PANC-1 Pancreatic Cancer Cells Research Articles

Gemcitabine‑fucoxanthin combination in human pancreatic cancer cells.

Gemcitabine is a chemotherapeutic agent for pancreatic cancer treatment. It has also been demonstrated to inhibit human pancreatic cancer cell lines, MIA PaCa-2 and PANC-1. The aim of the present study was to investigate the suppressive effect of fucoxanthin, a marine carotenoid, in combination with gemcitabine on pancreatic cancer cells. MTT assays and cell cycle analysis using flow cytometry were performed to study the mechanism of action. The results revealed that combining a low dose of fucoxanthin with gemcitabine enhanced the cell viability of human embryonic kidney cells, 293, while a high dose of fucoxanthin enhanced the inhibitory effect of gemcitabine on the cell viability of this cell line. In addition, the enhanced effect of fucoxanthin on the inhibitory effect of gemcitabine on PANC-1 cells was significant (P<0.01). Fucoxanthin combined with gemcitabine also exerted significant enhancement of the anti-proliferation effect in MIA PaCa-2 cells in a concentration dependent manner (P<0.05), compared with gemcitabine treatment alone. In conclusion, fucoxanthin improved the cytotoxicity of gemcitabine on human pancreatic cancer cells at concentrations that were not cytotoxic to non-cancer cells. Thus, fucoxanthin has the potential to be used as an adjunct in pancreatic cancer treatment.

Effects of LPA receptor-mediated signaling on the modulation of cellular functions of pancreatic cancer cells cultured in fibroblast supernatants under hypoxic conditions.

The tumor microenvironment (TME) consists of various cell types, including fibroblasts. The TME plays a central role in the promotion of tumor progression. In the present study, we investigated whether lysophosphatidic acid (LPA) receptor-mediated signaling regulates cellular functions by the TME of pancreatic cancer PANC-1 cells. To obtain fibroblast 3T3 cell supernatants, 3T3 cells were cultured in 5% charcoal stripped FCS-DMEM for 48h. LPAR2 and LPAR3 expression levels were elevated in PANC-1 cells cultured in 3T3 cell supernatants. While PANC-1 cell motility was decreased by 3T3 cell supernatants, the cell survival to cisplatin (CDDP) of PANC-1 cells was markedly enhanced. Moreover, the cell survival to CDDP of PANC-1 cells cultured in 3T3 cell supernatants was increased by GRI-977,143 (LPA2 agonist) and (2S)-OMPT (LPA3 agonist). Since hypoxia is caused by the restriction of adequate vascular networks to deliver oxygen into solid tumors, PANC-1 cells were cultured in 3T3 cell supernatants at 1% O2 conditions. The cell survival to CDDP of PANC-1 cells cultured in 3T3 cell supernatants at 1% O2 was significantly elevated, correlating with LPAR2 and LPAR3 expressions. These results suggest that LPA signaling via LPA2 and LPA3 is involved in the promotion of malignant properties by the TME in PANC-1 cells.

Design, Synthesis and Biological Evaluation of Novel Catalpol Derivatives as Potential Pancreatic Cancer Inhibitors.

A series of C10-position imidazole-modified catalpol derivatives are specifically designed and synthesized for serving as potential pancreatic cancer inhibitors, which are characterized by 1 H NMR, 13 C NMR and high-resolution mass spectrometry (HRMS). They were evaluated by the 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) test on two human pancreatic cancer cells PANC-1, BxPC-3 and normal pancreatic cell HPDE6-C7, which showed the significant inhibitory effected on the growth of human pancreatic cancer cells of PANC-1 and BxPC-3, especially 91.6% efficacy on BxPC-3, and 73.1% on PANC-1. Simulation studies like molecular docking supported strong binding of vascular endothelial growth factor receptor 2 (VEGFR-2) protein tyrosine kinase (PDB ID: 4AGD), a target of pancreatic cancer. A novel imidazol-modified catalpol compound 3i with strong inhibitory effect on pancreatic cancer cells, which could potentially develop into anti-pancreatic cancer drug candidates in the future.

