kiss1-protein--human and Carcinoma--Hepatocellular

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

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

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

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

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