neuropeptide-y and Carcinoma

Overexpression of neuropeptide Y (NPY) and its receptor system has been reported in various types of cancers. NPY Y5 receptor (Y5R) has been implicated in cell growth and angiogenesis. However, the role of Y5R in breast cancer is unknown. To identify the role of Y5R in breast cancer, we screened several breast cancer cell lines to examine the expression of Y5R and its function in breast cancer. All screened cell lines express both Y1 receptor and Y5R except BT-549, which expresses mainly Y5R. Binding studies showed that NPY, Y5R-selective agonist peptide, and Y5R-selective antagonist (CGP71683A) displaced (125)I-PYY binding in BT-549 cell membranes in a dose-dependent manner. The displacement studies revealed the presence of two binding sites in Y5R with IC(50) values of 29 pmol/L and 531 nmol/L. NPY inhibited forskolin-stimulated cyclic AMP accumulation with an IC(50) value of 52 pmol/L. NPY treatment of BT-549 cells induced extracellular signal-regulated kinase phosphorylation but did not alter intracellular calcium. Y5R activation stimulates BT-549 cell growth, which is inhibited by CGP71683A, pertussis toxin, and extracellular signal-regulated kinase blockade. CGP71683A alone induced cell death in a time- and dose-dependent manner in Y5R-expressing cells. The stimulation of MDA MB-231 cell migration by NPY is inhibited by CGP71683A. Together, our results suggest that Y5R plays an important role in cancer cell growth and migration and could be a novel therapeutic target for breast cancer.

Topics: Binding Sites; Binding, Competitive; Breast Neoplasms; Carcinoma; Cell Line, Tumor; Cell Movement; Cell Proliferation; Colforsin; Cyclic AMP; Dose-Response Relationship, Drug; Extracellular Signal-Regulated MAP Kinases; Female; Humans; Naphthalenes; Neoplasm Invasiveness; Neuropeptide Y; Phosphorylation; Pyrimidines; Receptors, Neuropeptide Y

The innervation of the human adrenal gland and of cortical lesions was studied in sections of cortical tissue (n=10), hyperplastic cortical tissue (n=3), and tissue from cortical adenomas (n=5) and carcinomas (n=6). The presence and distribution of nerve structures containing neuronal markers indicating sympathetic and parasympathetic innervation were studied by immunohistochemistry and the co-existence and co-localization patterns of the different markers by immunofluorescence. The cortex and hyperplastic cortical tissue had a moderate to rich supply of nerve structures containing the typical neuronal markers: protein gene product 9.5 (PGP 9.5), neuron-specific enolase (NSE), small vesicle synaptic protein type 2 (SV2), and nerves showing immunoreactivity to the adrenergic marker tyrosine hydroxylase (TH). All these immunoreactive nerves were located predominantly adjacent to blood vessels, but also among parenchymal cells. The cortex showed numerous nerve structures containing the neuropeptide substance P (SP), neuropeptide Y (NPY) and vasoactive intestinal protein (VIP), but few nerves containing these peptides were seen in hyperplastic cortical tissue. Typical markers were occasionally observed in cortical adenomas but were not found in carcinomas, except in a few cases where PGP 9.5 and NSE were present, but only adjacent to necrotic areas. Nerves containing NPY and VIP occurred in varying numbers in both adenomas and carcinomas. NPY- and VIP-immunoreactive nerve structures were seen mostly alongside blood vessels. There were several types of co-existence. For instance, NSE/VIP-, TH/VIP- and TH/NPY-immunoreactive nerve structures were often seen in the same trunk, but were only partly co-localized.

Topics: Adenoma; Adrenal Cortex; Adrenal Cortex Neoplasms; Carcinoma; Fluorescent Antibody Technique, Indirect; Humans; Hyperplasia; Immunoenzyme Techniques; Membrane Glycoproteins; Nerve Tissue Proteins; Neuropeptide Y; Parasympathetic Nervous System; Phosphopyruvate Hydratase; Sympathetic Nervous System; Thiolester Hydrolases; Tyrosine 3-Monooxygenase; Ubiquitin Thiolesterase; Vasoactive Intestinal Peptide

Choroid plexus carcinoma is a rare neoplasm derived from the epithelium of the choroid plexus. The production and secretion of endothelin-1 (ET-1) by cultured human choroid plexus carcinoma cells were studied by radioimmunoassay and Northern blot analysis. Immunoreactive (IR)-ET was detected in the culture medium (2.78 +/- 0.12 fmol/10(5) cells/24 h; n = 5; mean +/- SEM) but not in the unconditioned medium. Reverse-phase high-performance liquid chromatography of the extract of the culture medium showed a single peak eluting in the position of ET-1. Treatment with tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) or a combination of interferon-gamma (IFN-gamma), TNF-alpha, and IL-1 beta caused significant increases in the IR-ET levels in the culture medium. Northern blot analysis of total RNA showed the expression of ET-1 mRNA in choroid plexus carcinoma cells. The expression levels of ET-1 mRNA were increased by treatment with a combination of IFN-gamma, TNF-alpha, and IL-1 beta. The present study has shown the production and secretion of ET-1 by cultured human choroid plexus carcinoma cells and suggests the possibility that ET-1 formation is related to the pathophysiology of this tumor.