Design, synthesis and anticancer activities evaluation of novel pyrazole modified catalpol derivatives

Catalpol, a natural product mainly existed in plenty of Chinese traditional medicines, is an iridoid compound with the comprehensive effects on neuroprotective, anti-inflammatory, choleretic, hypoglycemic and anticancer. However, there are some disadvantages for catalpol such as a short half-life in vivo, low druggability, stingy binding efficiency to target proteins and so on. It is necessary to make structural modification and optimization which enhance its performance on disease treatments and clinic applications. Pyrazole compounds have been reported to have excellent anticancer activities. Based on the previous research foundation of our research group on iridoids and the anticancer activities of catalpol and pyrazole, a series of pyrazole modified catalpol compounds were synthesized by principle of drug combination for serving as potential cancer inhibitors. These derivatives are characterized by 1H NMR, 13C NMR and HRMS. The efficacy of anti-esophageal cancer and anti-pancreatic cancer activities were evaluated by the MTT assay on two esophageal cancer cells Eca-109 and EC-9706, and two pancreatic cancer cells PANC-1, BxPC-3 and normal pancreatic cell line HPDE6-C7, which showed that the compound 3e had strong inhibitory activity against esophageal cancer cells, this providing a theoretical basis for the discovery of catalpol-containing drugs.

O011 Investigating the effect of supraphysiological levels of calcium on irreversible electroporation (IRE) of pancreatic cancer cell lines

Abstract Introduction Pancreatic cancer remains difficult to treat with a 5-year survival of 2%. Loco-regional therapies are being introduced such as Irreversible Electroporation (IRE); a non-thermal ablative therapy thought to cause cell death by increasing the cell membrane potential resulting in the formation of aqueous pores, leading to cell death. Mitochondrial changes have previously been reported to be a key driver of apoptosis and calcium has been implicated in this process. We aimed to study the influence of calcium in combination with IRE in pancreatic cancer. Methods Immortalised pancreatic cancer cell line, PANC-1, and pancreatic cancer patient-derived xenograft cells (PDX 185) were exposed to calcium (Ca2+) and IRE monotherapy and in combination. Cells were trypsinised and electroporated in 4mm cuvettes with a BTX generator (Harvard Apparatus). Cell proliferation and viability were analysed, and depolarised mitochondria were evaluated by TMRE and analysed using flow cytometry. Results IRE combined with calcium was more cytotoxic compared to monotherapy with cell proliferation assays demonstrating a statistically significant reduction in proliferation (p&amp;lt;0.0001) across both cell lines. Interestingly, mitochondrial membrane potential analyses demonstrated a depolarising effect only in cells exposed to both calcium and IRE suggesting a mitochondrial mediated effect in cell death in these groups. Conclusion Calcium in combination with IRE may act to potentiate cell death through depolarisation of the mitochondrial membrane. Future work will involve understanding the impact of IRE in combination with calcium in 3D models, allowing the evaluation of molecular cell death mechanisms.

Agrimoniin is a dual inhibitor of AKT and ERK pathways that inhibit pancreatic cancer cell proliferation.

Molecular-targeted therapy has shown its effectiveness in pancreatic cancer, while single-targeted drug often cannot provide long-term benefit because of drug resistance. Fortunately, multitarget combination therapy can reverse drug resistance and achieve better efficacy. The typical treatment characteristics of traditional Chinese medicine monomer on tumor are multiple targets, with small side effects, low toxicity, and so forth. Agrimoniin has been reported to be effective on some cancers, while the mechanism still needs to be clarified. In this study, we used 5-ethynyl-2'-deoxyuridine, cell counting kit-8, flow cytometry, and western blot experiments to confirm that agrimoniin can significantly inhibit the proliferation of pancreatic cancer cell PANC-1 by inducing apoptosis and cell cycle arrest. In addition, by using SC79, LY294002 (the agonist or inhibitor of AKT pathway), and U0126 (the inhibitor of ERK pathway), we found that agrimoniin inhibited cell proliferation by simultaneously inhibiting AKT and ERK pathways. Moreover, agrimoniin could significantly increase the inhibitory effect of LY294002 and U0126 on pancreatic cancer cells. Meanwhile, in vivo experiments also supported the above results. In general, agrimoniin is a double-target inhibitor of AKT and ERK pathways in pancreatic cancer cells; it is expected to be used as a resistance reversal agent of targeted drugs or a synergistic drug of the inhibitor of AKT pathway or ERK pathway.

