ST6Gal-I sialyltransferase promotes chemoresistance in pancreatic ductal adenocarcinoma by abrogating gemcitabine-mediated DNA damage.

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with a poor prognosis. Gemcitabine, as a single agent or in combination therapy, remains the frontline chemotherapy despite its limited efficacy due to de novo or acquired chemoresistance. There is an acute need to decipher mechanisms underlying chemoresistance and identify new targets to improve patient outcomes. Here, we report a novel role for the ST6Gal-I sialyltransferase in gemcitabine resistance. Utilizing MiaPaCa-2 and BxPC-3 PDAC cells, we found that knockdown (KD) of ST6Gal-I expression, as well as removal of surface α2-6 sialic acids by neuraminidase, enhances gemcitabine-mediated cell death assessed via clonogenic assays and cleaved caspase 3 expression. Additionally, KD of ST6Gal-I potentiates gemcitabine-induced DNA damage as measured by comet assays and quantification of γH2AX foci. ST6Gal-I KD also alters mRNA expression of key gemcitabine metabolic genes, RRM1, RRM2, hENT1, and DCK, leading to an increased gemcitabine sensitivity ratio, an indicator of gemcitabine toxicity. Gemcitabine-resistant MiaPaCa-2 cells display higher ST6Gal-I levels than treatment-naïve cells along with a reduced gemcitabine sensitivity ratio, suggesting that chronic chemotherapy selects for clonal variants with more abundant ST6Gal-I. Finally, we examined Suit2 PDAC cells and Suit2 derivatives with enhanced metastatic potential. Intriguingly, three metastatic and chemoresistant subclones, S2-CP9, S2-LM7AA, and S2-013, exhibit up-regulated ST6Gal-I relative to parental Suit2 cells. ST6Gal-I KD in S2-013 cells increases gemcitabine-mediated DNA damage, indicating that suppressing ST6Gal-I activity sensitizes inherently resistant cells to gemcitabine. Together, these findings place ST6Gal-I as a critical player in imparting gemcitabine resistance and as a potential target to restore PDAC chemoresponse.

• Authors: Asmi Chakraborty, Kaitlyn A Dorsett, Hoa Q Trummell, Eddy S Yang, Patsy G Oliver, James A Bonner, Donald J Buchsbaum, Susan L Bellis
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 293 Issue 3 Pg. 984-994 (01 19 2018) ISSN: 1083-351X [Electronic] United States
• PMID: 29191829 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• Antimetabolites, Antineoplastic
• Equilibrative Nucleoside Transporter 1
• RNA, Messenger
• SLC29A1 protein, human
• Tumor Suppressor Proteins
• Deoxycytidine
• ribonucleotide reductase M2
• RRM1 protein, human
• Ribonucleoside Diphosphate Reductase
• Sialyltransferases
• Neuraminidase
• beta-D-Galactoside alpha 2-6-Sialyltransferase
• Gemcitabine

Topics

• Antimetabolites, Antineoplastic (pharmacology)
• Cell Line, Tumor
• Comet Assay
• DNA Damage (drug effects, genetics)
• Deoxycytidine (analogs & derivatives, pharmacology)
• Drug Resistance, Neoplasm (genetics)
• Equilibrative Nucleoside Transporter 1 (genetics)
• Humans
• Immunoblotting
• Neuraminidase (metabolism)
• Pancreatic Neoplasms (genetics)
• RNA, Messenger (genetics)
• Ribonucleoside Diphosphate Reductase (genetics)
• Sialyltransferases (genetics, metabolism)
• Tumor Suppressor Proteins (genetics)
• beta-D-Galactoside alpha 2-6-Sialyltransferase
• Gemcitabine
• Pancreatic Neoplasms