KISS1 Protein Expression Research Articles

Effects Of Exercise On The Expression Of Kiss-1/GPR54 In Testis Of High-fat Diet Rats In Growth Phase

PURPOSE: To explore the effect of exercise on localization and expression of KISS-1 and GPR54 in the growth period (PND 21st to 56th day) of high-fat diet rat testes. METHODS: 21D old weanling rats were randomly divided into group HC (n=32) and group HE (n=32). HE group took 5-weeks trained (60-70% vVO2max, 1h/day, 5days/week). Groups HC and HE were fed with high-fat feed, which was purchased from Beijing Huafukang Biotechnology Co. Ltd. (LOT No: D12451).The rats of two groups were killed on the 21st D, 35th D, 43rd D, and 56th D old. The localization, mRNA expression and protein expression of KISS-1/GPR54 in the testis of each group were tested. RESULTS: The high-fat diet intervention resulted in a decrease in testicular index from pre-puberty(p<0.05); testicular tissue testosterone decreased significantly from prepuberty(p<0.01), and endocrine function was impaired; rat testicular tissue structure was loose, and the number of mature sperm decreased. The high-fat diet intervention had no significant effect on the localization of KISS-1/GPR54 system in rat testis, but affected the expression of KISS-1/GPR54 in rat testis and down-regulated the expression of KISS-1 protein(p<0.01). Exercise intervention can improve testicular development in rats with high-fat diet, especially to improve sex hormone disorders caused by high-fat diet. Under exercise intervention, the testicular tissue testosterone decline caused by high-fat diet was improved(p<0.05). Comparing with HC group, the expression of GPR54 protein increased significantly(p<0.01),but the KISS-1 protein expression had no obvious change(p>0.05). CONCLUSION: High-fat diet can inhibit the testicular development of male rats in the growth phase, and can also down-regulate the protein and gene expression levels of KISS-1/GPR54 system in testis tissue, and change the expression of KISS-1/GPR54 system, and its role needs further study. 60~70% VO2max moderate-intensity aerobic exercise change the inhibitory effect of high-fat diet on testicular development in male rats, and up-regulate KISS-1/GPR54 in prepubertal stage and whether KISS-1/GPR54 in testicular tissue participates in its regulation remains to be further study.

MicroRNA-124 and microRNA-378 inhibit the proliferation and invasion of colorectal cancer by upregulating KiSS1.

BackgroundThe KiSS1 gene is considered a tumor suppressor in various cancers. MicroRNAs are involved in many important life processes, and their regulation of gene expression may be as important as that of transcription factors. Here, we explore the roles of miR-124-3p and miR-378-3p in colorectal cancer and their relationships with the KiSS1 gene.MethodsThe effects of miR-124-3p and miR-378-3p on KiSS1 protein expression were observed by transfecting colorectal cancer cells (SW-480) with miR-124-3p and miR-378-3p mimics and inhibitors. Moreover, cell proliferation, migration and invasion were evaluated by ethynyl-20-deoxyuridine and Transwell experiments.ResultsThe KiSS1 mRNA and protein expression levels were significantly increased in mimic-transfected cells compared with those in untransfected cells, and the proliferation, migration and invasion abilities of the former were decreased; in addition, opposing results were obtained in the inhibitor and mimic groups.ConclusionsIn conclusion, our studies indicate that miR-124-3p and miR-378-3p upregulate the expression of KiSS1 and are associated with colorectal cancer metastasis and progression. miR-124-3p, miR-378-3p and KiSS1 may play important roles in colorectal cancer.

MiR-506 Targets UHRF1 to Inhibit Colorectal Cancer Proliferation and Invasion via the KISS1/PI3K/NF-κB Signaling Axis.

BackgroundThe UHRF1 gene is an epigenetic modification factor that mediates tumor suppressor gene silencing in a variety of cancers. Related studies have reported that UHRF1 can inhibit the expression of the KISS1 gene. However, the regulatory mechanism underlying UHRF1 expression in colorectal cancer (CRC) is still unclear. The aim of this study was to gain a better understanding of the regulation of UHRF1 expression in CRC and to determine whether it regulates the mechanism by which KISS1 promotes CRC metastasis.MethodsIn the present study, the levels of miR-506, UHRF1 and KISS1 expression in CRC tissues and in human CRC cell lines were studied using quantitative real-time PCR (qRT-PCR) and Western blotting. Cell proliferation, migration, and invasion assays are used to detect cell proliferation, migration, and invasion. A dual-luciferase reporter system was used to confirm the target gene of miR-506.ResultsThis study found that UHRF1 protein is highly expressed in CRC tissues and negatively correlated with KISS1 protein expression. UHRF1 overexpression activates the PI3K/NF-κB signaling pathway by inhibiting the mRNA expression levels of pathway mediators. Bioinformatics analysis and luciferase reporter gene assays confirmed that miR-506 targets UHRF1.ConclusionThis study identified the regulation of UHRF1 expression in CRC and the mechanism of CRC metastasis. UHRF1 may be a new potential target molecule for future CRC metastasis treatment.

