Pancreatic Cancer Cell Lines PANC-1 Research Articles

MicroRNA miR-374, a potential radiosensitizer for carbon ion beam radiotherapy.

In this study, we compared the microRNA(miRNA) profiles of a control and X-ray- and carbon ion beam-resistant cells to identify miRNAs that can be used as radiosensitizers and biomarkers. Mouse squamous cell carcinoma line NR-S1, its X-ray-resistant derivative X60, and its carbon ion beam‑resistant derivative C30 were subjected to miRNA microarray analysis. Expression of miRNAs shown to be upregulated or downregulated in the microarray analysis was confirmed by qRT-PCR. Downregulated miRNAs were overexpressed in human pancreatic cancer cell lines PANC1 and MIA PaCa-2, and the resulting cells were tested for radiosensitivity using colony-forming and sphere-forming assays. Of 1,265miRNAs analyzed, 4were downregulated and11 were upregulated in X-ray-resistant and carbon ion beam-resistant cells. Two of the downregulated miRNAs, miR-196 and miR-374, were selected for overexpression in PANC1 and MIA PaCa-2 cells. Overexpression of miR-374 sensitized PANC-1 and MIA PaCa-2 cells toward carbon ion beam radiation. miRNA miR-374 has the potential to be a new radiosensitizer for carbon ion beam radiotherapy and a new biomarker to determine the optimal treatment for cancer.

A Size-Selective Intracellular Delivery Platform.

Identifying and separating a subpopulation of cells from a heterogeneous mixture are essential elements of biological research. Current approaches require detailed knowledge of unique cell surface properties of the target cell population. A method is described that exploits size differences of cells to facilitate selective intracellular delivery using a high throughput microfluidic device. Cells traversing a constriction within this device undergo a transient disruption of the cell membrane that allows for cytoplasmic delivery of cargo. Unique constriction widths allow for optimization of delivery to cells of different sizes. For example, a 4 μm wide constriction is effective for delivery of cargo to primary human T-cells that have an average diameter of 6.7 μm. In contrast, a 6 or 7 μm wide constriction is best for large pancreatic cancer cell lines BxPc3 (10.8 μm) and PANC-1 (12.3 μm). These small differences in cell diameter are sufficient to allow for selective delivery of cargo to pancreatic cancer cells within a heterogeneous mixture containing T-cells. The application of this approach is demonstrated by selectively delivering dextran-conjugated fluorophores to circulating tumor cells in patient blood allowing for their subsequent isolation and genomic characterization.

Cell-cycle-controlled radiation therapy was effective for treating a murine malignant melanoma cell line in vitro and in vivo.

Radiotherapy is a commonly used regimen for treating various types of intractable cancers, although the effects depend on the cell cycle of the targeted cancer cell lines, and for irradiation purposes it is therefore critical to establish a protocol for controlling the cell cycle. Here, we showed that a common murine melanoma cell line B16BL6 was more vulnerable to irradiation during the early S phase, and that synchronisation of the cell cycle greatly increased the therapeutic effects of radiotherapy. Cell-sorting experiments, according to cell-cycle phase, using B16BL6 cells demonstrated that cells in the early S phase were the most susceptible to radiotherapy. Gemcitabine, a clinically utilised anti-cancer drug, induced cell-cycle arrest during the early S phase in B16BL6 cells, and thus a synergistic therapeutic effect was observed when irradiation was administered at the right time. Human pancreatic cancer cell line PANC-1 exhibited similar properties to B16BL6 in terms of its radiosensitivity during the S/G2/M phase and also demonstrated a synergistic effect of cell cycle synchronisation. These results show the importance of cell-cycle control in the application of irradiation and suggest a suitable time interval between chemotherapy and radiotherapy, as well as providing useful information for treating intractable cancer.

Leiodermatolide, a novel marine natural product, has potent cytotoxic and antimitotic activity against cancer cells, appears to affect microtubule dynamics, and exhibits antitumor activity.

