agar and Pancreatic-Neoplasms

Pancreatic cancer is a pathology with a high mortality rate since it is detected at advanced stages, so the search for early-stage diagnostic biomarkers is essential. Liquid biopsies are currently being explored for this purpose and educated platelets are a good candidate, since they are known to present a bidirectional interaction with tumor cells. In this work, we analyzed the effects of platelets on cancer cells' viability, as determined by MTT, migration using transwell assays, clonogenicity in soft agar and stemness by dilution assays and stem markers' expression. We found that the co-culture of platelets and pancreatic cancer cells increased the proliferation and migration capacity of BXCP3 cells, augmented clonogenicity and induced higher levels of Nanog, Sox2 and Oct4 expression. As platelets can provide horizontal transfer of microRNAs, we also determined the differential expression of miRNAs in platelets obtained from a small cohort of pancreatic cancer patients and healthy subjects. We found clear differences in the expression of several miRNAs between platelets of patients with cancer healthy subjects. Moreover, when we analyzed microRNAs from the platelets of the pancreatic juice and blood derived from each of the cancer patients, interestingly we find differences between the blood- and pancreatic juice-derived platelets suggesting the presence of different subpopulations of platelets in cancer patients, which warrant further analysis.

Topics: Agar; Blood Platelets; Cell Line, Tumor; Humans; MicroRNAs; Neoplastic Stem Cells; Pancreatic Neoplasms

We report a case of canine adenocarcinoma with multi-organ metastasis in which colonies of adenocarcinoma cells grew upon aerobic bacterial culture of pleural effusion. Stained agar colonies were highly similar to rare suspicious cells seen on cytologic examination of the pleural effusion, as well as rare cells seen on cytologic examination of pancreatic and gastric wall fine-needle aspirates. Cells from colonies growing on agar media were mildly immunoreactive for cytokeratin. Histologic examination of tissues obtained at autopsy revealed pancreatic adenocarcinoma with vascular invasion and nodal, gastric, pulmonary, and pleural metastasis.

Topics: Adenocarcinoma; Agar; Animals; Bacteriological Techniques; Biopsy, Fine-Needle; Culture Media; Dog Diseases; Dogs; Female; Lung; Lung Neoplasms; Lymphatic Metastasis; Pancreatic Neoplasms; Pleural Effusion, Malignant; Pleural Neoplasms; Stomach Neoplasms

Increased levels of NF-κB are hallmarks of pancreatic ductal adenocarcinoma (PDAC) and both classical and alternative NF-κB activation pathways have been implicated.. Here we show that activation of the alternative pathway is a source for the high basal NF-κB activity in PDAC cell lines. Increased activity of the p52/RelB NF-κB complex is mediated through stabilization and activation of NF-κB-inducing kinase (NIK). We identify proteasomal downregulation of TNF receptor-associated factor 2 (TRAF2) as a mechanism by which levels of active NIK are increased in PDAC cell lines. Such upregulation of NIK expression and activity levels relays to increased proliferation and anchorage-independent growth, but not migration or survival of PDAC cells.. Rapid growth is one characteristic of pancreatic cancer. Our data indicates that the TRAF2/NIK/NF-κB2 pathway regulates PDAC cell tumorigenicity and could be a valuable target for therapy of this cancer.

Topics: Agar; Carcinoma, Pancreatic Ductal; Cell Line, Tumor; Cell Movement; Cell Nucleus; Cell Proliferation; Chemotaxis; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; NF-kappa B p52 Subunit; NF-kappaB-Inducing Kinase; Pancreatic Neoplasms; Proteasome Endopeptidase Complex; Protein Binding; Protein Serine-Threonine Kinases; Tetrazolium Salts; Thiazoles; TNF Receptor-Associated Factor 2