ZEB2 Regulates the Migration and Invasion of PANC-1 Pancreatic Cancer Cells: An Experimental Study

To investigate the effects and mechanisms of zinc finger E-box binding homeobox transcription factor-2 ( ZEB2) on the proliferation, colony formation, migration, and invasion abilities and the epithelial-mesenchymal transition (EMT) of PANC-1 cells, a human pancreatic cancer cell line. Data on the expression of ZEB2 in pancreatic cancer tissues and paracancerous tissues from The Cancer Genome Atlas (TCGA) database were analyzed. PANC-1 pancreatic cancer cells were divided into si-NC group, si- ZEB2 group, pcDNA3.1 group, and pcDNA3.1- ZEB2 group. qRT-PCR and Western blot were conducted to confirm the effectiveness of ZEB2 knockdown or overexpression. CCK-8, colony formation, wound healing, and Transwell assays were conducted to examine the effects of ZEB2 on the proliferation, colony formation, migration, and invasion of PANC-1 cells. qRT-PCR and immunofluorescence assays were performed to examine the expression of E-cadherin and vimentin, the EMT markers, in the cells. Prediction of proteins interacting with ZEB2 was made through the STRING database. TCGA database analysis showed that the expression level of ZEB2 in pancreatic cancer tissues was significantly higher than that in adjacent tissues ( P<0.05). Compared with those of cells in the control group, the proliferation, colony formation, migration, and invasion of cells in the si- ZEB2 group were decreased ( P<0.05). Compared with those of cells in the pcDNA3.1 group, the proliferation, colony formation, migration and invasion of cells in the pcDNA3.1- ZEB2 group were increased (all P<0.05). According to the results of qRT-PCR and immunofluorescence assays, compared with those of the si-NC group, the expression of E-cadherin mRNA, an epithelial marker, in the si- ZEB2 group increased, while the expression of vimentin mRNA, an mesenchymal marker, and the protein decreased. Compared with those of the pcDNA3.1 group, the expression of E-cadherin mRNA in the PANC-1 cells of the pcDNA3.1- ZEB2 group decreased, while the expression of vimentin mRNA and the protein increased (all P<0.05). Analysis with the STRING database predicted that 10 proteins had close interaction with ZEB2. Overexpression of ZEB2 promotes the migration, invasion, and the EMT process of PANC-1 pancreatic cancer cells.

Xiao Chai Hu Tang alleviates the pancreatic tumorigenesis via improving the mtDNA N6-Methyladenine modification mediated mitochondrial dysfunction in Syrian hamster model

BackgroundPancreatic intraepithelial neoplasia (PanIN) is the most common precursor lesion of pancreatic ductal adenocarcinoma (PDAC), which is a highly malignant tumor and lack of effective treatment. Although Xiao Chai Hu Tang (XCHT) has a good therapeutic effect on pancreatic cancer patients with advanced stage, the effect and mechanism of XCHT remains unclear in pancreatic tumorigenesis. PurposeTo assess the therapeutic effects of XCHT on the malignant transformation from PanIN to PDAC and to reveal its mechanisms of pancreatic tumorigenesis. MethodsSyrian golden hamster were induced by N-Nitrosobis (2-oxopropyl) amine (BOP) to establish the pancreatic tumorigenesis model. The morphological changes of pancreatic tissue were observed by H&E and Masson staining; the Gene ontology (GO) analysis the transcriptional profiling changes; the mitochondrial ATP generation, mitochondrial redox status, mitochondrial DNA (mtDNA) N6-methyladenine (6mA) level and relative mtDNA genes expressions were examined. In addition, immunofluorescence detect the cell localization of 6mA in human pancreatic cancer PANC1 cell. Using the TCGA database, the prognostic effect of mtDNA 6mA demethylation ALKBH1 expression on pancreatic cancer patients was analyzed. ResultsWe confirmed the mtDNA 6mA levels were gradually increased with the mitochondrial dysfunction in PanINs progression. XCHT showed the effect to inhibit the occurrence and development of pancreatic cancer in Syrian hamster pancreatic tumorigenesis model. In addition, the lack of ALKBH1 mediated mtDNA 6mA increase, mtDNA coded genes down-expression and abnormal redox status were rescued by XCHT. ConclusionsALKBH1/mtDNA 6mA mediated mitochondrial dysfunction to induce the occurrence and progression of pancreatic cancer. XCHT can improve ALKBH1 expression and mtDNA 6mA level, regulate the oxidative stress and expression of mtDNA coded genes. This study investigated a new molecular mechanism of pancreatic tumorigenesis, and revealed the therapeutic efficacy of XCHT in pancreatic tumorigenesis for the first time.