KISS1 protein expression is associated with worse prognosis in osteosarcoma patients: a long-term follow-up study

Background: A long-term follow-up study was used to analyze the relationship between KISS1/GPR54 protein expression and the clinical prognosis of osteosarcoma (OS) patients. Methods: Forty-four paraffin-embedded OS samples conserved during the period from 2005 to 2009 were studied. Information about gender, age, pathological type, surgical stage, and clinical outcome was collected from the medical records. Results: This study included 25 males (56.81%) and 19 females (43.19%) with a median age at diagnosis of 18 years (range, 12–74 years). The follow-up duration ranged from 97 to 162 (121.36±15.46) months. The overall survival and metastatic rates were 43.18% and 65.91% respectively. For KISS1, the rate of distal metastasis and mortality in positive expression patients were 85.00% (17/20) and 75.00% (15/20) respectively, and 50.00% (12/24) and 41.67% (10/24) in negative expression patients respectively. In terms of survival time, KISS1 positive and negative patients were 55.35±14.29 and 97.46±13.06 months, respectively. P 0.05 in these three aspects. Conclusions: Expression of KISS1 protein correlated with higher distal metastasis and shorter survival time in OS patients after an average of 10.11 years of follow-up. GPR54 protein did not affect the prognosis when expressed alone.

KISS1 protein expression is associated with worse prognosis in osteosarcoma patients: a long-term follow-up study.

BackgroundA long-term follow-up study was used to analyze the relationship between KISS1/GPR54 protein expression and the clinical prognosis of osteosarcoma (OS) patients.MethodsForty-four paraffin-embedded OS samples conserved during the period from 2005 to 2009 were studied. Information about gender, age, pathological type, surgical stage, and clinical outcome was collected from the medical records.ResultsThis study included 25 males (56.81%) and 19 females (43.19%) with a median age at diagnosis of 18 years (range, 12–74 years). The follow-up duration ranged from 97 to 162 (121.36±15.46) months. The overall survival and metastatic rates were 43.18% and 65.91% respectively. For KISS1, the rate of distal metastasis and mortality in positive expression patients were 85.00% (17/20) and 75.00% (15/20) respectively, and 50.00% (12/24) and 41.67% (10/24) in negative expression patients respectively. In terms of survival time, KISS1 positive and negative patients were 55.35±14.29 and 97.46±13.06 months, respectively. P<0.05 was considered statistically significant in all three of the above aspects. For GPR54, the rate of metastasis and mortality of positive patients were 66.67% (24/36) and 61.11% (22/36), respectively, and 62.50% (5/8) and 37.50% (3/8) in negative expression patients. The survival time of GPR54 positive and negative expression patients were 73.97±11.81 and 92.75±15.42 months, respectively; however, P>0.05 in these three aspects.ConclusionsExpression of KISS1 protein correlated with higher distal metastasis and shorter survival time in OS patients after an average of 10.11 years of follow-up. GPR54 protein did not affect the prognosis when expressed alone.

Kisspeptin and Cancer: Molecular Interaction, Biological Functions, and Future Perspectives.

Cancer disease is the second leading cause of death in the world and one of the main fields of medical research. Although there is now a greater understanding of biological mechanisms of uncontrolled cell growth, invasiveness and metastasization, the multi-step process of cancer development and evolution is still incompletely understood. The inhibition of molecules activated in cancer metastasization is an hot topic in cancer research. Among the known antimetastatic genes, KiSS-1 is involved in the metastatic cascade by preventing growth of metastasis. Moreover, loss of KiSS-1 protein expression by tumor cells has been associated with a more aggressive phenotype. KiSS-1 gene encodes a 145-amino acid protein, which following proteolytic cleavage, generates a family of kisspeptins (Kp-10, -13, and -14), that are endogenous agonists for the G-protein-coupled receptor (GPR54). The antitumor effect of KiSS-1 was primarily associated with the inhibition of proliferation, migration and cell invasion and, consequently, the reduced formation of metastasis and intratumoral microvessels. In this review, we highlight the latest data on the role of kisspeptin signaling in the suppression of metastasis in various cancer types and the use modulators of KiSS/GPR54 signaling as potential novel therapeutic agents for the treatment of cancer.

