kiss1-protein--human and Pancreatic-Neoplasms

Pancreatic adenocarcinoma (PDAC) still represents a devastating disease associated with a very limited survival. Novel biomarkers allowing an early diagnosis as well as an optimal selection of suitable treatment options for individual patients are urgently needed to improve the dismal outcome of PDAC patients. Recently, alterations of Kisspeptin serum levels, a member of the adipokine family, were described in various types of cancers. However, the role of circulating Kisspeptin as a biomarker in PDAC patients is poorly defined. In this study, we measured Kisspeptin serum levels in a cohort of 128 prospectively enrolled PDAC patients undergoing surgical resection as well as 36 healthy controls. Kisspeptin concentrations were elevated in PDAC patients compared to control samples. Nevertheless, Kisspeptin serum levels were independent of tumor-related factors such as the tumor grading, TNM stage, or clinical features such as the ECOG performance status. Finally, in our analysis, neither preoperative nor postoperative Kisspeptin levels turned out as a significant predictor of overall survival after tumor resection. In conclusion, our data suggest that Kisspeptin concentrations are altered in PDAC patients but do not allow to predict patients' outcome after resection of PDAC.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Case-Control Studies; Cohort Studies; Female; Humans; Kisspeptins; Male; Middle Aged; Pancreatic Neoplasms; Postoperative Period; Preoperative Period; Treatment Outcome

The onset of local invasion and lymphatic metastasis in pancreatic cancer limits survival following surgical intervention and additional therapies. Reduced expression of KiSS‑1 in pancreatic cancer is associated with cancer metastasis. Previous studies have indicated that kisspeptin, the KiSS‑1 peptide, is able to bind to its receptor‑GPR54 (hOT7T175) and suppress the migration of PANC‑1 pancreatic cancer cells. Whether the metastatic suppression of KiSS‑1 is dependent on the levels of GPR54 in pancreatic cancer cell lines remains unclear. Human BxPC‑3 pancreatic carcinoma cells are highly differentiated without exhibiting metastasis, however PANC‑1 pancreatic carcinoma cells are poorly differentiated and exhibit local and lymph node metastasis. Compared with primary cultured trophoblasts, BxPc‑3 and PANC‑1 cells were observed to express low levels of KiSS‑1 mRNA and protein, measured using reverse transcription‑quantitative polymerase chain reaction and western blotting, respectively. However, greater mRNA and protein expression levels of GPR54 were observed in PANC‑1 cells compared with BxPc‑3 cells. An MTT assay was used to investigate the effect of KiSS‑1 on BxPc‑3 and PANC‑1 cell proliferation. There were no significant differences in proliferation following transfection with KiSS‑1 in BxPc‑3 and PANC‑1 cells compared with the controls (P>0.05). A Transwell assay with chambers coated with Matrigel was used to evaluate the in vitro invasive ability of BxPc‑3 and PANC‑1 cells, with the invasion index of BxPc‑3 and PANC‑1 cells significantly reduced following 48 h of KiSS‑1 overexpression (P<0.05). The mRNA and protein expression levels of KiSS‑1 were significantly increased in BxPc‑3 and PANC‑1 cells 48 h subsequent to transfection with KiSS‑1 (P<0.05), while GPR54 expression was not altered (P>0.05). KiSS‑1 is a metastasis suppressor gene of pancreatic cancer, and this suppression is not dependent on the expression levels of GPR54. Therefore, KiSS‑1 is potentially a novel target for gene therapy.

Topics: Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Kisspeptins; Neoplasm Invasiveness; Pancreatic Neoplasms; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; Transfection

Identifying molecular targets for treatment of pancreatic cancer metastasis is critical due to the high frequency of dissemination prior to diagnosis of this lethal disease. Because the KISS1 metastasis suppressor is expressed at reduced levels in advanced pancreatic cancer, we hypothesized that re-expression of KISS1 would reduce metastases. Highly metastatic S2VP10 cells expressing luciferase (S2VP10L) were transfected with a FLAG-tagged version of KISS1 (KFM), KFMΔSS (with deleted secretion signal sequence), or pcDNA3 control plasmid (CP) and expression was confirmed by RTQ-PCR. SCID mice were implanted orthotopically with S2VP10L cells or transfectants and tumor growth and metastases were monitored using bioluminescence imaging. Mice with S2VP10L-KISS1 tumors developed fewer liver (98%) and lung (99%) metastases than S2VP10L. Unexpectedly, mice with S2VP10L-KFMΔSS tumors also had reduced liver and lung metastases, but had more metastases than mice with S2VP10L-KISS. KISS1 protein was found in the cytoplasm of both KFMΔSS and KISS1-expressing orthotopic tumors by immunohistochemistry. Metastases were not found in lungs of mice with S2VP10L-KISS1 tumors; whereas, KFMΔSS lung sections had regions of concentrated KISS1 staining, suggesting that secretion of KISS1 is needed to reduce metastasis significantly. These data suggest induction of KISS1 expression has potential as an adjuvant treatment for pancreatic cancer.

