A Comprehensive Molecular Characterization of the Pancreatic Neuroendocrine Tumor Cell Lines BON-1 and QGP-1

Kim B Luley; Shauni B Biedermann; Axel Künstner; Hauke Busch; Sören Franzenburg; Jörg Schrader; Patricia Grabowski; Ulrich F Wellner; Tobias Keck; Georg Brabant; Sebastian M Schmid; Hendrik Lehnert; Hendrik Ungefroren

doi:10.3390/cancers12030691

A Comprehensive Molecular Characterization of the Pancreatic Neuroendocrine Tumor Cell Lines BON-1 and QGP-1

Cancers (Basel). 2020 Mar 14;12(3):691. doi: 10.3390/cancers12030691.

Authors

Kim B Luley¹, Shauni B Biedermann², Axel Künstner^{3

4}, Hauke Busch^{3

4}, Sören Franzenburg⁵, Jörg Schrader⁶, Patricia Grabowski⁷, Ulrich F Wellner⁸, Tobias Keck⁸, Georg Brabant², Sebastian M Schmid⁹, Hendrik Lehnert^{2

10}, Hendrik Ungefroren^{2

11}

Affiliations

¹ Clinic for Hematology and Oncology, University Hospital Schleswig-Holstein, Campus Lübeck, D-23538 Lübeck, Germany.
² First Department of Medicine, University Hospital Schleswig-Holstein, Campus Lübeck, D-23538 Lübeck, Germany.
³ Group for Medical Systems Biology, Lübeck Institute of Experimental Dermatology, University of Lübeck, D-23538 Lübeck, Germany.
⁴ Institute for Cardiogenetics, University of Lübeck, D-23538 Lübeck, Germany.
⁵ Institute for Clinical Molecular Biology, University of Kiel, Kiel, D-24105 Kiel, Germany.
⁶ First Medical Department-Gastroenterology and Hepatology, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany.
⁷ Department of Medical Immunology, Charité Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, D-13353 Berlin, Germany.
⁸ Department of Surgery, University Medical Center Schleswig-Holstein, Campus Lübeck, D-23538 Lübeck, Germany.
⁹ Institute for Endocrinology and Diabetes, University of Lübeck, D-23538 Lübeck, Germany.
¹⁰ University of Salzburg, A-5020 Salzburg, Austria.
¹¹ Clinic for General Surgery, Visceral, Thoracic, Transplantation and Pediatric Surgery, University Hospital Schleswig-Holstein, Campus Kiel, D-24105 Kiel, Germany.

Abstract

Experimental models of neuroendocrine tumor disease are scarce, with only a few existing neuroendocrine tumor cell lines of pancreatic origin (panNET). Their molecular characterization has so far focused on the neuroendocrine phenotype and cancer-related mutations, while a transcription-based assessment of their developmental origin and malignant potential is lacking. In this study, we performed immunoblotting and qPCR analysis of neuroendocrine, epithelial, developmental endocrine-related genes as well as next-generation sequencing (NGS) analysis of microRNAs (miRs) on three panNET cell lines, BON-1, QGP-1, and NT-3. All three lines displayed a neuroendocrine and epithelial phenotype; however, while insulinoma-derived NT-3 cells preferentially expressed markers of mature functional pancreatic β-cells (i.e., INS, MAFA), both BON-1 and QGP-1 displayed high expression of genes associated with immature or non-functional β/δ-cells genes (i.e., NEUROG3), or pancreatic endocrine progenitors (i.e., FOXA2). NGS-based identification of miRs in BON-1 and QGP-1 cells revealed the presence of all six members of the miR-17-92 cluster, which have been implicated in b-cell function and differentiation, but also have roles in cancer being both oncogenic or tumor suppressive. Notably, both BON-1 and QGP-1 cells expressed several miRs known to be negatively associated with epithelial-mesenchymal transition, invasion or metastasis. Moreover, both cell lines failed to exhibit migratory activity in vitro. Taken together, NT-3 cells resemble mature functional β-cells, while both BON-1 and QGP-1 are more similar to immature/non-functional pancreatic β/δ-cells or pancreatic endocrine progenitors. Based on the recent identification of three transcriptional subtypes in panNETs, NT-3 cells resemble the "islet/insulinoma tumors" (IT) subtype, while BON-1 and QGP-1 cells were tentatively classified as "metastasis-like/primary" (MLP). Our results provide a comprehensive characterization of three panNET cell lines and demonstrate their relevance as neuroendocrine tumor models.

Keywords: BON-1; EMT; NT-3; QGP-1; b-cell; microRNA; pancreatic neuroendocrine tumor.

Grants and funding

A_DE-52030-373/Ipsen Pharma