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Aspartate β-Hydroxylase expression promotes a malignant pancreatic cellular phenotype.

Dong X, Lin Q, Aihara A, Li Y, Huang CK, Chung W, Tang Q, Chen X, Carlson R, Nadolny C, Gabriel G, Olsen M, Wands JR - Oncotarget (2015)

Bottom Line: The transforming properties of ASPH depend on enzymatic activity.ASPH links PC growth factor signaling cascades to Notch activation.A small molecule inhibitor of β-hydroxylase activity was developed and found to reduce PC growth by downregulating the Notch signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical and Pharmaceutical Sciences, College of Pharmacy, The University of Rhode Island, Kingston, RI, USA.

ABSTRACT
Pancreatic cancer (PC) is one of the leading causes of cancer related deaths due to aggressive progression and metastatic spread. Aspartate β-hydroxylase (ASPH), a cell surface protein that catalyzes the hydroxylation of epidermal growth factor (EGF)-like repeats in Notch receptors and ligands, is highly overexpressed in PC. ASPH upregulation confers a malignant phenotype characterized by enhanced cell proliferation, migration, invasion and colony formation in vitro as well as PC tumor growth in vivo. The transforming properties of ASPH depend on enzymatic activity. ASPH links PC growth factor signaling cascades to Notch activation. A small molecule inhibitor of β-hydroxylase activity was developed and found to reduce PC growth by downregulating the Notch signaling pathway. These findings demonstrate the critical involvement of ASPH in PC growth and progression, provide new insight into the molecular mechanisms leading to tumor development and growth and have important therapeutic implications.

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Related in: MedlinePlus

Knockdown of ASPH by specific shRNAs in AsPC-1 cells inhibited(a) proliferation, (b) wound healing, (c) migration, (d) invasion, and (e) colony formation as indices of changes in the malignant phenotype. (f) Endogenous overexpression of ASPH was associated with upregulation of activated Notch1 and JAG2, as well as Notch signaling downstream target genes HES1, HEY1, EpCAM, CD44, c-Myc, MMP2/9, cyclin D3 and PCNA in a lentiviral transfected stable cell line expressing a nonspecific scrambled shRNA. The AsPC-1 cell lines stable transfected with ASPH specific shRNAs demonstrated downregulation of activated Notch1, JAG2, and Notch activated genes as determined by Western blot analysis. *p<0.05; **p<0.01; ***p<0.001.
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Figure 3: Knockdown of ASPH by specific shRNAs in AsPC-1 cells inhibited(a) proliferation, (b) wound healing, (c) migration, (d) invasion, and (e) colony formation as indices of changes in the malignant phenotype. (f) Endogenous overexpression of ASPH was associated with upregulation of activated Notch1 and JAG2, as well as Notch signaling downstream target genes HES1, HEY1, EpCAM, CD44, c-Myc, MMP2/9, cyclin D3 and PCNA in a lentiviral transfected stable cell line expressing a nonspecific scrambled shRNA. The AsPC-1 cell lines stable transfected with ASPH specific shRNAs demonstrated downregulation of activated Notch1, JAG2, and Notch activated genes as determined by Western blot analysis. *p<0.05; **p<0.01; ***p<0.001.

Mentions: Figure 2 panel a-e demonstrate the biological effects of ASPH overexpression on the PC cell phenotype on MIA PaCa2 as measured by proliferation, migration, invasion, as well as colony formation as an index of malignant transformation. Similar findings were evident with stable ASPH transfected Capan-1 cells (data not shown). This aggressive malignant phenotype was significantly attenuated by specific shRNAs “knockdown” of ASPH in AsPC-1 cells with high endogenous levels as demonstrated by a reduction in proliferation, migration, invasion and colony formation (Fig. 3, panel a-e).


Aspartate β-Hydroxylase expression promotes a malignant pancreatic cellular phenotype.

Dong X, Lin Q, Aihara A, Li Y, Huang CK, Chung W, Tang Q, Chen X, Carlson R, Nadolny C, Gabriel G, Olsen M, Wands JR - Oncotarget (2015)

Knockdown of ASPH by specific shRNAs in AsPC-1 cells inhibited(a) proliferation, (b) wound healing, (c) migration, (d) invasion, and (e) colony formation as indices of changes in the malignant phenotype. (f) Endogenous overexpression of ASPH was associated with upregulation of activated Notch1 and JAG2, as well as Notch signaling downstream target genes HES1, HEY1, EpCAM, CD44, c-Myc, MMP2/9, cyclin D3 and PCNA in a lentiviral transfected stable cell line expressing a nonspecific scrambled shRNA. The AsPC-1 cell lines stable transfected with ASPH specific shRNAs demonstrated downregulation of activated Notch1, JAG2, and Notch activated genes as determined by Western blot analysis. *p<0.05; **p<0.01; ***p<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359229&req=5

Figure 3: Knockdown of ASPH by specific shRNAs in AsPC-1 cells inhibited(a) proliferation, (b) wound healing, (c) migration, (d) invasion, and (e) colony formation as indices of changes in the malignant phenotype. (f) Endogenous overexpression of ASPH was associated with upregulation of activated Notch1 and JAG2, as well as Notch signaling downstream target genes HES1, HEY1, EpCAM, CD44, c-Myc, MMP2/9, cyclin D3 and PCNA in a lentiviral transfected stable cell line expressing a nonspecific scrambled shRNA. The AsPC-1 cell lines stable transfected with ASPH specific shRNAs demonstrated downregulation of activated Notch1, JAG2, and Notch activated genes as determined by Western blot analysis. *p<0.05; **p<0.01; ***p<0.001.
Mentions: Figure 2 panel a-e demonstrate the biological effects of ASPH overexpression on the PC cell phenotype on MIA PaCa2 as measured by proliferation, migration, invasion, as well as colony formation as an index of malignant transformation. Similar findings were evident with stable ASPH transfected Capan-1 cells (data not shown). This aggressive malignant phenotype was significantly attenuated by specific shRNAs “knockdown” of ASPH in AsPC-1 cells with high endogenous levels as demonstrated by a reduction in proliferation, migration, invasion and colony formation (Fig. 3, panel a-e).

Bottom Line: The transforming properties of ASPH depend on enzymatic activity.ASPH links PC growth factor signaling cascades to Notch activation.A small molecule inhibitor of β-hydroxylase activity was developed and found to reduce PC growth by downregulating the Notch signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical and Pharmaceutical Sciences, College of Pharmacy, The University of Rhode Island, Kingston, RI, USA.

ABSTRACT
Pancreatic cancer (PC) is one of the leading causes of cancer related deaths due to aggressive progression and metastatic spread. Aspartate β-hydroxylase (ASPH), a cell surface protein that catalyzes the hydroxylation of epidermal growth factor (EGF)-like repeats in Notch receptors and ligands, is highly overexpressed in PC. ASPH upregulation confers a malignant phenotype characterized by enhanced cell proliferation, migration, invasion and colony formation in vitro as well as PC tumor growth in vivo. The transforming properties of ASPH depend on enzymatic activity. ASPH links PC growth factor signaling cascades to Notch activation. A small molecule inhibitor of β-hydroxylase activity was developed and found to reduce PC growth by downregulating the Notch signaling pathway. These findings demonstrate the critical involvement of ASPH in PC growth and progression, provide new insight into the molecular mechanisms leading to tumor development and growth and have important therapeutic implications.

Show MeSH
Related in: MedlinePlus