Pancreatic cancer, the fourth leading cause of cancer death in the United States, has a negative prognosis because metastasis occurs before symptoms manifest. Leiodermatolide, a polyketide macrolide with antimitotic activity isolated from a deep water sponge of the genus Leiodermatium, exhibits potent and selective cytotoxicity toward the pancreatic cancer cell lines AsPC-1, PANC-1, BxPC-3, and MIA PaCa-2, and potent cytotoxicity against skin, breast and colon cancer cell lines. Induction of apoptosis by leiodermatolide was confirmed in the AsPC-1, BxPC-3 and MIA PaCa-2 cells. Leiodermatolide induces cell cycle arrest but has no effects on in vitro polymerization or depolymerization of tubulin alone, while it enhances polymerization of tubulin containing microtubule associated proteins (MAPs). Observations through confocal microscopy show that leiodermatolide, at low concentrations, causes minimal effects on polymerization or depolymerization of the microtubule network in interphase cells, but disruption of spindle formation in mitotic cells. At higher concentrations, depolymerization of the microtubule network is observed. Visualization of the growing microtubule in HeLa cells expressing GFP-tagged plus end binding protein EB-1 showed that leiodermatolide stopped the polymerization of tubulin. These results suggest that leiodermatolide may affect tubulin dynamics without directly interacting with tubulin and hint at a unique mechanism of action. In a mouse model of metastatic pancreatic cancer, leiodermatolide exhibited significant tumor reduction when compared to gemcitabine and controls. The antitumor activities of leiodermatolide, as well as the proven utility of antimitotic compounds against cancer, make leiodermatolide an interesting compound with potential chemotherapeutic effects that may merit further research.

Interleukin-32α inactivates JAK2/STAT3 signaling and reverses interleukin-6-induced epithelial-mesenchymal transition, invasion, and metastasis in pancreatic cancer cells.

Interleukin (IL)-32 is a newly discovered cytokine that has multifaceted roles in inflammatory bowel disease, cancer, and autoimmune diseases and participates in cell apoptosis, cancer cell growth inhibition, accentuation of inflammation, and angiogenesis. Here, we investigated the potential effects of IL-32α on epithelial–mesenchymal transition, metastasis, and invasion, and the JAK2/STAT3 signaling pathway in pancreatic cancer cells. The human pancreatic cancer cell lines PANC-1 and SW1990 were used. Epithelial–mesenchymal transition-related markers, including E-cadherin, N-cadherin, Vimentin, Snail, and Zeb1, as well as extracellular matrix metalloproteinases (MMPs), including MMP2, MMP7, and MMP9, were detected by immunofluorescence, Western blotting, and real-time polymerase chain reaction. The activation of JAK2/STAT3 signaling proteins was detected by Western blotting. Wound healing assays, real-time polymerase chain reaction, and Western blotting were performed to assess cell migration and invasion. The effects of IL-32α on the IL-6-induced activation of JAK2/STAT3 were also evaluated. In vitro, we found that IL-32α inhibits the expressions of the related markers N-cadherin, Vimentin, Snail, and Zeb1, as well as JAK2/STAT3 proteins, in a dose-dependent manner in pancreatic cancer cell lines. Furthermore, E-cadherin expression was increased significantly after IL-32α treatment. IL-32α downregulated the expression of MMPs, including MMP2, MMP7, and MMP9, and decreased wound healing in pancreatic cancer cells. These consistent changes were also found in IL-6-induced pancreatic cancer cells following IL-32α treatment. This study showed that reversion of epithelial–mesenchymal transition, inhibition of invasiveness and metastasis, and activation of the JAK2/STAT3 signaling pathway could be achieved through the application of exogenous IL-32α.

The hepatitis B virus X protein promotes pancreatic cancer through modulation of the PI3K/AKT signaling pathway

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive and lethal cancer, with poor outcomes. Infection with the hepatitis B virus (HBV) may be associated as a worse prognosis for PDAC patients; however, the mechanisms involved in this process are unclear. We evaluated whether HBV infection leads to PDAC with a more aggressive phenotype, and attempted to elucidate the mechanisms involved. Clinicopathological data and outcomes from 64 patients with PDAC were collected and compared between serum HBsAg+ and HBsAg− patients. Furthermore, we examined the effects of the HBV X protein (HBx) on proliferation and migration of the pancreatic cancer cell lines PANC-1 and SW1990. We investigated expression changes of over 500 proteins by protein array analysis and identified several HBV- and PDAC-related candidates, which were further validated by immunoblotting and enzyme-linked immunosorbent assay. No differences in clinicopathological features were observed between HBsAg+ and HBsAg− patients; however, HBsAg+ patients had a shorter median survival time (8 vs. 13 months), although the differences were not significant. HBV DNA was detected in clinical specimens, even in PDAC patients considered “HBV-free”, potentially due to occult infection. HBx expression significantly enhanced cellular proliferation and migration and induced an epithelial-mesenchymal transition phenotype. Expression of ErbB4 and TGF-α was increased in parallel with HBx expression, and several downstream pathways including PI3K/AKT, MAPK, and ERK were upregulated. Inhibition of the PI3K/AKT pathway reversed the effects of HBx in PDAC cell lines. HBx may, therefore, contribute to the progression of PDAC through modulation of these pathways.

