FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes

Abstract

Background: Diabetes mellitus (DM) is a multifactorial disease orphan of a cure. Regenerative medicine has been proposed as novel strategy for DM therapy. Human fibroblast growth factor (FGF)-2b controls β-cell clusters via autocrine action, and human placental lactogen (hPL)-A increases functional β-cells. We hypothesized whether FGF-2b/hPL-A treatment induces β-cell differentiation from ductal/non-endocrine precursor(s) by modulating specific genes expression. Methods: Human pancreatic ductal-cells (PANC-1) and non-endocrine pancreatic cells were treated with FGF-2b plus hPL-A at 500 ng/mL. Cytofluorimetry and Immunofluorescence have been performed to detect expression of endocrine, ductal and acinar markers. Bromodeoxyuridine incorporation and annexin-V quantified cells proliferation and apoptosis. Insulin secretion was assessed by RIA kit, and electron microscopy analyzed islet-like clusters. Results: Increase in PANC-1 duct cells de-differentiation into islet-like aggregates was observed after FGF-2b/hPL-A treatment showing ultrastructure typical of islets-aggregates. These clusters, after stimulation with FGF-2b/hPL-A, had significant (p < 0.05) increase in insulin, C-peptide, pancreatic and duodenal homeobox 1 (PDX-1), Nkx2.2, Nkx6.1, somatostatin, glucagon, and glucose transporter 2 (Glut-2), compared with control cells. Markers of PANC-1 (Cytokeratin-19, MUC-1, CA19-9) were decreased (p < 0.05). These aggregates after treatment with FGF-2b/hPL-A significantly reduced levels of apoptosis. Conclusions: FGF-2b and hPL-A are promising candidates for regenerative therapy in DM by inducing de-differentiation of stem cells modulating pivotal endocrine genes.

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

“…Since Panc1 cells are highly invasive, these cells were used along with another non-motile PDAC cell line, Colo357, as controls in the migration assays. Panc1 cells are of ductal/exocrine origin; however, they possess some neuroendocrine features, such as expression of CgA and SSTR2 [64] and the potential to be transdifferentiated into insulin-producing cells [65][66][67][68].…”

A Comparative Endocrine Trans-Differentiation Approach to Pancreatic Ductal Adenocarcinoma Cells with Different EMT Phenotypes Identifies Quasi-Mesenchymal Tumor Cells as Those with Highest Plasticity

“…The QM cell line PANC-1 has the potential to be converted or trans-differentiated into insulin-producing cells but how this ability compares to other PDAC cells has not yet been studied. From the variety of different protocols [ 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ] we chose that of Donadel et al which involves a 5 d treatment with FGF-b and transferrin in serum-free medium [ 28 ]. Upon application of this protocol (designated P1) to PANC-1, MIA PaCa-2, COLO 357, and LüPanc1 cells we noted in PANC-1 and MIA PaCa-2 ( Figure S3A ) but not in COLO 357 and LüPanc1 (not shown) the appearance of cell aggregates resembling pancreatic islets.…”

“…TD into insulin-expressing cells used three different protocols involving either FGF-b+transferrin [ 29 ] (P1), proinflammatory cytokines [ 20 ] (P2), or IGF-1, SCF and transferrin [ 32 ] (P3). PANC-1 can be induced to form clusters that subsequently differentiate into hormone-expressing islet-like cell aggregates [ 29 ]. Various PDAC cell lines were cultured in RPMI 1640 or DMEM supplemented with 10% FBS, 1% PSG and incubated in humidified 5% CO 2 and 95% air at 37 °C.…”

“…Unfortunately, analogous studies with human pancreatic cancer cells for possible future TD therapy in PDAC patients are lacking. However, the QM PDAC cell line, PANC-1, has been successfully directed in vitro towards ductal-to-endocrine differentiation through activation of NEUROG3 [ 20 , 27 ] or induction of a β cell-like phenotype [ 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ]. The variety of different protocols employed to achieve trans-differentiation suggests that these cells are particularly plastic; however, whether this is a consequence of their QM signature remains elusive as equivalent data for other PDAC-derived cell lines are not available.…”

Placental Lactogen as a Marker of Maternal Obesity, Diabetes, and Fetal Growth Abnormalities: Current Knowledge and Clinical Perspectives

“…The exposure of human pancreatic ductal cells (PANC-1) to a mixture of PL and human fibroblast growth factor-2b could effectively induce their de-differentiation to active islet β cells. After treatment, PANC-1 changed their ultrastructure to that typical of islets-aggregates, showing a significantly increased production of insulin and C peptide, which could be a promising new approach to diabetes treatment [59]. Similar to rodent models, PL stimulates the maternal prolactin receptors in human pancreatic β cells, promoting their adaptations to increased insulin requirements during physiological gestation [41,60].…”