kiss1-protein--human and Liver-Neoplasms

Triple-negative breast cancer (TNBC) constitutes 10-20% of breast cancers and is challenging to treat due to a lack of effective targeted therapies. Previous studies in TNBC cell lines showed in vitro growth inhibition when JQ1 or GSK2801 were administered alone, and enhanced activity when co-administered. Given their respective mechanisms of actions, we hypothesized the combinatorial effect could be due to the target genes affected. Hence the target genes were characterized for their expression in the TNBC cell lines to prove the combinatorial effect of JQ1 and GSK2801.. RNASeq data sets of TNBC cell lines (MDA-MB-231, HCC-1806 and SUM-159) were analyzed to identify the differentially expressed genes in single and combined treatments. The topmost downregulated genes were characterized for their downregulated expression in the TNBC cell lines treated with JQ1 and GSK2801 under different dose concentrations and combinations. The optimal lethal doses were determined by cytotoxicity assays. The inhibitory activity of the drugs was further characterized by molecular modelling studies.. Global expression profiling of TNBC cell lines using RNASeq revealed different expression patterns when JQ1 and GSK2801 were co-administered. Functional enrichment analyses identified several metabolic pathways (i.e., systemic lupus erythematosus, PI3K-Akt, TNF, JAK-STAT, IL-17, MAPK, Rap1 and signaling pathways) enriched with upregulated and downregulated genes when combined JQ1 and GSK2801 treatment was administered. RNASeq identified downregulation of PTPRC, MUC19, RNA5-8S5, KCNB1, RMRP, KISS1 and TAGLN (validated by RT-qPCR) and upregulation of GPR146, SCARA5, HIST2H4A, CDRT4, AQP3, MSH5-SAPCD1, SENP3-EIF4A1, CTAGE4 and RNASEK-C17orf49 when cells received both drugs. In addition to differential gene regulation, molecular modelling predicted binding of JQ1 and GSK2801 with PTPRC, MUC19, KCNB1, TAGLN and KISS1 proteins, adding another mechanism by which JQ1 and GSK2801 could elicit changes in metabolism and proliferation.. JQ1-GSK2801 synergistically inhibits proliferation and results in selective gene regulation. Besides suggesting that combinatorial use could be useful therapeutics for the treatment of TNBC, the findings provide a glimpse into potential mechanisms of action for this combination therapy approach.

Topics: Azepines; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; Cysteine Endopeptidases; Gene Expression Regulation, Neoplastic; Humans; Indolizines; Kisspeptins; Liver Neoplasms; Phosphatidylinositol 3-Kinases; Scavenger Receptors, Class A; Sulfones; Triazoles; Triple Negative Breast Neoplasms

To study the expressions of hypoxia-inducible factor-1α (HIF-1α) and tumor metastasis suppressor gene (KISS-1) in patients with liver cancer and to analyze the correlation between HIF-1α and KISS-1 and liver cancer.. 20 normal liver tissues and 30 liver cancer tissues in our hospital were selected. The expressions of HIF-1α and KISS-1 in normal liver tissues and liver cancer tissues were detected via immunofluorescence assay. The mRNA expressions of HIF-1α and KISS-1 in normal liver tissues and liver cancer tissues were detected via reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of HIF-1α and KISS-1 in normal liver tissues and liver cancer tissues were detected via Western blotting. Differences of HIF-1α and KISS-1 expressions in normal liver tissues and liver cancer tissues were analyzed using SPSS 17.0 statistical software.. Immunofluorescence assay, RT-PCR, and Western blotting, showed that HIF-1α was highly expressed in liver cancer tissues, and its expression level was significantly higher than that in normal liver tissues. However, the expression of KISS-1 in normal liver tissues was significantly higher than that in liver cancer tissues. The results of analysis of variance showed that the differences of HIF-1α and KISS-1 expressions in normal liver tissues and liver cancer tissues were statistically significant (p<0.01).. The abnormal expressions of HIF-1α and KISS-1 are closely related to the development and progression of liver cancer, indicating that HIF-1α and KISS-1 have important research values in liver cancer, and the expressions of HIF-1α and KISS-1 can be used as the index of deterioration degree of liver cancer, providing a new clinical basis for diagnosis and treatment.