Topics: Blotting, Northern; Calcitonin Gene-Related Peptide; Carcinoma; Choroid Plexus Neoplasms; Chromatography, High Pressure Liquid; Cytokines; Endothelin-1; Humans; Neuropeptide Y; Radioimmunoassay; Tumor Cells, Cultured

Neuropeptide Y (NPY) is a 36 amino acid peptide known to inhibit glucose-stimulated insulin secretion. NPY has recently been shown to be synthetized within rat islets of Langerhans and to be secreted in a differentiated rat insulin-secreting cell line, and as to this date the localization of NPY in human endocrine pancreas has not been reported. As NPY shares high amino acid sequence homology with peptide YY (PYY) and pancreatic polypeptide (PP), the polyclonal antibodies raised against these peptides often cross-react with each other. To demonstrate the presence of NPY in the human endocrine pancreas, we used a highly specific monoclonal antibody raised against NPY and another against its C-flanking peptide (CPON). We studied three cases of hyperplasia of Langerhans islets and 11 cases of endocrine tumors of the pancreas. NPY and CPON were detected in all three cases of hyperplasia. For the 11 pancreatic tumors, five and nine of the tumors were positive for the antibodies NPY and CPON, respectively. The two negative tumors for CPON immunoreactivity were differentiated insulinomas, which showed no evidence of other hormonal secretion. In normal Langerhans islet, NPY and CPON immunoreactivities were colocalized in glucagon-producing cells (alpha-cells) and in a few insulin-secreting cell (beta-cells).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Animals; Antibodies; Carcinoid Tumor; Carcinoma; Cell Line; Female; Humans; Hyperplasia; Immunohistochemistry; Insulin; Insulinoma; Islets of Langerhans; Male; Middle Aged; Neuroendocrine Tumors; Neuropeptide Y; Pancreatic Hormones; Pancreatic Neoplasms; Rats

Twenty-two neuroendocrine tumours of the larynx were investigated using a panel of immunocytochemical markers. Three were small cell carcinomas, eight were large cell neuroendocrine carcinomas and 11 were paragangliomas. Twenty were positive for protein gene product 9.5, 19 for neuron-specific enolase, 15 for chromogranin A, nine for bombesin, eight for substance P, eight for neuropeptide Y, eight for metenkephalin, seven for somatostatin, five for calcitonin, eight for calcitonin gene-related peptide and one for vasoactive intestinal polypeptide. Bombesin immunoreactivity was largely restricted to the small cell carcinomas and large cell neuroendocrine carcinomas and neuropeptide Y, metenkephalin and substance P to the paragangliomas. This comprehensive immunocytochemical analysis of neuroendocrine tumours of the larynx demonstrates that these tumours represent special entities but have similar patterns of immunostaining to those of neuroendocrine tumours in other sites.

Topics: Adult; Aged; Aged, 80 and over; Bombesin; Calcitonin; Carcinoma; Carcinoma, Small Cell; Chromogranin A; Chromogranins; Enkephalin, Methionine; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Neuropeptide Y; Neuropeptides; Neurosecretory Systems; Paraganglioma; Phosphopyruvate Hydratase; Somatostatin; Substance P

Cervical carcinoma cell lines produce several polypeptide hormones. While screening human cervical cancers for such factors we have found many tumors to be immunoreactive for opioid peptides. The tumors were collected at surgery or prior to actinotherapy, fixed in buffered formalin, paraffin-embedded, and immunocytochemically stained using the PAP method. The localization of opioid-like immunoreactivity was investigated using a primary antiserum recognizing the Tyr-Gly-Gly-Phe sequence common to all known opioid peptides. The presence of neuropeptide Y (NPY) and peptide histidine methionine (PHM), both known to be present in normal human cervical tissue, was also investigated using specific antisera. Controls included staining controls and absorption controls, using Sepharose-coupled antigen and were all negative. Out of 40 cervical carcinomas, 27 displayed varying numbers of opioid-immunoreactive cells. In 12 normal cervical specimens no specific staining was observed. None of the tumors displayed any NPY-like or PHM-like immunoreactivity. The growth of cervical carcinoma seems therefore to be accompanied by a destruction of the local peptidergic innervation of blood vessels and smooth muscle cells. The demonstration of opioid-like immunoreactants and the known immunoregulatory properties of these peptides, indicate that opioid peptides may be important in the regulation of growth of human cervical carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma; Endorphins; Female; Humans; Immunohistochemistry; Middle Aged; Neuropeptide Y; Peptide PHI; Uterine Cervical Neoplasms

In order to develop an immunoradiometric assay for human neuropeptide (hNPY), a recently discovered and potent vasoconstrictor 36-amino-acid peptide, we used hNPY and some of its subpeptides to prepare monoclonal anti-NPY antibodies. Two monoclonal antibodies with high affinity for hNPY, that is affinity constants in the range of 10(10) mol/L-1, which respectively, reacted with the 9-18 portion and the 32-36 portion of hNPY were used in the immunoradiometric system. The assay was highly specific, NPY-related peptides such as pancreatic polypeptide and peptide YY not being detected. The lower limit of sensitivity was 0.5 pmol/L. In 303 normal subjects, plasma NPY concentrations were less than 0.5 pmol/L in 67%, 0.5 to 5.0 pmol/L in 25% and 5.1 to 30 pmol/L in the remaining 8%. A value of 7.5 pmol/L (95th percentile value in the normal group) was considered as the upper limit of normal. Among 111 patients with various neuroendocrine tumors, elevated plasma NPY concentrations were found in patients with pheochromocytomas and neuroblastomas, the highest plasma levels being found in patients with malignant pheochromocytomas. We conclude that patients with neuroendocrine tumors, especially secreting and or malignant tumors of the sympathochromaffin system, often have elevated plasma NPY concentrations.

Topics: Adolescent; Adult; Aged; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoid Tumor; Carcinoma; Child; Child, Preschool; Epitopes; Female; Humans; Infant; Infant, Newborn; Middle Aged; Neuroblastoma; Neuropeptide Y; Pheochromocytoma; Radioimmunoassay