Immunohistochemical expression pattern of metastasis suppressor KISS-1 protein in adenomyosis lesions and normal endometrium

ObjectiveKisspeptins are multifunctional peptides; it has been shown that they act as inhibitors of tumor metastasis in a range of cancers and that they are also involved in cell invasion through regulation of matrix metalloproteinases (MMPs). The aim of this study was to investigate the expression of KISS-1 protein in adenomyosis lesions compared with matched eutopic endometrium, as well as with endometrium from patients without adenomyosis. Study DesignIn this comparative, non-interventional study, adenomyosis and corresponding eutopic endometrium samples from women with histologically proven adenomyosis after hysterectomy, and eutopic endometrium samples from women without adenomyosis were analysed. Expression of KISS-1 protein was analyzed immunohistochemically in formalin-fixed, paraffin-embedded adenomyotic tissue specimens (n=29), matched eutopic endometrium from the same patients (n=29) and normal endometrium from patients without adenomyosis (n=29). ResultsUsing a semi-quantitative immunohistochemical score, we found that KISS-1 protein expression was higher in the adenomyotic as compared with matched eutopic glandular endometrium (p<0.05), in which in turn KISS-1 protein expression was higher than those from patients without adenomyosis (p<0.001). The inverse correlation was found in the stroma, between adenomyosis lesions and matched eutopic endometrium (p<0.01), while no statistically significant correlation was found in KISS-1 protein expression in the stroma between patients with and without adenomyosis. ConclusionsKISS-1 protein expression appears to be up-regulated in adenomyotic as compared with eutopic glandular endometrium of patients with, as well as women without adenomyosis. These findings are suggestive of the involvement of KISS-1 protein in the pathogenesis and maintenance of adenomyosis. Future studies should investigate whether KISS1 protein could be used as a marker for early and minimally invasive detection of adenomyosis, based on its differential protein expression pattern in the eutopic endometrium of patients with and without adenomyosis.

Immunohistochemical expression pattern of metastasis suppressors KAI1 and KISS1 in endometriosis and normal endometrium

ObjectiveTo analyze the expression pattern of metastasis suppressors KAI1 and KISS1 in the endometrium of patients with and without endometriosis. Study designIn this pilot study, tissue samples were prospectively collected from 38 patients with endometriosis and 29 without endometriosis, undergoing operative laparoscopy in the proliferative phase of the menstrual cycle; diagnosis or absence of endometriosis was confirmed histologically. Protein expression of KAI1 and KISS1 were analyzed immunohistochemically in endometriotic lesions and the eutopic endometrium of patients with endometriosis and without endometriosis. ResultsKAI1 expression was significantly decreased in the glandular eutopic endometrium of endometriosis patients as compared with that of patients without endometriosis (p=0.008). On the other hand, in endometriosis patients, KAI1 expression was significantly increased in the ectopic as compared with the eutopic endometrial stroma (p=0.021). There were no other significant differences in KAI1 expression between different groups. KISS1 expression in the ectopic glandular endometrium was significantly increased as compared with the eutopic glandular endometrium from patients with (p=0.004) and without endometriosis (p=0.008). There was no significant difference in KISS1 protein expression in the stromal endometrium between the three groups. ConclusionsKAI1 and KISS1 are implicated in the pathogenesis and maintenance of endometriosis. Future studies should investigate whether KAI1 and KISS1 could be used as markers for early and minimally invasive detection of endometriosis based on their differential protein expression pattern in the eutopic endometrium of patients with and without endometriosis.