A microfluidic multiwell chip for enzyme-free detection of mRNA from few cells

Isogenic cell populations possess heterogeneous gene expression patterns. Most methods for mRNA expression analysis start with the reverse transcription of mRNA into cDNA, a process that can introduce strong signal variations not related to the actual mRNA levels. Miniaturized lab-on-a-chip systems offer properties – e.g. low sample dilution, low contamination – that enable new reaction schemes for molecular analyses. To enable transcription-free mRNA expression analysis of few single cells, a one-step cell lysis, target labelling and hybridisation approach as well as a corresponding passive multiwell chip with a volume of 25.5 nL/well were developed. The method enabled the parallel analysis of up to 96 samples and 6 target genes per sample. Preceding light microscopy of the living cells allowed correlating mRNA levels and cell number. As a proof-of-principle, the pancreatic cancer cell line Panc-1 was investigated for expression heterogeneity of a reference gene plus 5 genes reported to be overexpressed in cancer stem cells (CSCs). A good correlation (r(51)=0.739, p<0.001; rs(51)=0.744, p<0.001) between the cell number per well and the number of detected reference gene mRNA confirmed the proper function of the device. Moreover, a heterogeneous expression of the CSC-associated target genes was found which matched well with reports on the presence of CSCs in the Panc-1 cell line.

Investigation of juglone effects on metastasis and angiogenesis in pancreatic cancer cells

Juglone, a natural component, is shown to have cytotoxic and apoptotic effects in several cancer cell lines. However, little is known about its effects on invasion and metastasis. In this study, we aimed to determine the antimetastatic effect of juglone in the BxPC-3 and PANC-1 pancreatic cancer cell lines. Cytotoxic effect of juglone was evaluated by using MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) test. The cells were treated with juglone at <IC50 doses (5, 10, 15 and 20μM ) for 24h. After the cell adhesion and invasion analysis, expression profiles of the MMP-2, MMP-9 and Phactr-1 genes were determined by qPCR. The IC50 dose of juglone was found to be 21.05μM in the BxPC-3 cell line and 21.25μM in the PANC-1 cell line for 24h. According to the cell adhesion and invasion analysis, treatment of juglone for 24h reduced the adhesion and invasion features of pancreatic cancer cells. A significant reduction of MMP-2, MMP-9 and Phactr-1 expressions was observed in pancreatic cancer cells after the treatment of juglone at <IC50 doses. By this study, it has been shown for the first time that juglone inhibits cell invasion and metastasis in pancreatic cancer line and can be evaluated as an effective anticancer agent in pancreatic cancer.

Study on effect and mechanism of ellagic acid on the proliferation and apoptosis of pancreatic cancer cell line PANC-1

Objective To investigate the effect of ellagic acid on the proliferation and apoptosis of pancreatic cell line PANC-1 in vitro and explore possible mechanism of ellagic acid's effect on PANC-1 cells. Methods PANC-1 cells were treated with ellagic acid at concentrations of 0, 2, 4, 6, 8 μg/ml for 24, 48 and 72 h respectively. Inverted Microscope showed cells' morphological changes. The cell proliferation was determined by methyl thiazolyl tetrazolium (MTT) assay. When the cells were treated with ellagic acid at concentrations of 0, 2, 4, 6 μg/ml for 48 h, the morphological changes of apoptotic cells were observed by cell staining, apoptotic rate and cell cycle were detected by flow cytometry, the protein expression of cyclo-oxgenase 2 (COX-2) and nuclear factor-κB (NF-κB) were measured by Western blotting. Results Ellagic acid could significantly inhibited the growth of PANC-1 cells in a concentration-dependent and time-dependent manner. The difference among these groups were statistically significant(P<0.05). After treatment with ellagic acid, apoptosis of PANC-1 cells was seen. Apoptosis rates in the different concentration groups were respectively (11.18±0.99)%, (20.70±2.32)%, (39.68±0.69)%, the apoptosis rate of control group [(0.62±0.23)%] compared with statistically significant difference among the different concentration groups (P<0.05). The percentages of G0/G1 phase cells were respectively(48.98±2.36)%, (60.80±1.40)%, (72.82±0.59)%, the percentage of control group [(42.78±0.74)%] compared with statistically significant difference among the different concentration groups(P<0.05). The protein expression of COX-2 and NF-κB measured by Western blotting were significantly down-regulated dose-dependently. Conclusion Ellagic acid can inhibit the proliferation of pancreatic cancer cell line PANC-1 and induce apoptosis. The mechanism may depend on arresting cell cycle and decreasing protein expression of COX-2 and NF-κB. Key words: Ellagic acid; Pancreatic neoplasm; Proliferation; Apoptosis; Cyclo-oxgenase 2; Nuclear factor-κB

