kiss1-protein--human and Stomach-Neoplasms

Gastric cancer (GC) causes high morbidity and mortality in patients largely due to its invasion and metastasis. Kiss‑1 has been shown to be a metastasis suppressor in various malignancies. However, its clinical significance and biological functions in GC have not been thoroughly investigated. The present study investigated the association between Kiss‑1 expression and its methylation status and clinicopathological features in GC. Kiss‑1 expression was reduced in GC and its low expression was associated with poor histological grade, lymph node metastasis and TNM III+IV stage. Kiss‑1 overexpression in AGS GC cells significantly inhibited cell proliferation, migration and invasion in vitro. Kiss‑1 knockdown promoted the proliferation, migration and invasion of HGC‑27 cells. In summary, the data demonstrated that a low expression of Kiss‑1 played a suppressive role for the proliferation, migration and invasion of GC cells. Its expression and methylation levels were associated with the clinical progression of GC. Thus, Kiss‑1 is a potential diagnostic and prognostic marker as well as a new target for the treatment of GC.

Topics: Aged; Biomarkers, Tumor; Cell Line, Tumor; Cell Movement; Cell Proliferation; Disease Progression; DNA Methylation; Epigenesis, Genetic; Female; Gastrectomy; Gastric Mucosa; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Kisspeptins; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Promoter Regions, Genetic; Stomach Neoplasms

To investigate the expression of kisspeptin in gastric adenocarcinoma and its effect on the proliferation and migration of gastric cancer cells.. The level of kisspeptin in 50 gastric cancer tissues and their paracancerous tissues were detected by quantitative real-time PCR and Western blotting. Kisspeptin-siRNA,control-siRNA,p EGFP-N1-kisspeptin,p EGFP-N1 were separately transfected into MKN-45 gastric cancer cells. After 48-hour culture,the levels of matrix metalloproteinase-2( MMP-2),β-catenin,C-myc,MMP-9,kisspeptin were detected by Western blotting; MTT assay was used to detect the proliferation of MKN-45 cells; wound healing assay was performed to assess the migration ability of MKN-45 cells. After MKN-45 cells were treated with Wnt / β-catenin signal pathway inhibitor FH535,MTT assay and flow cytometry were used to evaluate cell proliferation ability and cell apoptosis,respectively.. Kisspeptin expression in gastric adenocarcinoma tissues was significantly lower than that of the adjacent normal tissues. The cell survival rate and migration rate of p EGFP-N1-kisspeptin group were significantly lower than those of p EGFP-N1 group. Cell survival rate and migration rate of kisspeptin-siRNA group were significantly higher than those of control siRNA group. The levels of MMP-9,MMP-2,β-catenin and C-myc in p EGFP-N1-kisspeptin group were significantly lower than those in p EGFP-N1 group. The levels of MMP-9, MMP-2, β-catenin and C-myc in kisspeptin-siRNA group were significantly higher than those in control-siRNA group. The proliferation and migration trend of gastric cancer cells treated with FH535 was similar to that of the p EGFP-N1-kisspeptin group.. The expression of kisspeptin decreases in gastric cancer tissues,and kisspeptin can interact with Wnt / β-catenin signaling pathway to inhibit the proliferation and migration of gastric cancer cells.

Topics: Adenocarcinoma; Apoptosis; beta Catenin; Cell Count; Cell Line, Tumor; Cell Movement; Cell Proliferation; Green Fluorescent Proteins; Humans; Kisspeptins; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Proto-Oncogene Proteins c-myb; RNA, Small Interfering; Signal Transduction; Stomach Neoplasms; Transfection

To identify potential markers for predicting invasion, metastasis, and prognosis of gastric adenocarcinoma (GAC).. The expressions of Slug, ZEB1 and KISS-1 were detected immunohistochemically in 261 GAC tissues and 80 normal gastric tissues.. The positivity rates of Slug, ZEB1, and KISS-1 in gastric tissues were 2.5%, 1.3%, and 87.5%, respectively, significantly different from the rates of 62.1%, 28.4%, and 41.1% in GAC tissues (P<0.05). The expression level of Slug was significantly correlated with the depth of invasion, lymph node metastasis, and pTNM stages; the positivity rates of both ZEB1 and KISS-1 were significantly correlated with the tumor grade, depth of invasion, lymph node metastasis and pTNM stages. Slug expression was positively correlated with ZEB1 expression, and KISS-1 expression was inversely correlated with Slug and ZEB1 expressions. Kaplan-Meier analysis showed that the overall survival time of patients with positive expressions of Slug and ZEB1 was significantly shorter than that of the negative patients, and the survival time of patients positive for KISS-1 was significantly longer than the negative patients. COX multivariate analysis showed that positive Slug, ZEB1 and KISS-1 protein expressions and pTNM stages were independent prognostic factors of GAC (P<0.05).. The abnormal expressions of Slug, ZEB1 and KISS-1 may contribute to the tumorigenesis of GAC and are related with lymph node metastasis, pTNM stages, and prognosis of GAC. The combined detection of Slug, ZEB1, and KISS-1 expression has an important value in predicting the progression and prognosis of GAC.

