Novel Pancreatic Cancer Cell Lines Derived from Genetically Engineered Mouse Models of Spontaneous Pancreatic Adenocarcinoma: Applications in Diagnosis and Therapy

Abstract

Pancreatic cancer (PC) remains one of the most lethal human malignancies with poor prognosis. Despite all advances in preclinical research, there have not been significant translation of novel therapies into the clinics. The development of genetically engineered mouse (GEM) models that produce spontaneous pancreatic adenocarcinoma (PDAC) have increased our understanding of the pathogenesis of the disease. Although these PDAC mouse models are ideal for studying potential therapies and specific genetic mutations, there is a need for developing syngeneic cell lines from these models. In this study, we describe the successful establishment and characterization of three cell lines derived from two (PDAC) mouse models. The cell line UN-KC-6141 was derived from a pancreatic tumor of a KrasG12D;Pdx1-Cre (KC) mouse at 50 weeks of age, whereas UN-KPC-960 and UN-KPC-961 cell lines were derived from pancreatic tumors of KrasG12D;Trp53R172H;Pdx1-Cre (KPC) mice at 17 weeks of age. The cancer mutations of these parent mice carried over to the daughter cell lines (i.e. KrasG12D mutation was observed in all three cell lines while Trp53 mutation was observed only in KPC cell lines). The cell lines showed typical cobblestone epithelial morphology in culture, and unlike the previously established mouse PDAC cell line Panc02, expressed the ductal marker CK19. Furthermore, these cell lines expressed the epithelial-mesenchymal markers E-cadherin and N-cadherin, and also, Muc1 and Muc4 mucins. In addition, these cell lines were resistant to the chemotherapeutic drug Gemcitabine. Their implantation in vivo produced subcutaneous as well as tumors in the pancreas (orthotopic). The genetic mutations in these cell lines mimic the genetic compendium of human PDAC, which make them valuable models with a high potential of translational relevance for examining diagnostic markers and therapeutic drugs.

NF-κB regulates GDF-15 to suppress macrophage surveillance during early tumor development

“…Binding of p65 also corresponded to enhanced occupancy of phosphorylated RNA polymerase II (pPol II, which is phosphorylated on serine 2) ( Figure 4F), suggesting that the binding of p65 to exon 2 of Gdf-15 promotes active transcription. To further substantiate these findings, we constructed a luciferase reporter plasmid containing the NF-κB-binding element from exon 2 of Gdf- 15. Results showed that GDF-15 reporter activity was enhanced in p65 +/+ Ras MEFs and in cells stimulated by TNF (Figure 4, G and H).…”

“…Therefore, we expanded our analysis generating additional Gdf- 15- (15,16) that also stably expressed a luciferase reporter gene that was used for bioluminescence imaging (17) (Figure 3A). KPC control or KPC Gdf-15-knockdown clones were then orthotopically injected into the tail of the pancreas of C57BL/6 albino mice, and tumor development was visualized.…”

Combination therapy with BPTES nanoparticles and metformin targets the metabolic heterogeneity of pancreatic cancer

“…PDAC cells were maintained in DMEM with 10% (vol/vol) FBS and 1% penicillin/streptomycin in a 5% (vol/vol) CO 2 and 95% (vol/vol) air incubator. All cell lines (P8, A6L, A32, P198, E3, P215, P10, JD13D) were generated from PDAC tumors as previously described (46)(47)(48) and kindly provided by Anirban Maitra (MD Anderson Cancer Center, Houston). To determine glutamine dependence, cells were grown with or without glutamine in DMEM with 10% (vol/vol) dialyzed FBS and 1% penicillin/streptomycin.…”

Tumor-penetrating peptide enhances transcytosis of silicasome-based chemotherapy for pancreatic cancer

“…This stringent PDAC tumor model mimics human PDAC for oncogene expression, growth characteristics, metastasis, histological features, and development of a dysplastic stroma (28,29). Figure 1B summarizes the details of the tumor growth characteristics and metastasis as seen during animal autopsy and IVIS imaging.…”

“…Pdx1-Cre mouse) (27)(28)(29). In addition, silicasomes also provide major toxicity reduction in the gastrointestinal tract, liver, and bone marrow compared with the liposomal equivalent (27).…”