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Epigenetic silencing of PRSS3 provides growth and metastasis advantage for human hepatocellular carcinoma

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Abstract

Protease, serine, 3 (PRSS3), a member of the trypsin family of serine proteases, has been shown to be aberrantly expressed in several cancer types and to play important roles in tumor progression and metastasis. However, the expression and function of PRSS3 gene in hepatocellular carcinoma (HCC) remain unclear. Here we found that PRSS3 expression was decreased in human HCC cell lines and HCC surgical specimens. This was associated with intragenic methylation of PRSS3 gene. Treatment with DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine and/or histone deacetylase inhibitor trichostatin A restored PRSS3 expression in HCC cell lines. Ectopic overexpression of PRSS3 gene in HCC cell lines significantly suppressed cell proliferation and colony formation and arrested cell cycle at G1/S phase, accompanied with downregulation of cyclin D1 (CCND1)/CDK4 and cyclin E1 (CCNE1)/CDK2 complexes. Moreover, PRSS3 overexpression in HCC cells inhibited HCC cell migration and invasion with downregulation of matrix metallopeptidase 2 (MMP2). Further study showed that PRSS3 overexpression diminished the phosphorylation of mitogen-activated protein kinase/extracellular-signal-regulated kinase signaling protein, mitogen-activated protein kinase kinase 1 (MEK1)/mitogen-activated protein kinase kinase 2 (MEK2) and extracellular-signal related kinase 1 (ERK1)/extracellular-signal related kinase 2 (ERK2), in HCC cells. In contrast, knockdown of PRSS3 by small interfering RNA resulted in opposite effects on an HCC cell line SNU-387 which constitutively expresses PRSS3. These results demonstrate that downregulation of PRSS3 by intragenic hypermethylation provides growth and metastasis advantage to HCC cells. The clinical relevance of PRSS3 to human HCC was shown by the intragenic methylation of PRSS3 in HCC specimens and its association with poor tumor differentiation in patients with HCC. Thus, PRSS3 is a potential prognostic biomarker and an epigenetic target for intervention of human HCC.

Key messages

• PRSS3 is downregulated by intragenic hypermethylation in HCC.

• Epigenetic silencing of PRSS3 facilitates growth, migration, and invasion of HCC.

• PRSS3 intragenic methylation has implication in diagnosis of HCC.

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Acknowledgements

We thank Dr. Keqiang Chen for help with the data interpretation. This work was supported by the following grants: National Key Scientific Instrument Special Program of China (Grant No.2011YQ03013405); National Science Foundation of China (NSFC No.8167318, U1604281, 81230047, 81490753, 81121004, 81402345); Beijing Science Foundation of China (BJSFC No. 7171008); and Translational foundation of Chinese PLA General Hospital (2016ZHJJ-MS-GMZ). JH and JMW were also funded in part by Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E and were supported in part by the Intramural Research Program of the NCI, NIH. JH was also supported by the Intramural Research funding of Beijing Jiaotong University (S12RC00030).

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  1. Bonan Lin and Xiaomeng Zhou contributed equally to this work.

Authors and Affiliations

  1. College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, 100044, China

    Bonan Lin, Xiaomeng Zhou, Shuye Lin, Xiaoyue Wang & Jiaqiang Huang

  2. Department of Gastroenterology and Hepatology, Chinese PLA General Hospital, Beijing, 100853, China

    Meiying Zhang, Baoping Cao, Yan Dong, Shuai Yang & Mingzhou Guo

  3. Cancer and Inflammation Program, Center for Cancer Research, National Cancer Institute, Frederick, MD, 21702, USA

    Ji Ming Wang & Jiaqiang Huang

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  1. Bonan Lin
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  2. Xiaomeng Zhou
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Correspondence to Mingzhou Guo or Jiaqiang Huang.

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This study was conducted with the approval of the Institutional Review Board of the Chinese PLA General Hospital and with informed consent from patients.

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Lin, B., Zhou, X., Lin, S. et al. Epigenetic silencing of PRSS3 provides growth and metastasis advantage for human hepatocellular carcinoma. J Mol Med 95, 1237–1249 (2017). https://doi.org/10.1007/s00109-017-1578-5

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