Synthesis of new tricyclic thiolactams as potent antitumor agent for pancreatic cancer

We synthesized the novel tricyclic thiolactams 2a–d, 3d–k, having a benzyl or substituted benzyl substituent on the nitrogen of indole subunit, and their preferential cytotoxicity under both nutrient-deprived medium (NDM) and Dulbecco’s modified Eagle’s medium (DMEM) was evaluated against a human pancreatic cancer cell line PANC-1. Among the tested compounds, the 4′-hydroxy derivative 3d showed the most potent cytotoxicity in NDM (PC50 1.68μM) although the moderate preferential cytotoxicity (PC50 1.68μM in NDM vs PC50 20μM in DMEM). The 3′-hydroxy derivative 3e exhibited the most preferential cytotoxicity (PC50 1.96μM in NDM vs less than 50% inhibition at 30μM in DMEM). The benzyl 2a and halogenated benzyl derivatives 2b,c showed no cytotoxicity in NDM. In addition, the indole (10, PC50 173.7μM), lactone (11, PC50 131.7μM), and lactam (12, PC50 44.8μM) derivatives showed week or moderate cytotoxicity in NDM. These results indicated that the hydroxy group on the benzyl substituent and tricyclic thiolactam ring were essential for the cytotoxicity in NDM against PANC-1 cell line. Moreover, 3′-hydroxy derivative 3e compound exhibited antitumor activity against the pancreatic ductal adenocarcinoma (PDAC) xenograft model in vivo.

Curcumin inhibits hypoxia-induced epithelial‑mesenchymal transition in pancreatic cancer cells via suppression of the hedgehog signaling pathway.

Hypoxic microenvironment, a common feature of pancreatic cancer, is associated with tumor proliferation, metastasis and epithelial-mesenchymal transition (EMT) changes. In recent years, many natural agents, including curcumin, have been proven to possess the ability to inhibit the progression of pancreatic cancer. However, whether curcumin is able to suppress hypoxia-induced pancreatic cancer progression and the underlying mechanisms are still not fully elucidated. The aim of the present study was to evaluate whether curcumin affects hypoxia-induced EMT and the activation of Hh signaling pathway in pancreatic cancer. The human pancreatic cancer cell line Panc-1, was treated with hypoxic condition and curcumin. Cell proliferation was assessed by the MTT assay. Wound healing assay and transwell invasion assay were used to detect the migratory and invasive activity of cancer cells. The EMT-related factors, E-cadherin, N-cadherin, vimentin were detected by QT-PCR, western blot analysis and immunofluorescence staining. The Hh signaling-related factors, SHH, SMO and GLI1 were detected by western blot analysis. The results of present study showed that curcumin could not only inhibit the hypoxia-induced cell proliferation, migration and invasion in pancreatic cancer, but also mediate the expression of EMT-related factors. In addition, curcumin remarkably inhibited hypoxia-mediated activation of Hh signaling pathway. Taken together, these data indicate that curcumin plays an important role in suppressing hypoxia-induced pancreatic cancer metastasis by inhibiting the Hh signaling pathway. Curcumin might be a potential candidate for chemoprevention of this severe disease.

Investigating the role of mucin in the delivery of nanoparticles to cellular models of human cancer disease: an in vitro study

Mucin, a glycosylated protein, is aberrantly overexpressed in a variety of tumor cells. The glycoprotein mesh decreases the rate of intracellular drug uptake and effectiveness. We investigated the influence of the mucin mesh on the cellular uptake of anti-MUC1 antibody and nanoparticles by fluorescence spectroscopy and microscopy. A glycosylation inhibitor (benzyl-α-GalNAc) was employed to regulate mucin glycosylation events. In our panel of pancreatic cell lines, only PANC-1 cells exhibited a significant increase in the uptake of liposomes following glycosylation inhibition, resulting in improved cytotoxicity of gemcitabine-loaded liposomes. Interestingly, areas devoid of liposome uptake were observed for pancreatic cancer cell lines PANC-1, Capan-1, and Capan-2; however, these restricted regions could be diminished for PANC1 cells only. In conclusion, investigating the reason(s) for differential cellular uptake of nanoparticles, in association with the production of mucin glycosylation mesh, should provide valuable leads to the future development of nanomedicine for cancer treatment.

Cdc7 is a potent anti-cancer target in pancreatic cancer due to abrogation of the DNA origin activation checkpoint.