KiSS1 inhibits growth and invasion of osteosarcoma cells through inhibition of the MAPK pathway

As a metastasis suppressor, KiSS1 has been implicated in numerous human cancers. However, recent studies have demonstrated that KiSS1 promotes tumor growth and metastasis in breast cancer, and it is unclear about the expression and function of KiSS1 in human osteosarcoma (OS). The aim of the present study was to investigate the role and molecular mechanisms of KiSS1 in human OS. The expression of KiSS1 was assessed by immunohistochemical assay using a tissue microarray procedure in forty cases of OS tissues. A gain-of-function approach was used to observe the effects of lentiviral vector-mediated overexpression of KiSS1 (Lv-KiSS1) on the biological behaviors including proliferative activities and invasive potential of OS MG-63 cells, indicated by MTT and Transwell assays, respectively. The results showed that the expression of KiSS1 protein in OS tissues was significantly lowered compared to that in adjacent non-cancerous tissues (42.5% vs 70.0%, P=0.023), and had negative correlation with distant metastases of the tumor (P=0.019). Overexpression of KiSS1 inhibited proliferation and invasion of OS cells with the decreased expression of p38 MAPK and matrix metalloproteinase-9 (MMP-9). Taken together, our findings indicate that the decreased expression of KiSS1 is correlated with distant metastasis of OS, and KiSS1 may function as a tumor suppressor in OS cells through inhibition of the MAPK pathway, suggesting that KiSS1 may serve as a potential therapeutic target for the treatment of cancer.

Loss of the metastasis suppressor gene KiSS1 is associated with lymph node metastasis and poor prognosis in human colorectal cancer

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.

KiSS-1 methylation and protein expression patterns contribute to diagnostic and prognostic assessments in tissue specimens for colorectal cancer

KISS1 is a metastasis suppressor lost in several solid malignancies. We evaluated the clinical relevance of KiSS-1 methylation and its protein expression in colorectal cancer. The epigenetic silencing of KiSS-1 by hypermethylation was tested in colon cancer cells (n = 5) before and after azacytidine treatment. KiSS-1 methylation was evaluated by methylation-specific PCR in colorectal cancer cells, and normal, benign, and tumor tissues (n = 352) were grouped in a training set (n = 62) and two independent validation cohorts (n = 100 and n = 190). KiSS-1 protein expression was analyzed by immunohistochemistry on tissue arrays. KiSS-1 hypermethylation correlated with transcript and protein expression loss, being increased in vitro by azacytidine. Methylation rates were 53.1, 70.0, and 80.0% in the training and validation sets, respectively. In the training set, KiSS-1 methylation rendered a diagnostic accuracy of 72.7% (p = 0.002). Combination of KiSS-1 methylation and serum CEA (p = 0.001) increased the prognostic utility of CEA alone (p = 0.022). In the first validation set, KiSS-1 methylation correlated with tumor grade (p = 0.011), predicted recurrence (p = 0.009), metastasis (p = 0.004), disease-free (p = 0.034), and overall survival (p = 0.015). In the second validation cohort, KiSS-1 methylation predicted disease-specific survival (p = 0.030). In the training set, cytoplasmic KiSS-1 expression was significantly higher in nonneoplastic biopsies as compared to colorectal tumors (p < 0.0005). In the validation set, loss of cytoplasmic expression correlated with tumor stage (p = 0.007), grade (p = 0.035), recurrence (p = 0.017), and disease-specific survival (p = 0.022). KiSS-1 was revealed epigenetically modified in colorectal cancer. The diagnostic and prognostic utility of KiSS-1 methylation and expression patterns suggests their assessment for the clinical management of colorectal cancer patients.

Kiss-1 gene expression after radiation and its association with proliferation and apoptosis in colorectal cancer cells

To investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells. SW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate. Compared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05). Radiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.

Expression of KiSS-1 in Epithelial Ovarian Cancer and its Role in Metastasis

Objectives To study expression of KiSS-1 and its role in migration and invasion of ovarian cancer (OC). Methods Expression of KiSS-1 was detected in tissue of 46 cases of OC and 17 cases of benign ovarian neoplasm by immunohistochemistry examination. Human OC cell line HO8910 was transfected by pcDNA3-KiSS-1 vector. The cell proliferation and invasion properties were detected by RT-RCR, MTT, clone formation rate and Boyden Chamber invasion assay. Results (1) Immunostaining showed that expression of KiSS-1protein was significantly higher in OC than that in benign ovarian tumor (P&lt;0.05). (2) KiSS-1 expression was significantly higher in cases of advanced stage and with lymphatic metastasis (P&lt;0.05). KiSS-1 expression was significantly lower in clear cell cancer compared with other histologic types (P&lt;0.05). (3)KiSS-1 gene was successfully integrated into the genomic DNA of ovarian cancer cell line HO8910. Boyden Chamber invasion assay revealed that the number of cells invading through the Matrigel filter was significantly decreased in the transfected group compared with the non-transfected. No differences were observed in cell proliferation between the two groups. Conclusion There was over expression of KiSS-1 in OC compared with that in benign ovarian tumor. The KiSS-1 gene could suppress HO8910 invasion in vitro. To elucidate the contradiction effects of metastasis suppressor genes KiSS-1in vivoandin vitroneeds deeper research.