Topics: Adenocarcinoma; Disease Progression; Homeodomain Proteins; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Kisspeptins; Lymphatic Metastasis; Neoplasm Grading; Prognosis; Proportional Hazards Models; Snail Family Transcription Factors; Stomach Neoplasms; Transcription Factors; Zinc Finger E-box-Binding Homeobox 1

To investigate the function of the KISS-1 gene in gastric carcinoma cells and to explore its potential mechanism.. A KISS-1 eukaryotic expression vector was constructed and transfected into BGC-823 cells. Resistant clones were obtained through G418 selection. reverse transcription-polymerase chain reaction and western blotting were used to detect KISS-1 and matrix metalloproteinase-9 (MMP-9) expression in transfected cells. The growth of transfected cells was investigated by 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) proliferation assays, and the cells' invasive potential was analyzed by basement membrane (Matrigel) invasion assays. The anti-tumor effects of KISS-1 were tested in vivo using allografts in nude mice.. The expression level of KISS-1 mRNA and protein in BGC-823/KISS-1 transfected cells were significantly higher than in BGC-823/pcDNA3.1 transfected cells (P < 0.05) or the parental BGC-823 cell line (P < 0.05). The expression level of MMP-9 mRNA and protein in BGC-823/KISS-1 were significantly less than in BGC-823/pcDNA3.1 (P < 0.05) or BGC-823 cells (P < 0.05). MTT growth assays show the proliferation of BGC-823/KISS-1 cells at 48 h (0.642 ± 0.130) and 72 h (0.530 ± 0.164) were significantly reduced compared to BGC-823/pcDNA3.1 (0.750 ± 0.163, 0.645 ± 0.140) (P < 0.05) and BGC-823 cells (0.782 ± 0.137, 0.685 ± 0.111) (P < 0.05). Invasion assays indicate the invasive potential of BGC-823/KISS-1 cells (16.50 ± 14.88) is significantly reduced compared to BGC-823/pcDNA3.1 (20.22 ± 14.87) (P < 0.05) and BGC-823 cells after 24 h (22.12 ± 16.12) (P < 0.05). In vivo studies demonstrate the rate of pcDNA3.1-KISS-1 tumor growth is significantly slower than pcDNA3.1 and control cell tumor growth in nude mice. Furthermore, tumor volume of pcDNA3.1-KISS-1 tumors (939.38 ± 82.08 mm(3)) was significantly less than pcDNA3.1 (1250.46 ± 44.36 mm(3)) and control tumors (1284.36 ± 55.26 mm(3)) (P < 0.05). Moreover, the tumor mass of pcDNA3.1-KISS-1 tumors (0.494 ± 0.84 g) was significantly less than pcDNA3.1 (0.668 ± 0.55 g) and control tumors (0.682 ± 0.38 g) (P < 0.05).. KISS-1 may inhibit the proliferation and invasion of gastric carcinoma cells in vitro and in vivo through the downregulation of MMP-9.

Topics: Animals; Cell Line, Tumor; Cell Proliferation; Female; Humans; Kisspeptins; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Mice; Mice, Inbred BALB C; Neoplasm Invasiveness; Plasmids; Stomach Neoplasms; Transfection

Kisspeptin (Kisspeptin-54; KP-54) is a 54-amino acid peptide was originally known as metastin that was implicated in suppression of tumor metastasis and circulating kisspeptin has been proposed as a tumor marker for numerous cancers in humans. However, the plasma level of KP-54 in gastric cancer (GC) remains undetermined.. We aimed to investigate the plasma levels of KP-54 in patients with GC.. Plasma KP-54 levels were quantified with enzyme-immunoassay from blood samples of 40 patients with GC at their initial staging and 59 age-matched controls.. Plasma KP-54 levels were significantly higher in GC patients (63.3±17.9) than in controls (49.0±12.7) (p=0.000). Cut-off value for KP-54 was determined as 44 ng/ml and sensitivity, specificity, positive predictive value and negative predictive value, were 60%, 78%, 63%, and 74% respectively. Plasma KP-54 levels were not correlated with any clinicopathological features of GC patients (p>0.05).. Result of our preliminary study suggest that plasma KP-54 levels might be a useful parameter in diagnosis of GC.

