The Expression Of HSulf-1in Malignant Melanoma And Its Role In Surpressing Tumor Cell Migration

Part I The expression of HSulf-1in the tissue microarray (TMA) of human malignant melanoma and benign nevus patientsObjective The purpose of the this study was to evaluate the protein expression of HSulf-1in the tissue biopsies of human malignant melanoma and benign nevusMethod Immunofluorescent was used to analyse HSulf-1expression in human benign nevus (N=16), primary melanoma (N=30) and metastatic melanoma (N=24) tissue samples from tissue microarray (TMA). Semi-quantitation of HSulf-1staining from TMA was performed by Image Pro Plus image software.Results The immunofluorescence results of TMAs revealed a statistically significant lower expression of HSulf-1in metastatic melanoma when compared to primary melanoma (p＜0.05). No difference was observed between benign nevus and primary or benign nevus and metastatic melanoma (p>0.05).Conclusion HSulf-1may inhibit in the processes from primary melanoma to metastatic melanoma. Part II The expression of HSulf-1in in the human normal melanocytes and melanoma cell lines.Objective The purpose of the this study was to evaluate the expression of HSulf-1in human normal melanocytes and melanoma cell lines.Method We examed the expression of HSulf-1by real-time RT-PCR, Western blotting in Human Epidermal Melanocytes-adult (HEMa) and melanoma cell lines A375. Malme-3M, SK-MEL-2、SK-MEL-5、SK-MEL-28、M14、MV3in mRNA and protein levels.Results The expression of HSulf-1was significantly higher in human normal melanocytes and primary melanoma cell lines than metastatic cell lines.Conclusion HSulf-1may inhibit in the metastatic processes of melanoma. Part Ⅲ Establishment of melanoma cells with HSulf-1over-expressionObjective The purpose of the this study was to establish a cell line expressing the HSulf-1protein in order to explore the role of HSulf-1in the metastatic processes of melanoma.Method Stable HSulf-1gene expression vector pcDNA3.1/Myc-His (-) plasmids were transfected into low melanoma cell line MV3with HSulf-1expression. Western blotting, real-time RT-PCR were used to detect the protein and mRNA expression in the HSulf-1plasmid transfected cell line.Results mRNA and protein expression of HSulf-1was higher in HSulf-1plasmid transfected MV3cell line than the control cell lines.Conclusion Transfected melanoma cell line MV3over-expressed HSulf-1. Part IV The influence of HSuIf-1on cell biological functionsObjective The purpose of the this study was to explore the role of HSulf-1on HGF mediated signaling pathway, tumor cell proliferation and migration in melanoma.Method To identify the impact of HSulf-1over-expression on HGF mediated signaling pathway, proliferation and migration of melanoma cell lines, Western blot, MTT assay and transwell migration assay were used respectively.Results Overexpression of HSulf-1in melanoma metastatic cells MV3downregulated p-ERK phosphorylation but not in a HGF modulated way, decrease their migratory ability, while did not affect cell proliferation compared with vector transfected controls.Conclusion We could speculate that HSulf-1might inactivate the ERK phosphorylation but not in a HGF modulated way, reduce migration ability of the melanoma cells, thus inhibiting tumor metastasis.