PurposeCdc7 is a serine/threonine kinase which is responsible for the ‘firing’ of replication origins leading to initiation of DNA replication. Inhibition or depletion of Cdc7 in normal cells triggers a DNA origin activation checkpoint causing a reversible G1 arrest. Here we investigate Cdc7 as a novel therapeutic target in pancreatic cancer.Experimental designCdc7 target validation was performed by immunoexpression profiling in a cohort of 73 patients with pancreatic adenocarcinoma including 24 controls. Secondly Cdc7 kinase was targeted in Capan-1 and PANC-1 pancreatic cancer cell line models using either an siRNA against Cdc7 or alternatively a small molecule inhibitor (SMI) of Cdc7 (PHA-767491).ResultsCdc7 was significantly overexpressed in pancreatic adenocarcinoma compared to benign pancreatic tissue (median LI 34.3% vs. 1.3%; P<0.0001). Cdc7 knockdown using siRNA in Capan-1 and PANC-1 cells resulted in marked apoptotic cell death when compared with control cells. A prominent sub-G1 peak was seen on flow cytometry (sub-G1 51% vs. 3% and 45% vs. 0.7% in Capan-1 and PANC-1 cells, respectively). Annexin V labelling confirmed apoptosis in 64% vs. 11% and 75% vs. 8%, respectively. Western blotting showed cleavage of PARP-1 and caspase-3 and presence of γH2A.X. TUNEL assay showed strong staining in treated cells. These results were mirrored following Cdc7 kinase inhibition with PHA-767491.ConclusionsOur findings show that Cdc7 is a potent anti-cancer target in pancreatic adenocarcinoma and that Cdc7 immunoexpression levels might be used as a companion diagnostic to predict response to therapeutic siRNAs or SMIs directed against this kinase.

Targeting the insulin growth factor-1 receptor with fluorescent antibodies enables high resolution imaging of human pancreatic cancer in orthotopic mouse models.

The goal of the present study was to determine whether insulin-like growth factor-1 receptor (IGF-1R) antibodies, conjugated with bright fluorophores, could enable visualization of pancreatic cancer in orthotopic nude mouse models. IGF-1R antibody (clone 24-31) was conjugated with 550 nm or 650 nm fluorophores. Western blotting confirmed the expression of IGF-1R in Panc-1, BxPC3, and MIAPaCa-2 human pancreatic cancer cell lines. Labeling with fluorophore-conjugated IGF-1R antibody demonstrated fluorescent foci on the membrane of the pancreatic cancer cells. Subcutaneous Panc-1, BxPC-3, and MIA PaCa-2 tumors became fluorescent after intravenous administration of fluorescent IGF-1R antibodies. Orthotopically-transplanted BxPC-3 tumors became fluorescent with the conjugated IGF-1R antibodies, and were easily visible with intravital imaging. Gross and microscopic ex vivo imaging of resected pancreatic tumor and normal pancreas confirmed that fluorescence indeed came from the membrane of cancer cells, and it was stronger from the tumor than the normal tissue. The present study demonstrates that fluorophore-conjugated IGF-1R antibodies can visualize pancreatic cancer and it can be used with various imaging devices such as endoscopy and laparoscopy for diagnosis and fluorescence-guided surgery.

ALDOA functions as an oncogene in the highly metastatic pancreatic cancer

Pancreatic cancer is an aggressive and devastating disease that is characterized by uncontrolled progression, invasiveness and resistance to conventional treatment. In the past decades, much effort has been given to cancer genetics and pathological classification of this disease. Our previous study has uncovered a subgroup of patients with poor outcome, which is characterized by serum signature of CEA+/CA125+/CA19-9 ≥ 1000 U/mL; however, the underlying biology mechanism remains poorly understood. By using high-throughput screening analysis, we analyzed gene expression signature in highly malignant patients with serum markers of CEA+/CA125+/CA19-9 ≥ 1000 U/mL. Multiple differentially expressed genes were identified, many of which were closely related with cancer metabolic changes. Treatment of pancreatic cancer cell lines PANC-1 with transforming growth factor-β (TGF-β), which was commonly used to induce metastasis, has uncovered that the glycolytic process and antioxidant response was up-regulated upon TGF-β stimulation. These results were consistent with the high-throughput screening analysis. Subsequent analysis indicated that among glycolytic genes, aldolase A (ALDOA) increased the most significantly upon TGF-β treatment. Further in vitro and in vivo results demonstrated that ALDOA was associated with proliferation and metastasis of pancreatic cancer cells. Moreover, ALDOA predicted poor prognosis of pancreatic cancer, partially due to its role in E-cadherin expression regulation, and the results were further validated by analysis of the correlation between ALDOA and E-cadherin expression in pancreatic cancer tissue samples. Mechanistically, the role of ALDOA in pancreatic cancer might attribute to its regulation of c-Myc, HIF1α and NRF2 (Nuclear Factor, Erythroid 2-Like 2), which were key regulators of glycolysis and antioxidant response control.