Topics: Aged; Analysis of Variance; Case-Control Studies; Female; Humans; Immunoenzyme Techniques; Kisspeptins; Male; Middle Aged; Prospective Studies; ROC Curve; Stomach Neoplasms

Kiss-1 has been identified as a putative metastasis suppressor gene in various human malignancies. However, there is little information about its possible role in gastric carcinoma. In this study, we determined whether the Kiss-1 gene negatively regulates MMP-9 expression. cDNA microarray technology was used to identify the genes associated with metastasis by hepatocyte growth factor (HGF) in the gastric cancer cell lines, NUGC-3 and MKN-28. The levels of Kiss-1 RNA and protein were confirmed to be upregulated in HGF-treated gastric cancer cells. HGF induced Kiss-1 and MMP-9 production in a dose-dependent manner. In order to investigate roles of HGF signaling in tumor progression and metastasis, we measured effects of a specific MEK1 inhibitor (PD 098059) and a p38 kinase inhibitor (SB 203580) on HGF-mediated cell proliferation and MMP-9. Pretreatment with PD 098059 reduced MMP-9 and HGF-mediated cell proliferation, but increased Kiss-I expression. In contrast, SB 203580 pretreatment enhanced MMP-9 and cell prolifera-tion, but decreased Kiss-1 expression. Cotreatment of PD098059 and SB203580 increased the p38 phosphorylation stimulated by HGF. These results suggest that the HGF-mediated Kiss-1 overexpression is regulated mainly by the p38 activation and, furthermore, the activation of ERK might affect HGF-mediated Kiss-1 expression indirectly by the regulation of p38 kinase. Consistent with this result, p38 phosphorylation was strongly repressed by the knock-down of Kiss-1. Downregulation of Kiss-1 using Kiss-1 shRNA also increased in vitro cell invasion. In conclusion, Kiss-1 suppresses MMP-9 expression by activating the p38 MAP kinase signaling pathway.

Topics: Cell Line, Tumor; Cell Proliferation; Down-Regulation; Enzyme Activation; Flavonoids; Gene Expression Regulation, Enzymologic; Hepatocyte Growth Factor; Humans; Imidazoles; Kisspeptins; MAP Kinase Signaling System; Matrix Metalloproteinase 9; Neoplasm Invasiveness; Oligonucleotide Array Sequence Analysis; p38 Mitogen-Activated Protein Kinases; Pyridines; Stomach Neoplasms; Tumor Suppressor Proteins

Topics: Humans; Kisspeptins; Lymphatic Metastasis; Matrix Metalloproteinase 9; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stomach Neoplasms; Tumor Suppressor Proteins

Metastasis remains an incurable common complication in patients with gastric cancer. A variety of theories have been proposed to explain the inefficiency of the metastatic process. To compare protein expression of metastasis-related genes (nm23, KISS1, KAI1 and p53) between primary tumours and metastatic tumours may be useful in illustrating these theories.. Metastasis-related tissue microarrays (including normal tissues, primary tumours, nodal metastases and liver metastases) were constructed. The protein expression of nm23, KISS1, KAI1 and p53 in lymph node and liver metastases from advanced gastric cancer specimens was mainly examined by immunohistochemical staining in relation to primary tumours.. Immunohistochemical staining showed reduced protein expression of nm23, KISS1 and KAI1 in lymph node and liver metastases compared with primary tumours. Results for p53 were to the contrary.. Our investigations revealed a tendency of reduced protein expression of metastasis suppressor genes nm23, KISS1 and KAI1 in gastric cancer with the progress of metastasis. This means that the progression theory is an important determinant of metastatic efficiency.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Female; Gene Expression; Gene Expression Profiling; Genetic Markers; Humans; Immunohistochemistry; Kangai-1 Protein; Kisspeptins; Liver Neoplasms; Lymphatic Metastasis; Male; Middle Aged; NM23 Nucleoside Diphosphate Kinases; Nucleoside-Diphosphate Kinase; Oligonucleotide Array Sequence Analysis; Staining and Labeling; Stomach Neoplasms; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