Topics: Adenocarcinoma; Animals; Disease Models, Animal; Gene Expression; Gene Expression Profiling; Humans; Kisspeptins; Liver Neoplasms; Lung Neoplasms; Mice; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Pancreatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Transplantation, Heterologous; Tumor Cells, Cultured; Tumor Suppressor Proteins; Xenograft Model Antitumor Assays

KiSS-1 was identified as a metastasis-suppressing gene in melanoma cells. The KiSS-1 gene product (metastin) was isolated from human placenta as the ligand of GPR54, a G-protein-coupled receptor. The role of metastin and GPR54 in tumor progression is not fully understood.. We investigated the clinical significance of metastin and GPR54 expression in pancreatic cancer. We evaluated immunohistochemical expression of metastin and GPR54 in pancreatic ductal adenocarcinoma tissues obtained from 53 consecutive patients who underwent resection between July 2003 and May 2007 at Kyoto University Hospital. In 23 consecutive patients, the plasma metastin level was measured before surgery by enzyme immunoassay.. Strong immunohistochemical expression of metastin was detected in 13 tumors (24.5%), while strong expression of GPR54 was detected in 30 tumors (56.6%). Tumors that were negative for both metastin and GPR54 expression were significantly larger than tumors that were positive for either metastin or GPR54 (p = 0.047). Recurrence was less frequent in patients who had metastin-positive tumors compared with those who had metastin-negative tumors (38.5% versus 70.0%, p = 0.04). Strong expression of metastin and GPR54 was significantly correlated with longer survival (p = 0.02). Metastin expression by pancreatic cancer was an independent prognostic factor for longer survival (hazard ratio, 2.1; 95% confidence interval, 1.1-4.7; p = 0.03), and the patients with a high plasma metastin level (n = 6) did not die after surgical resection.. Strong expression of metastin and GPR54 by pancreatic cancer is associated with longer survival. Metastin expression is an independent prognostic factor for the survival of pancreatic cancer patients. The plasma metastin level could become a noninvasive prognostic factor for the assessment of pancreatic cancer.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Female; Health; Humans; Immunohistochemistry; Kisspeptins; Male; Middle Aged; Pancreas; Pancreatic Neoplasms; Prognosis; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; Recurrence; Survival Rate; Tumor Suppressor Proteins

Metastin, which is a 54-residue peptide coded by KiSS-1 gene, is an endogenous ligand to a G-protein-coupled receptor GPR54. Metastin suppresses a malignant tumor to metastasize and regulates secretion of gonadotropine releasing hormone. Physiological action of metastin has been focused on in oncology. It is reported that less KiSS-1 gene and more hOT7T175 gene which codes GPR54 are expressed in pancreatic cancers than in normal pancreatic tissues; however, there is no study that investigates the relationship between clinicopathological characteristics and plasma metastin concentration in pancreatic cancer patients. The purpose of this study was to investigate the relationship between plasma metastin-like immunoreactive substance (LI) levels and clinical characteristics in pancreatic cancer patients. Thirty-three patients with pathologically confirmed pancreatic cancer before or just after treatments and 24 healthy volunteers were included in the study. Patients were grouped according to the International Union Against Cancer TNM classification. Plasma metastin-LI was measured by enzyme immunoassay. The plasma metastin-LI levels of cancer patients were significantly higher when compared with healthy volunteers. Significant relationship was not found between the plasma metastin-LI levels and the clinicopathological factors such as tumor size, invasion, lymph node metastasis and distant metastasis. The plasma metastin levels may be a significant biomarker to predict the presence of pancreatic cancer and could be used in pancreatic cancer screening.

