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 β-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.
Highlights
Preclinical models of neuroendocrine tumors (NET) currently encompass only a limited number of human NET cell lines and mouse models
To verify or dismiss such a scenario, we evaluated in be the result of absent (BON), QGP, and NT-3 cells expression of Insulin, Glucagon, and SST by Quantitative Real-Time RT-PCR (qPCR) analysis
We showed that the BON and QGP cell lines display a neuroendocrine and well-differentiated phenotype, but, in contrast to the NT-3 line, do not exhibit a gene signature characteristic of mature/functional pancreatic islet/β-cells
Summary
Preclinical models of neuroendocrine tumors (NET) currently encompass only a limited number of human NET cell lines and mouse models. Their mutation patterns, and proliferation rates seem distinct from well-differentiated NETs in patients [5,6,7] and might not adequately reflect the tumor biology of well-differentiated NETs. for panNET, studies in BON and QGP cells have raised questions regarding their relevance as models due to the absence of mutations in MEN1 [5,6,7] or low SSTR expression [8]. For panNET, studies in BON and QGP cells have raised questions regarding their relevance as models due to the absence of mutations in MEN1 [5,6,7] or low SSTR expression [8] Together, this emphasizes the need for careful reevaluation and further characterization of the existing cell lines. One aim of the present study was; to confirm the authenticity of the BON and QGP cell lines with respect to their neuroendocrine and epithelial phenotype, developmental origin, and propensity for cell motility in vitro
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