To investigate the mRNA levels of Kiss-1 and KAI-1 metastasis suppressor genes in gastric cancer, and to explore its clinical value.. In situ hybridization was used on routinely paraffin-embedded sections of resected specimens of 49 cases with gastric cancer and 20 cases with pericancerous tissue.. The positive rates and the scores of Kiss-1 and KAI-1 mRNA in gastric cancer tissue were significantly lower than those in pericancerous tissue (P<0.01). The positive rates and scores in normal to mild-atypical hyperplasia cases were significantly higher than those in middle to severe-atypical hyperplasia of pericancerous tissue (P<0.05,P<0.01). The positive rates and scores of Kiss-1 and KAI-1 mRNA in patients with infiltrating depth T1~T2, without lymph node metastasis, with only first group of lymph node metastasis, and without distant organ metastasis were significantly higher than those in patients with infiltrating depth T3~T4, with lymph node metastasis, with second or third group of lymph node metastasis and with distant organ metastasis. A strong positive correlation was found between the expressive scores of Kiss-1 and KAI-1 mRNA in gastric cancer (r=0.53, P<0.01).. The expression of Kiss-1 and/or KAI-1 mRNA may be important biological markers of reflecting invasive and metastatic potential and prognosis in gastric cancer. The assays of Kiss-1 and/or KAI-1 mRNA expression level in benign lesions of stomach may have important clinical value for the protection and early-stage finding of gastric cancer.

Topics: Adult; Aged; Female; Humans; In Situ Hybridization; Kangai-1 Protein; Kisspeptins; Male; Middle Aged; Neoplasm Staging; RNA, Messenger; Stomach Neoplasms; Tumor Suppressor Proteins

To investigate the expression and clinical significance of KiSS- 1 and E- cadherin in gastric cardia carcinoma and the correlation between the two proteins.. The expression of KiSS- 1 and E- cadherin in 80 patients with gastric cardia carcinoma and 20 patients with normal gastric cardia epithelium was detected by immunohistochemical technique.. The expression of KiSS- 1 was negatively correlated with lymphatic metastasis and clinical stage (P < 0.05), but not correlated with the cancer differentiation (P < 0.05). The expression of E- cadherin was negatively correlated with lymphatic metastasis, clinical stage, and cancer differentiation (P < 0.05). Spearman test showed a positive correlation between KiSS- 1 and E- cadherin expression (r(s)=0.722, P < 0.05).. KiSS- 1 and E- cadherin may play important roles in inhibiting the invasion and metastasis of gastric cardia carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Cadherins; Cardia; Female; Humans; Kisspeptins; Male; Middle Aged; Neoplasm Metastasis; Stomach Neoplasms; Tumor Suppressor Proteins

KiSS-1 is a promising candidate tumor-suppressor gene and may play a key role in the metastatic cascade. The expression profile and the role of KiSS-1 in cancer progression are largely unknown in most of the cancers, including gastric cancer. In this study, KiSS-1 expression was evaluated by RNase protection assay and localization was done by in situ hybridization in 40 gastric cancers and their adjacent normal gastric mucosa. For comparison with clinicopathologic characteristics and patient prognosis, all patients were divided into 2 groups having high and low KiSS-1 expression by using the median as the cutoff value of KiSS-1 expression as determined by the RNase protection assay. Gastric cancers with low KiSS-1 had frequent venous invasion, distant metastasis and tumor recurrence. Accordingly, patients with low KiSS-1-expressing tumors had a significantly worse overall and disease-free survival. In multivariate analysis, KiSS-1 became the strongest independent prognostic factor among the conventional prognosticators for gastric cancer patients. Collectively, these findings suggest that KiSS-1 may play a crucial role in gastric cancer invasion and could be a useful target for therapeutic intervention.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Disease Progression; Disease-Free Survival; Down-Regulation; Female; Follow-Up Studies; Gene Expression Profiling; Genes, Tumor Suppressor; Humans; In Situ Hybridization; Kisspeptins; Male; Middle Aged; Neoplasm Invasiveness; Prognosis; Protein Biosynthesis; Proteins; Stomach Neoplasms; Tumor Suppressor Proteins