Topics: Adult; Aged; Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Female; Humans; Kisspeptins; Male; Middle Aged; Pancreatic Neoplasms; Tumor Suppressor Proteins

To establish a model of pancreatic cancer in Spragu-Dawely (SD) rats, and to examine the expression level of KiSS-1mRNA in pancreatic cancer and non-cancerous pancreatic tissues in SD rats.. Dimethylbenzanthracene (DMBA) was directly implanted into the parenchyma of pancreas in SD rats (Group A), and DMBA combined with trichostatin (TSA) was implanted in the intervention group (Group B). The carcinogenesis of rats executed within 3 - 5 months in Group A and Group B were observed by HE staining and macrography. Meanwhile, the rats in the control (Group C) were executed in 5 months. The expression of KiSS-1mRNA was assayed by in situ hybridization.. (1) The incidence of pancreatic cancer in Group A within 3 - 5 months was 48.7% (18/37), including 17 cases of pancreatic ductal adenocarcinoma and 1 case of fibrosarcoma. The incidence of pancreatic cancer in Group B was 33.3% (12/36), including 11 pancreatic ductal adenocarcinoma and 1 fibrosarcoma. The maxial diameter of tumor mass in Group A was higher than that in Group B (P<0.05). (2) The positive rates of KiSS-1 mRNA in pancreatic cancer in Group A and Group B were significantly lower than those in non cancerous pancreatic tissues in Group A and Group B (P<0.01). The positive rates of KiSS-1mRNA in Group A or Group B with ductal adenocarcinoma were significantly lower than those in Group A or Group B without ductal adenocarcinoma (P<0.01). The middle or severely atypical ductal hyperplasia was observed in non-cancerous pancreatic tissues with the negative KiSS-1mRNA.. DMBA directly implanted into the parenchyma of pancreas can obtain an ideal pancreatic cancer model with high incidence in a short time. TSA may have an inhibitive effect on the carcinogenesis and the growth of pancreatic ductal adenocarcinoma in rats, and KiSS-1 may play an important role in inhibiting the invasion and metastasis of pancreatic cancer.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Pancreatic Ductal; Female; Gene Expression Regulation, Neoplastic; In Situ Hybridization; Kisspeptins; Male; Pancreas; Pancreatic Neoplasms; Proteins; Random Allocation; Rats; Rats, Sprague-Dawley; RNA, Messenger

Metastin, a post-translationally modified variant of KiSS1, was recently identified as an endogenous peptide agonist for a novel G-protein coupled receptor, hOT7T175 (AXOR12, GPR54). In this study, we analyzed the role of KiSS1 and hOT7T175 in both pancreatic cancer tissues and pancreatic cancer cell lines. Furthermore, we synthesized novel short variant forms of metastin and tested the inhibitory effect of those variants on in vitro cell functions that are relevant to metastasis. Pancreatic cancer tissues showed significantly lower expression of KiSS1 mRNA than normal tissues (p=0.018), while cancer tissues showed significantly higher expression of hOT7T175 mRNA than normal pancreatic tissues (p=0.027). In human pancreatic cancer cell lines, KiSS1 mRNA was highly expressed in 2 out of 6 pancreatic cancer cell lines, while hOT7T175 mRNA was expressed in all cell lines at various degrees. PANC-1 cells showed the highest expression of hOT7T175. Exogenous metastin did not suppress cell proliferation but significantly reduced the in vitro migration of PANC-1 cells (p<0.01). Metastin induced activation of ERK1 in PANC-1 and AsPC-1 cells. Finally, we synthesized 3 novel short variant forms of metastin, FM053a2TFA, FM059a2TFA, and FM052a4TFA. These metastin variants significantly suppressed the migration of PANC-1 cells and activated ERK1. These data suggest that the metastin receptor, hOT7T175, is one of the promising targets for suppression of metastasis, and that small metastin variants could be an anti-metastatic agent to pancreatic cancer.

Topics: Cell Line, Tumor; Cell Movement; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Enzyme Activation; Humans; Kisspeptins; Mitogen-Activated Protein Kinases; Neoplasm Invasiveness; Pancreas; Pancreatic Neoplasms; Phosphorylation; Protein Biosynthesis; Protein Isoforms; Proteins; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; Receptors, Neuropeptide; RNA, Messenger; Tumor Suppressor Proteins