Clinical significance of KISS1 protein expression for brain invasion and metastasis

Metastases to the brain represent a feared complication and contribute to the morbidity and mortality of breast cancer. Despite improvements in therapy, prognostic factors for development of metastases are lacking. KISS1 is a metastasis suppressor that demonstrates inhibition of metastases formation in several types of cancer. The purpose of this study was to determine the importance of KISS1 expression in breast cancer progression and the development of intracerebral lesions. In this study, we performed a comparative analysis of 47 brain metastases and 165 primary breast cancer specimens by using the antihuman KISS1 antibody. To compare KISS1 expression between different groups, we used a 3-tier score and the automated score computer software (ACIS) evaluation. To reveal association between mRNA and protein expression, we used quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. Significance of immunohistochemistry stainings was correlated with clinicopathological data. We identified that KISS1 expression is significantly higher in primary breast cancer compared with brain metastases (P < .05). The mRNA analysis performed on 33 selected ductal carcinoma brain metastatic lesions and 36 primary ductal carcinomas revealed a statistically significant down-regulation of KISS1 protein in metastatic cases (P = .04). Finally, we observed a significant correlation between expression of KISS1 and metastasis-free survival (P = .04) along with progression of breast cancer and expression of KISS1 in primary breast cancer specimens (P = .044). In conclusion, our study shows that breast cancer expresses KISS1. Cytoplasmic expression of KISS1 may be used as a prognostic marker for increased risk of breast cancer progression.

Abstract 1439: Clinical significance of KISS1 protein expression in breast cancer metastasis to brain

Abstract Aim: The present study was aimed to investigate whether KISS1 suppressor protein could be used as a prognostic marker in development of breast cancer metastases. Methods: To test whether breast cancer brain metastases (BMHB) have altered expression of KISS1, we created a TMA (tissue microarray) recipient block containing 136 specimens: 103 specimens of primary breast cancer cells (invasive ductal carcinoma, IDC), 7 specimens from draining lymph nodes (LN), 39 brain metastases and 14 samples of normal breast tissue (NBreast). Each core was duplicated. Sections from this block were stained with antibodies that recognize KISS1. Immunoassays were graded based on: A) intensity of immunoreactive cells staining (Negative, 0% of the cells have staining; 1+, 5-20% of total cells have staining (Mild); 2+, 20-50% of total cells have staining (Moderate); 3+, 50% or more of total cells have staining(High)) and B) score calculated by ACIS system (Chromavision/Clarient, Aliso Viejo, CA). Positive staining in the present study was defined as visibility at low magnification (10x) and was confirmed by two pathologists. To validate the IHC findings, relative expression of KISS1 was evaluated by real time PCR technique. Results: Based on TMA expression results, we found that breast cancer metastases exhibited statistically significant less KISS1 expression vs. primary specimens (Kruskal-Wallis p value = 0.011). In validation set, 84.2% of IDC samples had either moderate (38/76) or high (26/76) expression of KISS1 protein whereas weak (negative or mild) expression was observed in 15.8% cases (12/76). In contrast, metastatic cases demonstrated weak expression in 100% cases (19/19). The real time PCR data confirmed IHC findings. We analyzed the correlation between the level of KISS1 expression and breast cancer prognosis based on the IHC data. The group of patients with high expression of KISS1 protein (1x and more) was found to have a trend toward slower disease progression. Those patients developed brain metastases 51.0+15.0 (CI: 17.7-84.3) months after tumor resection whereas the group of patients with no KISS1 expression had metastases after only 27.0+4.93 (CI: 15.39-38.69) months. Conclusions: These results suggest that high level of KISS1 protein expression is advantageous for breast cancer patients and that low levels correlate with metastases towards secondary organs such as brain. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1439. doi:10.1158/1538-7445.AM2011-1439

Correlation of KiSS-1, KiSS-1R and MMP-9 expression to invasion and metastasis of hepatocellular carcinoma and its significance