Topics: Adult; Carcinoma, Hepatocellular; Female; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Kisspeptins; Liver; Liver Neoplasms; Male; Microscopy, Fluorescence; Middle Aged

Kisspeptins, the products of the KISS1 gene have tumor suppressing and antimetastatic properties. We aimed to study KISS1 and KISS1R expression in colorectal cancer. We analyzed KISS1 and KISS1R expression using immunohistochemistry and image analysis in normal and malignant tissue samples from 111 patients with colorectal adenocarcinoma. KISS1 expression was much higher in the normal than in the malignant colonic mucosa. Regarding malignant tissues, KISS1 levels were higher in larger tumors, in stage III and IV cancers, in cancers with lymph node metastasis and in tumors located in the distal part of the large intestine. Patients with greater KISS1 levels had worse prognosis. No KISS1R expression was detected in normal or malignant tissues or in liver metastases. KISS1 expression is reduced during the malignant transformation of the colonic mucosa. However, larger and advanced colorectal cancers express more KISS1, without reaching the former normal levels, and increased KISS1 levels are associated with worse prognosis. Finally, neither the normal nor the malignant colonic epithelial cells produce KISS1R.

Topics: Aged; Biomarkers, Tumor; Cell Transformation, Neoplastic; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Intestinal Mucosa; Kisspeptins; Liver Neoplasms; Lymphatic Metastasis; Male; Prognosis; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1

To explore the impact of transcription factor 21 (TCF21) gene on proliferation, migration and apoptosis in SMMC-7721 hepatocellular carcinoma cell line and the related mechanism.. Using Lipofectamine™2000, we stably transfected pcDNA3.1⁺TCF21 and pcDNA3.1⁺ plasmids into SMMC-7721 cells of TCF21 over-expression group and empty vector group, respectively. The untreated cells were set as blank control group. The expression of TCF21 mRNA and protein were investigated by reverse transcription PCR and Western blotting. Cell proliferation ability was detected by MTT assay. Wound healing assay was used to observe cell migration ability. Annexin V-FITC/PI double labeling combined with flow cytometry was applied to determine cell apoptosis rate. Western blotting was performed to examine the expressions of KISS1, P53 and matrix metalloproteinase-9 (MMP-9).. TCF21 was over-expressed in TCF21-transfected SMMC-7721 cells. Compared with the control groups, the TCF21 over-expression group showed relatively weakened proliferation ability and significantly inhibited migration ability as well as the increased apoptosis. Western blotting demonstrated that up-regulated TCF21 raised the expressions of KISS1 and p53, and down-regulated MMP-9 level.. Tumor-suppressor gene TCF21 could inhibit the proliferation and migration of SMMC-7721 hepatocellular carcinoma cells and promote its apoptosis.

Topics: Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Movement; Cell Proliferation; Gene Expression; Gene Expression Regulation, Neoplastic; Humans; Kisspeptins; Liver Neoplasms; Matrix Metalloproteinase 9; RNA, Messenger; Transfection; Tumor Suppressor Protein p53

