Aspartate β-hydroxylase modulates cellular senescence through glycogen synthase kinase 3β in hepatocellular carcinoma

Yoshifumi Iwagami, Chiung Kuei Huang, Mark J. Olsen, John Michael Thomas, Grace Jang, Miran Kim, Qiushi Lin, Rolf I. Carlson, Carl Wagner, Xiaoqun Dong, Jack R. Wands

Research output: Contribution to journalArticle

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Abstract

Aspartate β-hydroxylase (ASPH) is an enzyme overexpressed in human hepatocellular carcinoma (HCC) tumors that participates in the malignant transformation process. We determined if ASPH was a therapeutic target by exerting effects on cellular senescence to retard HCC progression. ASPH knockdown or knockout was achieved by short hairpin RNAs or the CRISPR/Cas9 system, respectively, whereas enzymatic inhibition was rendered by a potent second-generation small molecule inhibitor of ASPH. Alterations of cell proliferation, colony formation, and cellular senescence were evaluated in human HCC cell lines. The potential mechanisms for activating cellular senescence were explored using murine subcutaneous and orthotopic xenograft models. Inhibition of ASPH expression and enzymatic activity significantly reduced cell proliferation and colony formation but induced tumor cell senescence. Following inhibition of ASPH activity, phosphorylation of glycogen synthase kinase 3β and p16 expression were increased to promote senescence, whereas cyclin D1 and proliferating cell nuclear antigen were decreased to reduce cell proliferation. The mechanisms involved demonstrate that ASPH binds to glycogen synthase kinase 3β and inhibits its subsequent interactions with protein kinase B and p38 upstream kinases as shown by coimmunoprecipitation. In vivo experiments demonstrated that small molecule inhibitor treatment of HCC bearing mice resulted in significant dose-dependent reduced tumor growth, induced phosphorylation of glycogen synthase kinase 3β, enhanced p16 expression in tumor cells, and promoted cellular senescence. Conclusions: We have identified a new mechanism that promotes HCC growth and progression by modulating senescence of tumor cells; these findings suggest that ASPH enzymatic activity is a novel therapeutic target for HCC. (Hepatology 2016;63:1213-1226)

Original languageEnglish (US)
Pages (from-to)1213-1226
Number of pages14
JournalHepatology
Volume63
Issue number4
DOIs
StatePublished - Apr 1 2016

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Glycogen Synthase Kinase 3
Cell Aging
Mixed Function Oxygenases
Aspartic Acid
Hepatocellular Carcinoma
Cell Proliferation
Neoplasms
Clustered Regularly Interspaced Short Palindromic Repeats
Phosphorylation
Proto-Oncogene Proteins c-akt
Cyclin D1
Proliferating Cell Nuclear Antigen
Gastroenterology
Growth
Heterografts
Small Interfering RNA
Phosphotransferases
Cell Line

ASJC Scopus subject areas

  • Medicine(all)
  • Hepatology

Cite this

Iwagami, Y., Huang, C. K., Olsen, M. J., Thomas, J. M., Jang, G., Kim, M., ... Wands, J. R. (2016). Aspartate β-hydroxylase modulates cellular senescence through glycogen synthase kinase 3β in hepatocellular carcinoma. Hepatology, 63(4), 1213-1226. https://doi.org/10.1002/hep.28411

Aspartate β-hydroxylase modulates cellular senescence through glycogen synthase kinase 3β in hepatocellular carcinoma. / Iwagami, Yoshifumi; Huang, Chiung Kuei; Olsen, Mark J.; Thomas, John Michael; Jang, Grace; Kim, Miran; Lin, Qiushi; Carlson, Rolf I.; Wagner, Carl; Dong, Xiaoqun; Wands, Jack R.

In: Hepatology, Vol. 63, No. 4, 01.04.2016, p. 1213-1226.

Research output: Contribution to journalArticle

Iwagami, Y, Huang, CK, Olsen, MJ, Thomas, JM, Jang, G, Kim, M, Lin, Q, Carlson, RI, Wagner, C, Dong, X & Wands, JR 2016, 'Aspartate β-hydroxylase modulates cellular senescence through glycogen synthase kinase 3β in hepatocellular carcinoma', Hepatology, vol. 63, no. 4, pp. 1213-1226. https://doi.org/10.1002/hep.28411
Iwagami, Yoshifumi ; Huang, Chiung Kuei ; Olsen, Mark J. ; Thomas, John Michael ; Jang, Grace ; Kim, Miran ; Lin, Qiushi ; Carlson, Rolf I. ; Wagner, Carl ; Dong, Xiaoqun ; Wands, Jack R. / Aspartate β-hydroxylase modulates cellular senescence through glycogen synthase kinase 3β in hepatocellular carcinoma. In: Hepatology. 2016 ; Vol. 63, No. 4. pp. 1213-1226.
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