HDAC3-dependent transcriptional repression of FOXA2 regulates FTO/m6A/MYC signaling to contribute to the development of gastric cancer

Abstract

As one of the deadliest malignancies, gastric cancer (GC) is often accompanied by a low 5-year survival following initial diagnosis, which accounts for a substantial proportion of cancer-related deaths each year worldwide. Altered epigenetic modifications of cancer oncogenes and tumor suppressor genes emerge as novel mechanisms have been implicated the pathogenesis of GC. In the current study, we aim to elucidate whether histone deacetylase 3 (HDAC3) exerts oncogenic role in GC, and investigate the possible mechanism. Initially, we collected 64 paired cancerous and noncancerous tissues surgically resected from GC patients. Positive expression of HDAC3, FTO, and MYC in the tissues was measured using Immunohistochemistry. Meanwhile, GC cell line BGC-823/AGS was selected and treated with lentivirus vectors for alteration of HDAC3, FTO, or FOXA2 expressions, followed by detection on mRNA and protein levels of HDAC3, FOXA2, FTO, and MYC using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot assays. The results demonstrated that the expressions of HDAC3, FTO and MYC were upregulated, while FOXA2 expression was downregulated in GC tissues and cells. After that, the cell viability, migration, and invasion of GC cells were assessed by CCK-8 and Transwell assays, revealing that HDAC3 accelerated GC cell viability, migration and invasion by degrading FOXA2. Subsequently, the binding relationship among HDAC3, FOXA2, FTO, and MYC was assessed by assays of immunoprecipitation, dual-luciferase reporter gene, and chromatin immunoprecipitation assay. Methylation of m6A mRNA in GC cells was detected via gene-specific m6A qPCR and dot-blot assays. The transcription factor FOXA2 was found to bind to the FTO gene promoter and decreased its expression, while FTO stabilized MYC mRNA by reducing m6A methylation of MYC in GC cells. In addition, HDAC3 was observed to maintain the FTO/m6A/MYC signaling and regulated GC progression, which was also supported by in vivo animal study data of GC cell tumorigenesis in nude mice. These key observations uncover the tumor-initiating activities of HDAC3 in GC through its regulation on FOXA2-mediated FTO/m6A/MYC axis, highlighting the potential of therapeutically targeting epigenetic modifications to combat GC.

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Fig. 1: Expression pattern of FTO in GC by web-available databases.
Fig. 2: Expression of HDAC3, FTO, and MYC proteins in GC and adjacent normal tissues (n = 64) (×400).
Fig. 3: HDAC3 is upregulated in GC and promotes GC cell proliferation, migration, and invasion in vitro.
Fig. 4: HDAC3 enhances GC cell proliferation, migration, and invasion via downregulation of FOXA2.
Fig. 5: FOXA2 could specifically bind to FTO promoter and inhibit its transcriptional activity to suppress FTO expression in GC cells.
Fig. 6: FTO stabilizes MYC mRNA by reducing m6A methylation of MYC in GC cells.
Fig. 7: HDAC3 targets FTO/m6A/MYC signaling and regulates GC cells in vitro.
Fig. 8: Depletion of HDAC3 inhibits tumor growth via FOXA2/FTO/ MYC axis.
Fig. 9: The potential molecular mechanism for HDAC3/FOXA2/FTO/MYC signaling axis in the development of gastric cancer.

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Acknowledgements

We acknowledge anyone who contributed toward the article who does not meet the criteria for authorship including anyone who provided professional writing services or materials.

Funding

This work was supported by the National Natural Science Foundation of China (No. 81672877; No. 81502073).

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Correspondence to Xiaofeng Jiang or Haiying Zhao.

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Yang, Z., Jiang, X., Zhang, Z. et al. HDAC3-dependent transcriptional repression of FOXA2 regulates FTO/m6A/MYC signaling to contribute to the development of gastric cancer. Cancer Gene Ther (2020). https://doi.org/10.1038/s41417-020-0193-8

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