LPA receptor-mediated signaling regulates cell motility and survival to anticancer drug of pancreatic cancer cells under glucose-deprived and hypoxic conditions

In tumor microenvironment, cancer cells can adapt to low conditions of nutrients and oxygen. Lysophosphatidic acid (LPA) receptor-mediated signaling is involved in the promotion of malignant properties in cancer cells. In the present study, to examine the roles of LPA receptors in the regulation of cell motility and survival to cisplatin (CDDP) of pancreatic cancer PANC-1 cells under glucose-deprived and hypoxic conditions, cells were cultured in 4500 mg/L high glucose (HG)-DMEM, 500 mg/L middle glucose (MG)-DMEM and 100 mg/L low glucose (LG)-DMEM at 21% and 1% O2. The expression levels of LPAR1 and LPAR2 genes in cells cultured in MG-DMEM and LG-DMEM were significantly elevated, compared with HG-DMEM cells. The cell motility and survival rate to CDDP of cells cultured in MG-DMEM and LG-DMEM were significantly lower than those of cells cultured in HG-DMEM. The cell survival to CDDP was enhanced by LPA1 knockdown and suppressed by LPA2 knockdown. Under hypoxic conditions (1% O2), LPAR1, LPAR2 and LPAR3 expressions were markedly higher in cells cultured in MG-DMEM and LG-DMEM than in cells cultured in HG-DMEM. The cell survival rates to CDDP of cells cultured in MG-DMEM and LG-DMEM were elevated in comparison with HG-DMEM. The cell survival to CDDP was reduced by LPA3 knockdown. These results suggest that LPA receptor-mediated signaling is involved in the regulation of malignant properties of PANC-1 cells under glucose-deprived and hypoxic conditions.

Spatiotemporal Heterogeneity of De Novo Lipogenesis in Fixed and Living Single Cells.

De novo lipogenesis (DNL) is a critical metabolic process that provides the majority of lipids for adipocyte and liver tissue. In cancer, obesity, type II diabetes, and nonalcoholic fatty liver disease DNL becomes dysregulated. A deeper understanding of the rates and of subcellular organization of DNL is necessary for identifying how this dysregulation occurs and varies across individuals and diseases. However, DNL is difficult to study inside the cell because labeling lipids and their precursors is not trivial. Existing techniques either can only measure parts of DNL, like glucose uptake, or do not provide spatiotemporal resolution. Here, we track DNL in space and time as isotopically labeled glucose is converted to lipids in adipocytes using optical photothermal infrared microscopy (OPTIR). OPTIR provides submicron resolution infrared imaging of the glucose metabolism in both living and fixed cells while also reporting on the identity of lipids and other biomolecules. We show significant incorporation of the labeled carbons into triglycerides in lipid droplets over the course of 72 h. Live cells had better preservation of lipid droplet morphology, but both showed similar DNL rates. Rates of DNL, as measured by the ratio of 13C-labeled lipid to 12C-labeled lipid, were heterogeneous, with differences within and between lipid droplets and from cell to cell. The high rates of DNL measured in adipocyte cells match upregulated rates of DNL previously reported in PANC1 pancreatic cancer cells. Taken together, our findings support a model where DNL is locally regulated to meet energy needs within cells.

LncRNA-NEAT1 inhibits the occurrence and development of pancreatic cancer through spongy miR-146b-5p/traf6

ABSTRACT To investigate the inhibitory effect of LNCNA-NEA1 on pancreatic cancer development and progression via spongiosa miR-146b-5p/TRAF6, 60 pancreatic cancer patients diagnosed from December 2017 to December 2019 were selected as a general source of information. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression level of NEAT1 in cancerous and adjacent non-cancerous tissues. Cell counting kit-8 (CCK-8) and transwell were used to determine the effect of LNCNA-NEA1 on the proliferation and migration of pancreatic cancer cells (Panc-1). The results of dual luciferase reporter gene assay showed that nea 1 could target and regulate the expression of spongy miR-146b-5p/TRAF6, and reducing the expression of spongy miR-146b-5p/TRAF6 could reverse the inhibitory effects of nea 1-siRNA on proliferation, migration and invasion of pancreatic cancer cells. Therefore, it was concluded that knockdown of nea 1 could inhibit the proliferation, migration and invasion of pancreatic cancer cells by upregulating the level of miR-146b-5p/TRAF6, and the expression of lnc RNA-nea 1 could be used as an indicator for preoperative diagnosis and postoperative prognosis of pancreatic cancer patients.

