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RUNX3-mediated transcriptional inhibition of Akt suppresses tumorigenesis of human gastric cancer cells

Oncogene volume 31, pages 4302–4316 (2012)Cite this article

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Abstract

Activation of Akt signaling pathway has been suggested involving in chemoresistance, metastasis and tumorigenesis of gastric cancer. However, the mechanism of Akt regulation in gastric cancer is not fully understood. RUNX3, which was first identified as a transcription factor, suppresses gastric tumorigenesis through regulating expression of target genes. Here, we found that restoration of RUNX3 significantly downregulates the protein and mRNA expression of Akt1 in gastric cancer cell lines, AGS and SNU-1. Knockdown of RUNX3 upregulates protein and mRNA expression of Akt1 in normal gastric epithelial cell line, GES-1. The negative correlation of RUNX3 and Akt expression and downstream β-catenin/cyclin D1 effectors was further confirmed in AGS and GES-1 cell lines, as well as clinical specimens of gastric cancer. We identified two RUNX3-binding sites in Akt1 promoter and the binding of RUNX3 on Akt1 promoter significantly inhibits Akt1 expression. The RUNX3-mediated inhibition of Akt1 caused β-catenin protein degradation and then cyclin D1 downregulation. Restoration of cyclin D1 reverses cell growth inhibition and G1 phase arrest induced by RUNX3 in gastric cancer cells. Our results show that loss of RUNX3 expression can enhance the Akt1-mediated signaling pathway and promote the tumorigenesis process in human gastric cancer.

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Acknowledgements

The study was supported by grants from the National Science Council of Taiwan (NSC 97-2311-B-006-003-MY3 and NSC 99-2627-B-006-021), National Cheng Kung University Hospital of Taiwan (NCKUH-9803004 and NCKUH-9903051) and Chi Mei Medical Center of Taiwan (CMNCKU9901). We acknowledge Dr Xiang Hong Zhang from Hebei Medical University in China for kindly providing GES-1 cell line, and Dr Ching-Shih Chen from Ohio State University for kindly providing Akt, myr-Akt and DN-Akt constructs.

Author contributions: F-CL, Y-PL, MH and P-JL designed the research. C-HLai carried out part of the experiments. F-CL and Y-PL analyzed the data. C-HLai and C-HW helped to design cloning strategies. H-CC, C-HLai and C-HW carried out the immunohistochemistry. JQC provided Akt promoter construct. F-CL, Y-PL, C-HLee, Y-CL, H-YW and P-JL contributed to extra technical assistance. P-IH and Y-SS provided clinical specimens. MH helped immunohistochemistry scoring for specimens. F-CL, Y-PL and P-JL organized the figures and wrote the paper. All authors participated in the interpretation of the data and production of the final paper.

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  1. F-C Lin and Y-P Liu: These authors contribute equally to this work.

Authors and Affiliations

  1. Institute of Clinical Medicine, National Cheng Kung University, Medical College, Tainan, Taiwan

    F-C Lin, Y-P Liu, C-H Lai, Y-S Shan, H-C Cheng, Y-C Lee, H-Y Wang & P-J Lu

  2. Department of Radiation Oncology, National Cheng Kung University Hospital, Tainan, Taiwan

    F-C Lin

  3. Genomics Research Center, Academic Sinica, Taipei, Taiwan

    Y-P Liu, C-H Lee & M Hsiao

  4. Department of Surgery, National Cheng Kung University Hospital, Tainan, Taiwan

    Y-S Shan

  5. Department of Internal Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan

    P-I Hsu

  6. Department of Internal Medicine, National Yang-Ming University, Taipei, Taiwan

    P-I Hsu

  7. Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan

    C-H Wang

  8. Department of Molecular Oncology, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    J Q Cheng

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Lin, FC., Liu, YP., Lai, CH. et al. RUNX3-mediated transcriptional inhibition of Akt suppresses tumorigenesis of human gastric cancer cells. Oncogene 31, 4302–4316 (2012). https://doi.org/10.1038/onc.2011.596

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