NAD(P)H:quinone oxidoreductase (NQO1) functions as an important part of cellular antioxidant defense by detoxifying quinones, thus preventing the formation of reactive oxygen species (ROS). The aim of our study was to determine if NQO1 is elevated in pancreatic cancer specimens and pancreatic cancer cell lines and if so, would compounds previously demonstrated to redox cycle with NQO1 be effective in killing pancreatic cancer cells. Immunohistochemistry of resected pancreatic specimens demonstrated an increased immunoreactivity for NQO1 in pancreatic cancer and pancreatic intraepithelial neoplasia (PanIN) specimens versus normal human pancreas. Immunocytochemistry and Western immunoblots demonstrated increased immunoreactivity in pancreatic cancer cells when compared to a near normal immortalized human pancreatic ductal epithelial cell line and a colonic epithelial cell line. Streptonigrin, a compound known to cause redox cycling in the presence of NQO1, decreased clonogenic survival and decreased anchorage-independent growth in soft agar. Streptonigrin had little effect on cell lines with absent or reduced levels of NQO1. The effects of streptonigrin were reversed in pancreatic cancer cells pretreated with dicumarol, a known inhibitor of NQO1. NQO1 may be a therapeutic target in pancreatic cancer where survival is measured in months.

Topics: Agar; Blotting, Western; Cell Proliferation; Clone Cells; Humans; Immunohistochemistry; NAD(P)H Dehydrogenase (Quinone); Pancreatic Neoplasms; Phenotype; Streptonigrin; Tissue Array Analysis; Tumor Cells, Cultured

The HER2/neu oncogene is overexpressed in up to 70% of human pancreatic cancer specimens when compared to normal pancreatic tissue. This cell surface receptor can be targeted specifically by the neutralizing antibody Herceptin. Herceptin has been successfully used in combination with other chemotherapeutic agents in breast cancer, a cancer in which only 30% of patients harbor elevated HER2/neu levels. In the present study, we investigated the therapeutic efficacy of Herceptin in combination with gemcitabine and docetaxel. Gemcitabine is currently the standard chemotherapeutic agent used to treat pancreatic cancer. In contrast, docetaxel, a taxane, is only just being investigated in pancreatic cancer. Tumor cell resistance to taxanes is at least in part mediated by the HER2/NEU oncogene. We have previously characterized HER2/NEU expression in human pancreatic cancer cell lines and studied the anti-tumor activity of Herceptin monotherapy in vitro and in vivo. In the present study, combination therapy resulted in a dramatic improvement of animals bearing human pancreatic cancer xenografts. Furthermore, metastasis and production of ascites was lower when a combination of these three agents was used. We conclude that, as with breast cancer, the anti-tumor activity of Herceptin may be improved by combination with taxanes or gemcitabine.

Topics: Agar; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Ascites; Cell Line, Tumor; Cell Survival; Combined Modality Therapy; Deoxycytidine; Docetaxel; Drug Resistance, Neoplasm; Gemcitabine; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neoplasms, Experimental; Pancreatic Neoplasms; Recombinant Proteins; Taxoids; Time Factors; Trastuzumab

Manganese superoxide dismutase (MnSOD) levels have been found to be low in human pancreatic cancer [Pancreas 26, (2003), 23] and human pancreatic cancer cell lines [Cancer Res. 63, (2003), 1297] when compared to normal human pancreas. We hypothesized that stable overexpression of pancreatic cancer cells with MnSOD cDNA would alter the malignant phenotype. MIA PaCa-2 cells were stably transfected with a pcDNA3 plasmid containing sense human MnSOD cDNA or containing no MnSOD insert by using the lipofectAMINE method. G418-resistant colonies were isolated, grown and maintained. Overexpression of MnSOD was confirmed in two selected clones with a 2-4-fold increase in MnSOD immunoreactive protein. Compared with the parental and neo control cells, the MnSOD-overexpressing clones had decreased growth rates, growth in soft agar and plating efficiency in vitro, while in vivo, the MnSOD-overexpressing clones had slower growth in nude mice. These results suggest that MnSOD may be a tumor suppressor gene in human pancreatic cancer.