Library Screening and Preliminary Characterization of Synthetic Cannabinoids Against Prostate and Pancreatic Cancer Cell Lines.

Background: Our previous screening efforts with colorectal cancer cell lines suggested potential cannabinoid therapeutic leads for other solid cancers. Objectives: The aim of this study was to identify cannabinoid lead compounds that have cytostatic and cytocidal activities against prostate and pancreatic cancer cell lines and profile cellular responses and molecular pathways of select leads. Materials and Methods: A library of 369 synthetic cannabinoids was screened against 4 prostate and 2 pancreatic cancer cell lines with 48 h of exposure at 10 μM in medium with 10% fetal bovine serum using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) viability assay. Concentration titration of the top 6 hits was carried out to identify their concentration-response patterns and calculate IC50 values. Three select leads were examined for cell cycle, apoptosis, and autophagy responses. The role of cannabinoid receptors (CB1 and CB2) and noncanonical receptors in apoptosis signaling was examined with selective antagonists. Results: Two independent screening experiments in each cell line detected growth inhibitory activities against all six or a majority of cancer cell lines for HU-331 (a known cannabinoid topoisomerase II inhibitor), (±)5-epi-CP55,940, and PTI-2, each previously identified in our colorectal cancer study. 5-Fluoro NPB-22, FUB-NPB-22, and LY2183240 were novel hits. Morphologically and biochemically, (±)5-epi-CP55,940 elicited caspase-mediated apoptosis of PC-3-luc2 (a PC-3 subline with luciferase) prostate cancer and Panc-1 pancreatic cancer cell lines, each the most aggressive of the respective organ site. The apoptosis induced by (±)5-epi-CP55,940 was abolished by the CB2 antagonist, SR144528, but not modulated by the CB1 antagonist, rimonabant, and GPR55 antagonist, ML-193, nor TRPV1 antagonist, SB-705498. In contrast, 5-fluoro NPB-22 and FUB-NPB-22 did not cause substantial apoptosis in either cell line, but resulted in cytosolic vacuoles and increased LC3-II formation (suggestive of autophagy) and S and G2/M cell cycle arrests. Combining each fluoro compound with an autophagy inhibitor, hydroxychloroquine, enhanced the apoptosis. Conclusions: 5-Fluoro NPB-22, FUB-NPB-22, and LY2183240 represent new leads against prostate and pancreatic cancer cells in addition to the previously reported compounds, HU-331, (±)5-epi-CP55,940, and PTI-2. Mechanistically, the two fluoro compounds and (±)5-epi-CP55,940 differed regarding their structures, CB receptor involvement, and death/fate responses and signaling. Safety and antitumor efficacy studies in animal models are warranted to guide further R&D.

İnsan Pankreas Kanserinde Borik Asit Uygulaması ile Bazı IncRNA’ların ve İnvazyonun Baskılanması

Objective: It was aimed to investigate the effects of boric acid on some lncRNAs and invasion in PANC-1 and MIA PaCa-2 pancreatic cancer cells. Materials and Methods: The effects of boric acid on cell viability and invasion were investigated using XTT test and invasion chambers, respectively. lncRNA H19 and UCA1 expressions were evaluated in pancreatic cancer using TCGA data. Its effects on expressions of these lncRNAs and invasion genes were determined by qRT-PCR analysis. Results: The IC50 doses of boric acid were calculated as 14.25 mM in PANC-1 cells and 15.71 mM in MIA PaCa-2. TCGA data showed that H19 and UCA1 expressions were elevated in pancreatic cancer. H19 and UCA1 lncRNA levels were decreased with boric acid treatment. In addition, boric acid increased CDH1 and TIMP1 in both cell lines. However, it suppressed CDH2 expression. Boric acid increased TIMP2 in PANC-1 cells and TIMP3 expression in MIA PaCa-2 cells. In the invasion test, boric acid significantly suppressed invasion in both cells. Conclusions: Boric acid suppressed H19 and UCA1 expressions, which were found to be high in pancreatic cancer. In addition, it showed an anti-invasive effect by changing the expressions of genes that are important in invasion.

Preparation and Characterization of Rutin-Encapsulated Polymeric Micelles and Studies of Synergism with Bioactive Benzoic Acids and Triazolofluoroquinolones as Anticancer Nanomedicines.