Cancer metastasis is a major contributor to patient death because of its systemic nature and resistance to therapeutic agents. KISS1, originally identified to be a metastasis suppressor, couples to its receptor KISS1R and plays a pivotal role in suppressing cancer metastasis. In this study, we investigated KISS1 and KISS1R expression in colorectal liver metastasis (CRLM), and analyzed their correlation with patients' clinicopathological variables, including prognosis.. Overall, 55 patients with CRLM who underwent hepatectomy between 2003 and 2013 were enrolled in this study. Immunohistochemistry was performed to evaluate the protein expression of KISS1, KISS1R, and matrix metalloproteinase-9 (MMP-9). Clinicopathological variables, including prognosis, were compared between low- and high-expressing groups of KISS1 or KISS1R. We analyzed the correlation of KISS1 or KISS1R protein expression with MMP-9.. Expression of both KISS1 and KISS1R was significantly correlated with overall survival (p = 0.0283 and p = 0.0275, respectively). The 5-year overall survival rate of the KISS1 and KISS1R low groups was 44.3 and 39.3 %, and 73.7 and 67.9 % in the high groups, respectively. Multivariate analysis revealed that KISS1 low expression was an independent prognostic factor (p = 0.037, hazard ratio 0.20). Moreover, KISS1 low-expression patients had more frequent distant metastasis (p < 0.05). Furthermore, KISS1 low-expressing tumor tissues expressed more MMP-9 protein (p = 0.034), which was mainly expressed in neutrophils at the metastatic tumor edge.. KISS1 could be a promising prognostic and therapeutic marker in CRLM. KISS1 low expression may induce high MMP-9 expression in neutrophils.

Topics: Aged; Biomarkers, Tumor; Colorectal Neoplasms; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Kisspeptins; Liver Neoplasms; Male; Matrix Metalloproteinase 9; Neoplasm Staging; Prognosis; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; Survival Rate

Cancer research is currently focused on blocking the metastatic process at its early steps. Some particularly attractive targets are metastasis suppressor genes, which control cancer cell dissemination. The aim of this study was to clarify the relationship between the expression of KiSS1, a metastasis suppressor gene, and disease progression in colorectal cancer patients. One-hundred and seventy-five patients who underwent surgery for colorectal cancer were enrolled in this study. We analyzed KiSS1 mRNA expression by real-time reverse transcription PCR in colorectal cancer tissue and paired adjacent normal mucosa. KiSS1 protein expression in early- and advanced-stage colorectal cancer samples was determined by immunohistochemical analysis. Decreased KiSS1 expression was significantly associated with lymph node metastasis and was an independent prognostic factor. Logistic regression analysis revealed that decreased KiSS1 expression was an independent risk factor for lymph node metastasis. Immunohistochemical analysis indicated that KiSS1 was highly expressed in the cell cytoplasm of early-stage colorectal cancer cells. The loss of KiSS1 appears to correlate with the progression of lymph node metastasis. An assessment of KiSS1 expression may assist in the accurate colorectal cancer diagnosis and may contribute to predict clinical outcomes.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Child; Colorectal Neoplasms; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Kisspeptins; Liver Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Peritoneal Neoplasms; Prognosis; Prospective Studies; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Survival Rate; Young Adult

To investigate the impact of expression of kisspeptin-1 (KiSS-1) metastasis-suppressor gene on the proliferative, adhesive and invasive abilities of human hepatocellular carcinoma (HCC) using an in vitro cell system.. The highly metastatic human hepatoma cell line MHCC97-H was transiently transfected with the pcDNA3.1/HisC vector expressing the KiSS-1 gene (experimental group) or the vector without the KisS-1 gene (blank control group). Untransfected cells served as the negative control group. Proliferative abilities of the three groups were assessed by flow cytometry and MTT assay. Adhesive abilities were assessed by MTT assays using matrigel and fibronectin. Invasive abilities and cell motility were assessed by chemoinvasion chamber assay using reconstituted matrigel and migration chamber assay using polycarbonate filters, respectively.. The experimental group showed significantly lower adhesion capacity to matrigel (0.257+/-0.029) than either the blank control group (0.374+/-0.016; t=-7.90345, P less than 0.01) or the negative control group (0.394+/-0.031; t=-7.22752, P less than 0.01). Similarly, the experimental group showed significantly lower adhesion capacity to fibronectin (0.292+/-0.004) than either the blank control group (0.394+/-0.010; t=-20.93138, P less than 0.01) or the negative control group (0.412+/-0.023; t=-11.31371, P less than 0.01). The experimental group also showed significantly lower numbers of cells with invasive capacity (42.40+/-1.14) than either the blank control group (66+/-1.58; t=-27.0711, P less than 0.01) or the negative control group (67.80 +/- 1.92; t=-25.4, P less than 0.01). Similarly, the experimental group showed significantly lower numbers of cells with chemotactic movement (65.80+/-1.92) than either the blank control group (93.80+/-2.28; t=-30.11750, P less than 0.01) or the negative control group (96.40+/-2.07; t=-24.19142, P less than 0.01). The experimental group showed slightly, but not significantly, lower cell proliferation (0.644+/-0.027) than either the blank control group (0.669+/-0.022; t=-1.60371, P?>?0.05) or the negative control group (0.678+/-0.027; t=-1.97828, P?>?0.05). In addition, there were no obvious differences between the three groups in the amounts of cells arrested in either the G1 phase or the S phase.. KiSS-1 overexpression suppresses the adhesion, invasion and motility, but not the proliferation, of hepatoma carcinoma cells in vitro. These findings imply that KiSS-1 might represent a promising new candidate for gene therapy against human hepatocellular carcinoma.

