

Methods

CHEK1 mRNA, protein, pri-miR-195 and miR-195 expression in HCC tissue was determined by qPCR, WB and IF staining assay. Survival analyses in HCC with high- and low-CHEK1 mRNA expression was performed using TCGA database. Relative luciferase activity was investigated in HCC cells transfected with p-CHEK1 3’UTR. Apoptosis was detected by TUNEL assay. NK and CD8+ T cells were analysed by flow cytometry.

Results

CHK1 is increased in human HCC tumours compared with non-cancerous liver. High CHK1 predicts worse prognosis. IFN-γ suppresses CHK1 via IRF-1 in HCC cells. The molecular mechanism of IRF-1 suppressing CHK1 is post-transcriptional by promoting miR-195 binding to CHEK1 mRNA 3’UTR, which exerts a translational blockade. Upregulated IRF-1 inhibits CHK1, which induces apoptosis of HCC cells. Likewise, CHK1 inhibition augments cellular apoptosis in HCC tumours. This effect may be a result of increased tumour NK cell infiltration. However, IRF-1 expression or CHK1 inhibition also upregulates PD-L1 expression via increased STAT3 phosphorylation.

Conclusions

IRF-1 induces miR-195 to suppress CHK1 protein expression. Both increased IRF-1 and decreased CHK1 upregulate cellular apoptosis and PD-L1 expression in HCC.

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Fig. 1: CHK1 is overexpressed in some HCC tumours.
Fig. 2: IFN-γ represses CHK1 through IRF-1 in HCC cells.
Fig. 3: IFN-γ and IRF-1 suppresses CHK1 via miR-195.
Fig. 4: Increased IRF-1 and decreased CHK1 induce apoptosis of HCC cells.
Fig. 5: Prexasertib augments cellular apoptosis of murine HCC through promoting tumour infiltrative NK cells.
Fig. 6: Increased IRF-1 and decreased CHK1 induce PD-L1 expression of HCC cells through increased phosphorylated STAT3.

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Author notes

  1. These authors contributed equally: Yihe Yan, Leting Zheng.

Authors and Affiliations

  1. Thomas E. Starzl Transplantation Institute, Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA, USA

    Yihe Yan, Leting Zheng, Qiang Du, Xiao Cui, Kun Dong, Yarong Guo & David A. Geller

  2. Department of General Surgery, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China

    Yihe Yan

  3. Department of Rheumatology and Immunology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China

    Leting Zheng

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Contributions

Y.Y. and D.A.G. designed the research. Y.Y., L.Z., Q.D., X.C., K.D. and Y.G. performed research and also analysed the data. Y.Y. and D.A.G. wrote the paper. All authors edited and approved the submission of this work.

Corresponding authors

Correspondence to Yihe Yan or David A. Geller.

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Ethics approval and consent to participate

Human tissue samples were obtained in accordance with the University of Pittsburgh Institutional Review Board (IRB) approved protocol (No. MOD08010372/PRO08010372). Animal experiments were approved by the University of Pittsburgh Institutional Animal Care and Use Committee (Protocol No. 18012053).

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Competing interests

The authors declare no competing interests.

Funding information

This work was supported by the NIH Grant (HHSN276201200017C and P30DK120531-01, D.A.G.), and the Guangxi Natural Science Foundation (2017GXNSFAA198014 and 2020GXNSFAA297008, Y.Y.).

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Yan, Y., Zheng, L., Du, Q. et al. Interferon regulatory factor 1 (IRF-1) downregulates Checkpoint kinase 1 (CHK1) through miR-195 to upregulate apoptosis and PD-L1 expression in Hepatocellular carcinoma (HCC) cells. Br J Cancer 125, 101–111 (2021). https://doi.org/10.1038/s41416-021-01337-6

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