The study aimed to examine rutin micelles of advanced superlative dual cytotoxicity-antiinflammtion bioefficacies in substantially novel submicro-nanoaffinities vs. both the raw rutin and reference proapoptotic cisplatin. Antiproliferative capabilities of rutin, benzoic acid (BA) and triazolofluoroqunolone (TFQ) derivatives were reported; hence chemosensitizing effects of rutin or its polymeric micelles (of improved solubility and bioavailability via direct dissolution using the amphiphilic copolymer Pluronic P123) in co-incubations with 5 BAs or 3 TFQ derivatives in a panel of 6 cancer cell lines were verified. Rutin loading in micelles was achieved with a loading efficiency of 59.5 ± 2.9%. The particle size of the micelles was found to be 18 ± 2 nm. Though Rutin loaded nanomicelles were of minimal DPPH radical scavenging activity; they had nitrogen oxide (NO) radical scavenging activity in lipopolysaccharide-induced RAW264.7 macrophages with equipotency to indomethacin (IC50 values (µM) 73.03 vs. 60.88; p=0.057). Remarkably nano-micelle formulation of rutin was proved of significantly more potent antineoplastic bioactivity with submicro-nanomolar affinities in the 6 cancer cell lines vs. both free rutin's and cisplatin's (except A549 lung cancer cell line). Rrutin nanomicelles chemo-sensitized all selected 8 cotreatments with BA derivatives and TFQs and, thus reducing the dose used against breast cancer MCF7 cells to submicro-nanomolar affinities of greater potencies than cisplatin's. Except for Triazolo-4-anisidine cipro butyl acid in PANC1, 2-Amino-3,5-Di iodo BA in A375 and 4-Nitrophenol in A549 incubations; rutin loaded nanomicelles chemosensitized 7/8 cotreating selected benzoic acid (BAs) derivatives and TFQs and chemosensitized pancreatic PANC1, skin A375 and lung A549 cancer cell lines, thus reducing the dose to submicro-nanomolar affinities of greater potencies than cisplatin's. Rutin loaded nanomicelles chemosensitize 6/8 cotreating selected benzoic acid (BA) derivatives and TFQs (except for 2-Amino-5-Bromo Benzoic Acid and Triazolo-4-anisidine cipro butyl acid), thus reducing the dose used against resistant CACO2 colorectal cancer cells.

Boric Acid Shows ER Stress and Apoptosis Mediated Anticancer Activity in Human Pancreatic Cancer MIA PaCa-2 and PANC-1 Cells

Amaç: Bu çalışmada borik asitin insan pankreas kanseri MIA PaCa-2 ve PANC-1 hücrelerinde endoplazmik retikulum (ER) stresi ve apoptoz aracılı antikanser etkisinin araştırılması amaçlanmıştır. Gereçler ve Yöntem: Borik asitin pankreas kanseri hücrelerinin canlılığı üzerine etkisi ve IC50 değeri XTT testi ve CompuSyn version 1.0 yazılımı kullanılarak hesaplanmıştır. Apoptotik, anti-apoptotik ve ER stresi ile ilişkili genlerin ifadesi belirlenmiştir. Borik asitin bu hücrelerin koloni oluşum kapasitesi üzerine etkisi ise koloni oluşum testi ile değerlendirilmiştir. Bulgular: Borik asit zaman ve doz bağımlı olarak her iki hücre hattında da hücre canlılığını baskılamıştır. XTT testi sonucunda MIA PaCa-2 ve PANC-1 hücrelerinde borik asitin IC50 dozlarının sırasıyla 15707,5 ve 14248,8 μM olduğu bulunmuştur. Borik asitin her iki hücre hattında da apoptoz ile ilişkili genlerden BAX, CASP3, CASP8, CYCS ve FAS genlerinin ifadelerini anlamlı derecede arttırdığı gözlenmiştir. PANC- 1 hücrelerinde CASP9 ve FADD genlerinin ifadeleride anlamlı derecede yükselmiştir. Borik asitin her iki hücre hattında da ER stresi ile ilişkili ATF4, HSP47 ve XBP1 genlerinin ifadesini istatistiksel olarak anlamlı derecede arttırdığı görülmüştür. Ayrıca borik asit muamelesi sadece PANC-1 hücrelerinde ATF6, CHOP ve EIF2A ifadelerini anlamlı derecede arttırmıştır. MIA PaCa-2 hücrelerinde ise borik asit GRP78 gen ifadesinin istatistiksel olarak artmasına neden olmuştur. Koloni oluşum testi sonuçları borik asitin her iki hücre hattında da koloni oluşum kapasitelerinin anlamlı derecede baskılandığını göstermiştir. Sonuç: Borik asit her iki insan pankreas kanseri hücrelerinde hücre canlılığı ve koloni oluşumunu azaltmış olup apoptoz ve ER stresi ile ilişkili genlerin ifadesini değiştirmiştir. Bu bulgular, borik asitin insan pankreas kanseri hücrelerinde ER stresi ve apoptoz aracılı antikanser etkisini göstermektedir.