Topics: Agar; Aged; Animals; Antioxidants; Cell Proliferation; Cells, Cultured; Dicumarol; DNA, Complementary; Gene Expression; Humans; Male; Mice; Mice, Nude; Neoplasm Transplantation; Pancreatic Neoplasms; Superoxide Dismutase; Superoxides; Transfection

Cells contain a large number of antioxidants to prevent or repair the damage caused by reactive oxygen species. One component of the antioxidant system, manganese superoxide dismutase (MnSOD), is localized in the mitochondria, and the levels of this protein have been previously shown to inversely correlate with pancreatic cancer cell growth. The aim of the present study was to determine whether MnSOD overexpression could suppress the in vitro and in vivo malignant phenotype of a human pancreatic cancer cell line. Tumor cell behavior was determined in the pancreatic cancer cell line MIA PaCa-2 by examining cell growth, plating efficiency, and anchorage-independent growth in soft agar. MnSOD was overexpressed in the pancreatic cancer cell line MIA PaCa-2 by infection with an adenovirus-MnSOD construct. Cells were also injected s.c. in nude mice and tumor volume was calculated. Single and multiple direct injections of the adenoviral MnSOD construct (10(9) plaque-forming units) were delivered to the tumor. Increases in MnSOD immunoreactivity and activity were seen after transduction with the adenovirus-MnSOD construct. Increasing MnSOD levels correlated with increased doubling time. Cell growth, plating efficiency, and growth in soft agar decreased with increasing amounts of the adenovirus MnSOD construct. Tumors grew slower and survival was increased in nude mice injected with the adenoviral MnSOD construct compared with the parental cell line, whereas multiple injections of the adenoviral MnSOD construct further inhibited tumor cell growth and extended survival. These results suggest that MnSOD may be a tumor suppressor gene in human pancreatic cancer. Delivery of the MnSOD gene may prove beneficial for suppression of pancreatic cancer growth.

Topics: Adenoviridae; Agar; Animals; Antioxidants; Blotting, Western; Cell Division; Cell Line, Tumor; Densitometry; Dose-Response Relationship, Drug; Gene Transfer Techniques; Humans; Mice; Mice, Nude; Mutation; Neoplasm Transplantation; Pancreatic Neoplasms; Phenotype; Superoxide Dismutase; Time Factors

The effects of 1 mM sodium butyrate or 2% dimethylsulfoxide (DMSO) on three human pancreatic tumor cell lines were examined. The cell lines tested were MIA PaCa-2, PANC-1 and CAPAN-1. Both butyrate and DMSO inhibited the ability of all three lines to form colonies in soft agar. These results suggest that the use of these agents provides a model system for the study of the molecular changes involved in human pancreatic cancer. In butyrate all the cell lines showed a marked increase in cellular levels of alkaline phosphatase, while growth in DMSO led to a reduction in most cases. DMSO caused a rapid reduction in the attachment of all three cell lines to collagen substrates, while butyrate had no effect. These results illustrate the fact that although both butyrate and DMSO appear to greatly reduce the parameters correlated with tumorigenicity of human pancreatic cancer cells, the mechanisms involved may be very different.

Topics: Agar; Alkaline Phosphatase; Amylases; Butyrates; Butyric Acid; Cell Adhesion; Cell Division; Cell Line; Clone Cells; Collagen; Dimethyl Sulfoxide; Humans; Pancreatic Neoplasms

Topics: Adenocarcinoma; Adenocarcinoma, Scirrhous; Adolescent; Adult; Agar; Aged; Carcinoma; Chromatography; Colonic Neoplasms; Gallbladder Neoplasms; Hemangiosarcoma; Humans; Liver Neoplasms; Lymphoma, Non-Hodgkin; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Pancreatic Neoplasms; Protein Denaturation; Retroperitoneal Neoplasms; Stomach Neoplasms

Topics: Acyltransferases; Agar; Bile Duct Neoplasms; Biliary Tract Diseases; Cholelithiasis; Electrophoresis; Glutamates; Hepatitis; Humans; Isoenzymes; Jaundice; Liver Cirrhosis; Liver Diseases; Liver Neoplasms; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis

Topics: Agar; Aminopeptidases; Biliary Tract Diseases; Clinical Enzyme Tests; Electrophoresis; Gels; Humans; Isoenzymes; Pancreatic Diseases; Pancreatic Neoplasms