Interplay Between Gemcitabine and Erlotinib Over Pancreatic Adenocarcinoma Cells.

Pancreatic ductal adenocarcinoma remains as a chemoresistant disease with the poorest prognosis. Gemcitabine has been the standard treatment during the last decade. Erlotinib, a tyrosine kinase inhibitor, in combination with gemcitabine produces a small increase in survival. However, these results remain insufficient. The aim of this study was to investigate the molecular interplay in vitro between them regarding their effects over cytotoxicity, proliferation, apoptosis, and invasion. Using the human pancreatic cancer cell lines Panc-1 and BxPC-3 in vitro, the effects of gemcitabine and erlotinib therapy on growth, proliferation, and invasion were tested by cytotoxicity, cell cycle, and Annexin V-Fluorescein Isothiocyanate analysis, reverse transcription polymerase chain reaction, protein expression, and Chip assays. Therapy decreased cell proliferation causing G0/G1 phase cell cycle arrest with induction of apoptosis in the Panc-1 cell line. This blockade was associated with increased p27 expression. Besides, treatments enhanced the nuclear factor-κB (NF-κB) pathway and the binding of NF-κB to the promoters of genes related to the proliferation and the evasion of apoptosis. Our data suggest that, although gemcitabine and erlotinib exert antiproliferative effects over pancreatic cancer cell lines, the gemcitabine-induced activation of NF-κB expression and its DNA-binding activities are important drawbacks of this treatment against pancreatic cancer.

Up-Regulation of microrna-125b Induced By 5-Fluorouracil (5-FU) Treatment Inhibits Cell Growth and Proliferation in Pancreatic Cancer Cells

MicroRNAs (miRNAs) are a class of small non-coding RNAs that play regulatory roles in protein synthesis by repressing translation or cleaving RNA transcripts. Their aberrant expression has been shown to be involved in human diseases, including cancer. The functional importance of miRNAs is evidenced by many biological processes in which they are implicated, including developmental timing, cell proliferation, apoptosis, metabolism, cell differentiation and morphogenesis. However, little is known about the roles of miRNAs in 5-Fluorouracil (5-FU) resistance and metabolism pathway in pancreatic cancer cells. To address this enigma, miRNA profiles that correlate with 5-FU treatments are defined using miRNA microarray, and comparison of miRNA profiles reveals specific differences between untreated and 5-FU treated pancreatic cancer cells. A subset of miRNAs is being upregulated in 5-FU treated pancreatic cancer cells, with the most significantly upregulated miRNAs being miR-125b, miR-181a, miR-181b, miR-181d, miR-27a, miR-222, miR-30a-5pre and miR-495. Transient transfection of the Pre-miRTM miRNA precursor molecules for miR-125b showed that it inhibited cell proliferation. Expression plasmids of miR-125b were constructed and transfected into human pancreatic cancer cell line PANC-1. PANC-1 miR-125b transfected stable clones showed decreased cell growth and proliferation. Our study suggested that miR-125b may act as a tumor suppressor and present a novel strategy for anticancer therapy.

Ability of simple organotin polyethers to inhibit pancreatic cancer

ABSTRACTA wide variety of organotin polyethers derived from the interfacial polymerization of dibutyltin dichloride and non-cancer inhibitory diols show decent to good inhibition of two human pancreatic cancer cell lines. In view of the general lack of ability of tested compounds to inhibit pancreatic cancer cell lines, this is significant. These cell lines are the AsPC-1 pancreatic cancer cell line which is an adenocarcinoma pancreatic cell line and the PANC-1 pancreatic cancer cell line which is an epithelioid carcinoma pancreatic cell line. The dibutyltin polyethers generally both exhibit EC50 values in the same range and lower than cisplatin and higher CI50 values than cisplatin. Essentially all of the polymers derived from simple non-aromatic diols showed CI50 values greater than 2.

