Inhibition of Checkpoint Kinase 1 (CHK1) Upregulates Interferon Regulatory Factor 1 (IRF1) to Promote Apoptosis and Activate Anti-Tumor Immunity via MICA in Hepatocellular Carcinoma (HCC)

Simple Summary The communication between the DNA damage response pathway and the tumor microenvironment of hepatocellular carcinoma (HCC) has attracted more attention. In this study, high CHK1 expression in HCC tissue was associated with advanced tumor stage and poor prognosis in patients. CHK1 inhibition and cisplatin induced cellular apoptosis via DNA damage in human HCC cells. CHK1 directly bound IRF1 and exerted a proteolytic effect in HCC cells induced by DNA damage. DNA damage induced MICA expression via IRF1 at the transcriptional level in HCC cells. MICA expression was positively correlated with NK cells and CD8+T cell infiltration in human HCC. This study provided new insight into the molecular mechanisms regulating HCC signaling with cross-talk between the IRF1 and CHK1 pathways. Background: CHK1 is considered a key cell cycle checkpoint kinase in DNA damage response (DDR) pathway to communicate with several signaling pathways involved in the tumor microenvironment (TME) in numerous cancers. However, the mechanism of CHK1 signaling regulating TME in hepatocellular carcinoma (HCC) remains unclear. Methods: CHK1 expression in HCC tissue was determined by IHC staining assay. DNA damage and apoptosis in HCC cells induced by cisplatin or CHK1 inhibition were detected by WB and flow cytometry. The interaction of CHK1 and IRF1 was analyzed by single-cell RNA-sequence, WB, and immunoprecipitation assay. The mechanism of IRF1 regulating MICA was investigated by ChIP-qPCR. Results: CHK1 expression is upregulated in human HCC tumors compared to the background liver. High CHK1 mRNA level predicts advanced tumor stage and worse prognosis. Cisplatin and CHK1 inhibition augment cellular DNA damage and apoptosis. Overexpressed CHK1 suppresses IRF1 expression through proteolysis. Furthermore, single-cell RNA-sequence analyses confirmed that MICA expression positively correlated with IRF1 in HCC cells. Immunoprecipitation assay showed the binding between CHK1 and IRF1. Cisplatin and CHK1 inhibition upregulate MICA expression through IRF1-mediated transcriptional effects. A novel specific cis-acting IRF response element was identified at -1756 bp in the MICA promoter region that bound IRF1 to induce MICA gene transcription. MICA may increase NK cell and CD8+T cell infiltration in HCC. Conclusions: DNA damage regulates the interaction of CHK1 and IRF1 to activate anti-tumor immunity via the IRF1-MICA pathway in HCC.

Automated summary

This study reveals the relationship between CHK1 expression and the tumor microenvironment in hepatocellular carcinoma (HCC). High CHK1 expression is associated with advanced tumor stage and poor prognosis. CHK1 inhibition and cisplatin induce DNA damage and apoptosis in HCC cells. CHK1 directly interacts with IRF1 to regulate MICA expression, which is correlated with NK cell and CD8+T cell infiltration in HCC.