Some Chemical Studies on Cytotoxicity of Antioxidants from Propolis Extracts against Human Pancreatic PANC-1 Cancer Cell Line

Some Chemical Studies on Cytotoxicity of Antioxidants from Propolis Extracts against Human Pancreatic PANC-1 Cancer Cell Line

Heptadecanoic Acid, an Odd-Chain Fatty Acid, Induces Apoptosis and Enhances Gemcitabine Chemosensitivity in Pancreatic Cancer Cells.

Odd-chain saturated fatty acids generally serve as specific biomarkers of dietary components and dairy intake, some of which have anticancer properties. This study was performed to assess the anticancer effects of heptadecanoic acid (HDNA) in human pancreatic carcinoma cells. MTT (thiazolyl blue tetrazolium bromide) assay showed that HDNA exerted stronger cytotoxic effects than pentadecanoic acid, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), and linoleic acid (18:2) on both Panc-1 and MIA PaCa-2 pancreatic cancer cells. In addition, HDNA reduced colony formation and induced apoptosis in these pancreatic cancer cells as indicated by Hoechst 33342 staining, Annexin V/propidium iodide staining, cell cycle analysis, and Western blotting analysis in a dose-dependent manner. Moreover, HDNA synergistically reduced cell viability and promoted apoptosis when combined with gemcitabine (GEM), a chemotherapeutic agent commonly used in the treatment of pancreatic cancer. GEM-resistant MIA PaCa-2 (GR-MIA PaCa-2) cells with a resistance indices (RI) value of 215.09 [RI = half-maximal inhibitory concentration (IC50) of GR-MIA PaCa-2 cells/IC50 of MIA PaCa-2 cells] were established, and the efficacy of HDNA on GEM chemosensitivity was confirmed. Surprisingly, HDNA exhibited even higher antiproliferative efficacy against GR-MIA PaCa-2 cells (IC50 = 71.45 ± 6.37 μM) than parental MIA PaCa-2 cells (IC50 = 77.47 ± 2.10 μM). Finally, HDNA treatment inhibited the Hippo pathway and induced apoptosis of GR-MIA PaCa-2 cells. These findings suggest the beneficial effects of a HDNA-rich diet during pancreatic cancer treatments.

The inhibition of pancreatic cancer progression by PI3K-activated MSC proteomes

Background and Hypothesis:Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with a very poor survival rate. The PI3K pathway has been studied extensively in cancer cells which has led to the creation of PI3K inhibitor drugs. To explore an uncharted and counterintuitive use of the PI3K pathway, PI3K was activated in mesenchymal stem cells (MSCs) and the action of their conditioned medium (CM) on PDAC was examined. We hypothesized that PI3K-activated MSC-derived CM is capable of suppressing tumor growth and migration.&#x0D; Project Methods:To generate the treatment conditioned media, we first isolated and grew MSCs in DMEM + 10% serum and added 50 μM of PI3K activator (YS49) and incubated for 24 hours. Afterwards, we exchanged the media with fresh DMEM + 10% serum and incubated for another 24 hours. Next, we hyper-centrifuged the media to extract the conditioned media and used the media to treat PANC-1, PA03C, ASPC-1, PANC10.05, and PANC198 pancreatic cancer cell lines. In vitro viability and migration assays were conducted on these five treated cell lines. The regulatory mechanism was evaluated using Western blotting, immunoprecipitation, and ELISAs.&#x0D; Results:PI3K-activated MSC-derived CM reduced the proliferation (N=5) and migration (N=2) of PDAC (p &lt; 0.01). MSC CM was enriched with polyubiquitin C (Ubc), which acted as an extracellular tumor-suppressing protein by interacting with CXCR4. While the anti-tumor efficacy of Ubc differed depending on the CXCR4 expression level of PDAC, MSC CM was commonly effective in the tested cell lines and tissues (N=5).&#x0D; Conclusion and potential impact:Collectively, this study demonstrated that MSC CM can be converted into a tumor-suppressing agent by the activation of PI3K, and a Ubc-CXCR4 regulatory axis is at least in part responsible. The use of MSC CM treatment can potentially serve as an alternative or supplemental treatment to current chemotherapeutic agents in treating pancreatic cancer without inducing life-threatening side effects.