A comparative study of open and laparoscopic radical antegrade modular pancreatosplenectomy versus conventional distal pancreatosplenectomy for left-sided pancreatic ductal adenocarcinoma

Background: Netrin-1 displays proto-oncogenic activity in several cancers, and exerts its biological effects through binding to its receptors.Netrin-1 is up-regulated in several types of cancer tissues and either knockdown of its expression or inhibition of its receptors result in the tumor regression.SENP1 encodes a cysteine protease that specifically targets members of the small ubiquitin-like modifier (SUMO) protein family.SENP1 is involved in the pathogenesis of tumorigenesis and its expression levels are elevated in several types of cancers, we investigate the roles of Netrin-1 on the regulation of SENP1 in pancreatic cancer cells. Methods: In the present study, we determined the role of Netrin-1 on SENP1 expression in the pancreatic cancer cell line PANC-1 and CFPAC-1 cells by RNA isolation, Real-time PCR, Western blot analysis, Small interfering RNA, Dual luciferase reporter assays and Chromatin immunoprecipitation assays. Results: Our results revealed that Netrin-1 overexpression promoted while its knockdown inhibited SENP1 expression in two pancreatic cancer cells. At the molecular level, we found that Netrin-1 activates NF-kB signaling pathway and regulates SENP1 expression through NF-kB signaling pathway. Conclusion: Our studies demonstrate a previous unknown Netrin-1/ SENP1 network in pancreatic cancer cells, suggesting that the abnormal function of Netrin-1 could be an important mechanism for the initiation and progression of pancreatic cancer 15431. Overexpression of nodal enhances perineural invasion in pancreatic cancer by promoting tumor-neural convergence Jiahui Li, Zheng Wang, Zheng Wu, Qingyong Ma Department of Hepatobiliary and Pancreas Surgery, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China Aim: To explore the contribution of Nodal in pancreatic cancer Perineural invasion (PNI) and progression. Background: PNI is a characteristic feature for local spreadof pancreatic cancer. Nodal, an embryonic proteinplays a critical role in embryonic neural development, overexpressed in human pancreatic cancer. Methods: We evaluated the function of Nodal in PNI progression by conducting rat dorsal root ganglia and pancreatic cancer cells co-culture in vitro, cellular and molecular biology, and animal models. Results: We find that overexpression of Nodal in tumor cells contribute to PNI in pancreatic cancer and can serve as a marker for progression of pancreatic cancer. Moreover, the overexpression of Nodal leads to promotion of pancreatic cancer cell migration along nerve axons and upregulating expression of NGF, BDNF and GDNF in tumor cells, which * Corresponding author. enhances nerve outgrowth to pancreatic cancer cell colonies in an in vitro 3D model of nerve invasion in cancer. Furthermore, upregulated Nodal expression enhanced tumor cell invasion of the trunk and nerve dysfunction along sciatic nerves and also promoted orthotropic xenograft tumor growth in vivo. Conclusions:Our study indicates thatNodal is involved in tumor invasion via mediating nerve-tumor interaction. Inhibiting the expression of Nodal could be a potential strategy targeting PNI in pancreatic cancer therapy. 15433. A comparative study of open and laparoscopic radical antegrade modular pancreatosplenectomy versus conventional distal pancreatosplenectomy for left-sided pancreatic ductal adenocarcinoma MenghuaDai, GuangdongWu, Yatong Li, Ning Shi, Taiping Zhang, Quan Liao, Ziwen Liu, Junchao Guo, Wenming Wu, Ge Chen, Ya Hu, Yupei Zhao Department of General Sugery, Peking Union Medical Colleage Hospital, Chinese Academy of Medical Science, Beijing, China, 100730 E-mail address: daim66@126.com, zhao8028@263.net Objective: This study was designed to determine whether Radical antegrade modular pancreatosplenectomy (RAMPS) could improve resection margin status and number of lymph node resected. This study also would prove whether laparoscopic (lap) RAMPS for left-sided pancreatic ductal adenocarcinoma (PDAC) is feasible and safe. Method: Data from 36 patients performed RAMPS were collected from June 2009 to October 2014, 8 of them performed lap RAMPS. 83 patients performed conventional distal pancreatosplenectomy (CDP) for left-side PDAC at the same duration were as case-control to compare clinicopathological factors and oncological outcomes. We also compared these variables between lap and open RAMPS. Result: In RAMPS group, the median number of lymph nodes resected was 14. There was a significant difference in number of lymph nodes resected (14 vs 8; p1⁄40.000) for RAMPS and CDP. On survival analysis, median survival of 24.53 months in RAMPS group was better than that of 21.00months in CDP group (p1⁄40.096). Surgical procedure (RAMPS or CDP), grade of differentiation, TNM stage and resection margin status were independent factors for postoperative prognosis of patients by multivariate analysis (p1⁄40.036, p1⁄40.002, p1⁄40.009, p1⁄40.002).The median length of hospital stay (LOS) and postoperative time to recovering diet in lap RAMPS group were 17 and 3.5 days, which were significant shorter than in the open RAMPS group (19.5 and 6 days), (p1⁄40.045, p1⁄40.004). Conclusion: RAMPS increased the number of lymph node resected and was prognostic factor for improved survival in left-sided PDAC patients. Lap RAMPS was feasible and safe for the left-side PDAC with short LOS and postoperative time to recovering diet.