Topics: Apoptosis; Carcinoma, Hepatocellular; Cell Adhesion; Cell Line, Tumor; Cell Movement; Cell Proliferation; Humans; Kisspeptins; Liver Neoplasms; Neoplasm Invasiveness; 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 has been identified as a putative metastasis-suppressor gene in human melanomas and breast cancer cell lines. Although loss of KiSS-1 expression has been associated with progression and poor prognosis of various cancers, the exact role of KiSS-1 expression in HCC is not well-defined. Our study investigated KiSS-1 expression levels in HCC and its role in invasion and metastasis of human HCC. The expression levels of KiSS-1 and MMP-9 protein were determined by tissue microarray (TMA) serial sections, immunohistochemistry and semi-quantitative image analysis. All clinical and histological data obtained were subjected to statistical analysis. The expression of KiSS-1 protein in HCC and intrahepatic metastasis lesions was significantly lower (P < 0.01) when compared with non-tumor liver tissue and normal liver tissue. Multivariate analysis revealed a significant inverse correlation between KiSS-1 expression and o1 TNM stage, (F = 7.113, P < 0.01) and o2 intrahepatic metastasis (t = 2.898, P < 0.01). Loss of KiSS-1 in intrahepatic metastasis versus primary carcinomas was statistically significant (P<0.01). We also found a negative correlation between KiSS-1 and MMP-9 expression in HCC (r = -0.506, P < 0.01). We conclude that loss of KiSS-1 during HCC metastasis, along with a concomitant upregulation of MMP-9 suggests a possible mechanism for cell motility and invasion during HCC metastasis, with KiSS-1 emerging as a possible therapeutic target during HCC metastasis.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Hepatocellular; Disease Progression; Down-Regulation; Female; Gene Expression; Humans; Kisspeptins; Liver Neoplasms; Male; Matrix Metalloproteinase 9; Middle Aged; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Staging; Prognosis; Tumor Suppressor Proteins

The KiSS-1 gene has been reported to play an important role as a metastasis suppressor gene in various human malignancies. However, there is little information about its possible role in hepatocellular carcinoma (HCC). In this study, we evaluated the prognostic significance of the expression of KiSS-1 and its receptor AXOR12 in 142 HCC tissue specimens by immunohistochemistry. By using a cutoff level of 50%, immunoreactivity of KiSS-1 and AXOR12 was found in 6 (4%) and 11 (8%) HCCs. The expression of KiSS-1 and AXOR12 in HCC correlated with each other (r = 0.42, p < 0.0001) and with the expression in corresponding, surrounding liver tissue (both r = 0.35, p < 0.0001). Positive AXOR12 immunoreactivity in HCC correlated with advanced pT-stage of tumors and low tumor grading (r = 0.18, p = 0.032; r = -0.18, p = 0.029). High KiSS-1 expression in HCC had a statistically significant influence on diminished disease-free and overall survival in uni- (p = 0.006 and p = 0.002) and multivariate analysis (r = 2.874, p = 0.027 and r = 2.913, p = 0.026). In this study, we report for the first time that elevated KiSS-1 expression level in HCC correlates with worsened clinical outcome, as an independent prognostic marker for the aggressiveness of HCC.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Hepatocellular; Female; Humans; Immunohistochemistry; Kisspeptins; Liver Neoplasms; Male; Middle Aged; Multivariate Analysis; Prognosis; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; 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