Exosomes secreted from human umbilical cord mesenchymal stem cells promote pancreatic ductal adenocarcinoma growth by transferring miRNAs

Objective: To observe the effects of exosomes derived from human umbilical cord mesenchymal stem cells on the proliferation and invasion of pancreatic cancer cells, and to analyze the contents of exosomes and explore the mechanisms affecting pancreatic cancer cells. Methods: Exosomes extracted from human umbilical cord mesenchymal stem cells were added to pancreatic cancer cells BxPC3, Panc-1 and mouse models of pancreatic cancer, respectively. The proliferative activity and invasion abilities of BxPC3 and Panc-1 cells were measured by cell counting kit-8 (CCK-8) and Transwell assays. The expressions of miRNAs in exosomes were detected by high-throughput sequencing. GO and KEGG were used to analyze the related functions and the main metabolic pathways of target genes with high expressions of miRNAs. Results: The results of CCK-8 cell proliferation assay showed that the absorbance of BxPC3 and Panc-1 cells in the hucMSCs-exo group was significantly higher than that in the control group [(4.68±0.09) vs. (3.68±0.01), P<0.05; (5.20±0.20) vs. (3.45±0.17), P<0.05]. Transwell test results showed that the number of invasion cells of BxPC3 and Panc-1 in hucMSCs-exo group was significantly higher than that in the control group (129.40±6.02) vs. (89.40±4.39), P<0.05; (134.40±7.02) vs. (97.00±6.08), P<0.05. In vivo experimental results showed that the tumor volume and weight in the exosomes derived from human umbilical cord mesenchymal stem cells (hucMSCs-exo) group were significantly greater than that in the control group [(884.57±59.70) mm(3) vs. (695.09±57.81) mm(3), P<0.05; (0.94±0.21) g vs. (0.60±0.13) g, P<0.05]. High-throughput sequencing results showed that miR-148a-3p, miR-100-5p, miR-143-3p, miR-21-5p and miR-92a-3p were highly expressed. GO and KEGG analysis showed that the target genes of these miRNAs were mainly involved in the regulation of glucosaldehylation, and the main metabolic pathways were ascorbic acid and aldehyde acid metabolism, which were closely related to the development of pancreatic cancer. Conclusion: Exosomes derived from human umbilical cord mesenchymal stem cells can promote the growth of pancreatic cancer cells and the mechanism is related to miRNAs that are highly expressed in exosomes.

Effects of Casein Kinase 2 Alpha Subunit on the Proliferation, Invasion and Migration of Pancreatic Cancer PANC-1 Cells via the PI3K/Akt/GSK-3β Pathway

The researchers aimed to evaluate the functions of casein kinase 2 alpha subunit (CK2a) expression silencing in influencing pancreatic cancer (PC) PANC-1 cells from the aspects of invasion, proliferation and migration. As PC-1, PC PANC-1 and SW1990 cells exhibited significantly reduced CK2a mRNA expression level in contrast to normal pancreatic cells HPDE6-C7, while PC tissues displayed raised expression level compared with adjacent tissues (P&lt;0.05). CK2a mRNA expression had significant correlations with nerve infiltration, lymph node metastasis, tumor differentiation degree and tumor-node-metastasis stage. After interfering with the expression of CK2a in PANC-1 cells, p-PTEN had elevated expression, while p-Akt473, p-GSK-3β, p-c-Raf, p-Akt308, p-PDK1, Snail and Vimentin, βcatenin had significantly down-regulated expressions, impeding cell growth, decreasing the number of healing wounds and invading cells. Moreover, the transplanted tumors were inhibited in terms of growth. For PC PANC-1 cells, their metastasis, proliferation and invasion were modulated by CK2a probably through regulating the PI3K/Akt/GSK-3β pathway that targeted epithelial-mesenchymal transition.