Polyethylenimine modified calcium phosphate nanoparticles for efficient siRNA delivery and K-ras gene knockdown

Event Abstract Back to Event Polyethylenimine modified calcium phosphate nanoparticles for efficient siRNA delivery and K-ras gene knockdown Xiaochun Xu1*, Wei Yang1*, Chungang Xie1*, Cen Chen1* and Xiangdong Kong1* 1 Zhejiang Sci-Tech University, College of Life Sciences, China Introduction: As one of the important ways of gene therapy, RNA interference has attracted more and more attention. Developing a safe and efficient carrier to guide siRNA into target cytoplasm is a critical challenge in RNA interference field. Calcium phosphate nanoparticles were developed as a carrier nearly 40 years ago and it got enormous attention due to its ability to condense and release cargo efficiently. Materials and Methods: In this study, calcium phosphate nanoparticles were synthesized with the regulation of PEG and then modified by polyethylenimine (PEI). The prepared nanoparticles were mixed with siRNA and then used to transfect human pancreatic cancer cell lines. The efficiency of the delivered siRNA and its K-ras gene knockdown effect were also studied. Results and Discussion: The prepared spherical nanoparticle was confirmed to have a diameter of ＜ 50 nm with a narrow size distribution. As a natural inorganic material, the nanoparticle has little cytotoxicity on normal human pancreatic ductal epithelial cell line HPDE6-C7 and human pancreatic cancer cell lines PANC-1, BXPC-3, CFPAC-1. After 3 months’ degradation in PBS solution with pH value of 5.6, 6.5, 7.4, the degradation percentage of nanoparticles reached 80%, 50%, 40% respectively. When the quality ratio of the nanoparticle and RNA was 8:1, the loading efficiency of RNA was almost 100%. The in vitro test indicated that luciferase FAM-siRNA delivered by the nanoparticle could enter PANC-1 and CFPAC-1 cells successfully, and the transfection efficiency is higher than the conventional siRNA-MateTM . Real-time fluorescent quantitative PCR and western blotting results demonstrated that the siRNA loaded by nanoparticle could silence k-ras gene in RNA and protein expression levels. Meanwhile, knockdown k-ras by transfection complex could significantly induce the apoptosis of PANC-1 cells in vitro. Conclusion: Cytotoxicity studies demonstrated that the prepared nanoparticles have excellent biocompatibility which is most important to be used as an siRNA carrier. Degradation experiment showed the good biodegradation ability of the synthesized nanoparticles. Transfection and silence experiments proved that the carrier can be applied to knockdown k-ras gene and promote the apoptosis of tumor cells successfully. All the results illustrated the prepared nanoparticles could be used as a safe and effective siRNA delivery system for anticancer therapy. National Natural Science Foundation of China (51272236, 51002139); Program for 521 Excellent Talents of Zhejiang Sci-Tech University Keywords: Regenerative Medicine, Calcium phosphate, siRNA delivery, Non-viral vector Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Enabling RNAi with biomaterials Citation: Xu X, Yang W, Xie C, Chen C and Kong X (2016). Polyethylenimine modified calcium phosphate nanoparticles for efficient siRNA delivery and K-ras gene knockdown. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.01249 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. 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Received: 27 Mar 2016; Published Online: 30 Mar 2016. * Correspondence: Dr. Xiaochun Xu, Zhejiang Sci-Tech University, College of Life Sciences, Hangzhou, China, Email1 Dr. Wei Yang, Zhejiang Sci-Tech University, College of Life Sciences, Hangzhou, China, Email2 Dr. Chungang Xie, Zhejiang Sci-Tech University, College of Life Sciences, Hangzhou, China, Email3 Dr. Cen Chen, Zhejiang Sci-Tech University, College of Life Sciences, Hangzhou, China, Email4 Dr. Xiangdong Kong, Zhejiang Sci-Tech University, College of Life Sciences, Hangzhou, China, Email5 Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Xiaochun Xu Wei Yang Chungang Xie Cen Chen Xiangdong Kong Google Xiaochun Xu Wei Yang Chungang Xie Cen Chen Xiangdong Kong Google Scholar Xiaochun Xu Wei Yang Chungang Xie Cen Chen Xiangdong Kong PubMed Xiaochun Xu Wei Yang Chungang Xie Cen Chen Xiangdong Kong Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.