Tumor metastasis-suppressor gene KiSS-1 is related to the metastasis of malignancies. However, its correlation to the metastasis, especially the formation of portal vein tumor thrombus (PVTT), of hepatocellular carcinoma (HCC) has seldom been reported. This study was to investigate the function of KiSS-1 gene in the formation of PVTT of HCC, and explore the machanism.. The expression of KiSS-1 and matrix metalloproteinase-9 (MMP-9) in 50 specimens of HCC (31 cases with PVTT and 19 cases without PVTT) and 11 specimens of PVTT was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).. The positive rates of KiSS-1 mRNA and protein were significantly lower in PVTT group and HCC with PVTT group than in HCC without PVTT group (18.2% and 16.1 % vs. 63.2%, 0% and 12.9% vs. 47.4%, P < 0.05)û the positive rates of MMP-9 mRNA and protein were significantly higher in PVTT group and HCC with PVTT group than in HCC without PVTT group (72.7% and 77.4% vs. 31.6%, 81.8% and 83.9% vs. 42.1%, P < 0.05). There was no significant difference between PVTT group and HCC with PVTT group (P > 0.05). KiSS-1 expression was negatively related to MMP-9 expression (r=-0.362 for mRNA, and r=-0.473 for protein, each P < 0.05).. KiSS-1 and MMP-9 are closely related to the formation of PVTT. KiSS-1 gene might suppress the formation of PVTT by suppressing MMP-9 synthesis.

Topics: Adult; Aged; Carcinoma, Hepatocellular; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Kisspeptins; Liver Neoplasms; Male; Matrix Metalloproteinase 9; Middle Aged; Portal Vein; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Suppressor Proteins; Venous Thrombosis

KiSS-1 has been cloned as a human metastasis suppressor gene and an orphan G-protein-coupled receptor (hOT7T175) identified as the endogenous receptor of the KiSS-1 product. In the present study, we evaluated the clinical importance of KiSS-1 and hOT7T175 gene expression in hepatocellular carcinoma (HCC).. The expression levels of KiSS-1, hOT7T175 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) messenger RNAs (mRNAs) were analyzed quantitatively by real-time reverse transcriptase polymerase chain reaction (RT-PCR) in 60 surgically resected HCCs. The KiSS-1/GAPDH and hOT7T175/GAPDH ratios of tumors were compared with clinicopathological findings.. Loss of KiSS-1 mRNA expression was not detected in HCCs. The mean KiSS-1/GAPDH ratio did not change between non-cancerous cirrhotic livers and carcinomas. On the other hand, the average hOT7T175/GAPDH ratios increased from non-cancerous livers (0.08) to carcinomas (0.48). Overexpression of KiSS-1 and hOT7T175 genes was recognized in 6 tumors, which were in an advanced stage and showed poor survival.. Overexpression of KiSS-1 and hOT7T175 genes was frequently observed and correlated with HCC progression; thus, the possibility that overexpressed KiSS-1 and hOT7T175 peptides mediate growth signals into cancer cells in HCCs is suggested.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Hepatocellular; Disease Progression; Disease-Free Survival; Female; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Heterotrimeric GTP-Binding Proteins; Humans; Kisspeptins; Liver Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Proteins; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; Receptors, Neuropeptide; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Tumor Suppressor Proteins