Objective To detect the expression of KiSS-1, KiSS-1R and MMP-9 in hepatocellular carcinoma (HCC). To study the correlation of KiSS-1, KiSS-1R and MMP-9 expression with invasion and metastasis of HCC, and to explore the underlying mechanisms. Methods The expression of KiSS-1 , KiSS-1R mRNA in 33 HCC samples, 26 non-neoplastic adjacent liver tissue samples and 13 non-neoplastic distant liver tissue samples were detected by RT-PCR. Tissue chips were constructed by modified manual tools, which contained HCC, non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues, normal liver tissues and intrahepatic metastasis lesions. The expression of KiSS-1 and MMP-9 protein was determined by tissue chips, immunohistochemistry and semi-quantitative image analysis in 150 HCC, 137 non-neoplastic adjacent liver tissue, 98 non-neoplastic distant liver tissues, 16 normal liver tissues and 37 intrahepatic metastasis lesion samples. Results The results of RT-PCR showed that compared with the non-neoplastic adjacent liver tissues and the non-neoplastic distant liver tissues, the expression of KiSS-1 mRNA in HCC was significantly lower (P＜0.01). The expression of KiSS-1R mRNA did not changed in HCC and non-neoplastic liver tissues (P＞0.05). The expression of KiSS-1 protein was lower in HCC with metastasis and in clinical stage Ⅲ than that in those with non-metastasis, and in clinical stages Ⅰ and Ⅱ . It was also higher in the primary than in the metastasis lesions (P＜0.01, respectively). The expression of MMP-9 was higher in tumors having peplos invasion and metastasis than in those with negative peplos invasion and non-metastasis. It was lower in the primary than the metastasis lesions (P＜0. 01, respectively).Negative correlation between KiSS-1 and MMP-9 expression was found in HCC(r=- 0.340,P＜0.01). Conclusions The imbalance between KiSS-1 and MMP-9 expression might play an important role in enhancing the invasive and metastatic capacity of HCC. Loss of KiSS-1 expression might predict an aggressive clinical behavior and was associated with metastatic potential in HCC. Key words: Liver neoplasm; KiSS-1; KiSS-1R; MMP-9; Tissue chip

Effect of anti-oncogene Kiss-1 gene on cell proliferation in human melanoma cells

Objective To investigate the effect of Kiss-1 on cell proliferation in human melanoma cancer cell,and screen a target for tumor biotherapy. Methods The eukaryotic expression vectors with Kiss-1 gene were used to transfect A375 cells by liposome-mediated transfection in vitro. The cells were selected by G418. The expression of Kiss-1 gene was confirmed by Western Blotting. FACS was used to detect the ability of cells proliferation and apoptosis. Results There were significantly high expression of Kiss-1 protein in the transfected A375 cells(1.20 ±0.21 vs control 0.60 + 0. 41), whose growth rate was significantly slower than that of control. Flow cytometry showed that apoptosis of A375 cells transfected Kiss-1 were induced(7.23% vs control 4.62%). Conclusion When transfected into A375 cells,exogenous Kiss-1 could cause the cell growth arrest to G1 ,and induce cells apoptosis. Key words: Melanoma Kiss-1; Genes,tumor suppressor,Apoptosis

A twisted kiss: in vitro and in vivo evidence of genetic variation and suppressed transcription of the metastasis-suppressor gene KiSS1 in early breast cancer

KiSS-1 is ametastasis suppressor gene, its inactivation linked to advanced tumor stage and dismal prognosis. We studied its mutational status ,transcription and protein expression in human cancer cell lines and patients with early breast cancer.<br />Tumor tissue DNA and messenger RNA (mRNA) of KiSS1 exons III and IV from the human cancer cell lines Hela, Jurkat, A549, W138t, MCF-7 and from formalin-fixed resected breast adenocarcinomas from 50 women were analysed by means of PCR-SSCP, RT-PCR and sequencing. Tumor tissue was stained for KiSS1 protein expression by means of the streptavidin-biotin complex immunoperoxidase assay. Presence of KiSS1 mutation, mRNA levels and protein staining were examined for correlations with patient/tumor characteristics. <br />A transversion in exon IVa replacing cytosine with guanine was identified 242 base pairs from the translation start site (242C>G) in the cell lines MCF-7, A549 and in 5/50 tumors (10%), resulting in substitution of proline by arginine (P81R) and alteration of the protein tertiary structure. As the substitution was present in germ-line DNA in 3/5 breast cancer patients harbouring the polymorphism in their tumor, the incidence of tumour-specific somatic mutation was 4% among the 50 patiens with early breast cancer. Although the P81R substitution was associated with reduced KiSS1 protein immunoreactivity (56% in wild-type tumors versus 20% in KiSS1-variant tumours) and with axillary nodal involvement (55% in wild-type versus 80% in KiSS1-variant tumors), the correlations did not reach statistical significance. KiSS1 mRNA was detected in only 15/48 tumours (31%) and showed no correlation with mutation or protein expression. Twenty-six tumors stained for KiSS1 protein, in contrast to the universal strong staining seen in normal breast parenchyma and placental tissues. At amedian follow-up of 38 months, relapses occurred in 20% of women with non wild-type tumors versus 13% of women with wild-type KiSS1 tumors (p=0.7). Presence of KiSS1 mutation, mRNA levels and protein expression did not have prognostic significance for relapse-free survival.<br />In conclusion, altered nucleotide sequence and repression of transcription are two potential mechanisms of suppression of the anti-metastatic effects of KiSS1 in early breast cancer: Confirmation in larger cohorts and study of functional effects of the 242C>G exon IVa mutation are warranted. KiSS1, metastasis-suppressor gene, breast cancer, mutation, transcription.

Expression of KiSS‐1 Gene and its Role in Invasion and Metastasis of Human Hepatocellular Carcinoma

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.

Effect of KISS-1 on invasive potential and proliferation of esophageal squamous carcinoma cell line EC-1

To investigate the effect of KISS-1 expression on the potential of invasion and proliferation of esophageal squamous carcinoma cell EC-1. Protein and mRNA expressions of KISS-1 were evaluated by Western blot and RT-PCR in four esophageal carcinoma cell lines (EC-1, Eca109, EC9706 and TE-1). Using liposome-mediated transfection, an eukaryotic expression vector (pcDNA3.1-KISS-1) of KISS-1 gene was transfected into EC-1 cells. Boyden chamber model, MTT and clone formation assay were used to detect the potential of invasion and proliferation. Western blot and RT-PCR showed a baseline low level of expression of KISS-1 protein (0.715 +/- 0.109) and mRNA (0.670 +/- 0.176) in EC-1 cells. pcDNA3.1-KISS-1 expression vector was successfully transfected into EC-1 cells. Western blot and RT-PCR showed that the expression of KISS-1 protein (1.143 +/- 0.218) and mRNA (0.877 +/- 0.162) in EC-1 cells transfected with pcDNA3.1-KISS-1 were significantly higher than those transfected with the control vector pcDNA3.1 (0.745 +/- 0.130, 0.685 +/- 0.128; t = 3.850, 2.481, P < 0.05) and the control cells (0.855 +/- 0.184, 0.677 +/- 0.138; t = 2.275, 2.306, P < 0.05). Boyden chamber analysis showed that the invasiveness of the cells transfected with KISS-1 at 24 h (91.8 +/- 11.7), 48 h (117.8 +/- 11.1) and 72 h (139.2 +/- 11.8) were significantly reduced than that of the cells transfected with the control vector pcDNA3.1 (118.1 +/- 14.7, 141.7 +/- 13.2, 162.2 +/- 22.7; t = 3.153, 4.215, 3.569, P < 0.01) and the control cells (112.2 +/- 15.6, 138.1 +/- 13.0, 162.3 +/- 14.0; t = 4.154, 3.797, 2.702, P < 0.05). MTT showed that the proliferation potential of cells after transfection with KISS-1 at 48 h (0.517 +/- 0.127) and 72 h (0.394 +/- 0.137) were significantly reduced than that of cells transfected with the control vector pcDNA3.1 (0.636 +/- 0.186, 0.513 +/- 0.150; t = 2.054, 2.709, P < 0.05) and the control cells (0.646 +/- 0.135, 0.511 +/- 0.153; t = 2.276, 2.205, P < 0.05). Clone formation assay suggested that cells transfected with KISS-1 (157.2 +/- 36.4) showed significantly decreased clone formation than cells transfected with the control vector pcDNA3.1 (236.3 +/- 78.1; t = 3.441, P < 0.01) and the control cells (242.5 +/- 48.6; t = 2.250, P < 0.05). KISS-1 gene inhibits the potential of invasion and proliferation of EC-1 cells.