Sox10在肿瘤增殖和转移中的作用及机制-免疫学专业论文.docxVIP

PAGE PAGE IV The Function and Mechanism of Sox10 in tumorigenesis and Metastasis Abstract The Function and Mechanism of Sox10 in tumorigenesis and Metastasis Abstract Objective: SRY-box containing gene 10 (Sox10) is a member of the high mobility group (HMG) transcription factor superfamily. We have previously proved that the expression of Sox10 was diminished in cancer cells due to promoter methylation. The aim of this study was to explore the effects and molecular mechanism of Sox10 on the development of cancer. Methods: The gene encoding Sox10 was amplified and inserted into pLVX-AcGFP-N1 vector. We employed lentivirus to elevate the expression of Sox10 in hepatocellular carcinoma HCC-LM3, esophageal carcinoma KYSE150 and colorectal carcinoma HCT116 cell lines, all of which have fully methylated and completely silenced Sox10. Expression of Sox10 was determined by quantitative reverse-transcription polymerase chain reaction and western blot. The localization of Sox10 in cancer cells was investigated by confocal laser scanning microscopy.Cell growth was measured by 3-(4,5-dimethyl-thiazol-2yl) -5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, colony formation and anchorage-independent growth assay. Cell migration was quantified in vitro using transwell chambers and wound-healing assay. In situ apoptosis assay was performed with Fluorescein FragEL? DNA Fragmentation Detection Kit. Apoptotic cells carrying DNA labeled with FITC-dUTP were observed under fluorescence microscope. For the tumorigenicity assay, HCC-LM3, KYSE150 and HCT116 cell lines infected with LV-Sox10 or LV-GFP were injected subcutaneously into the right flank of each mouse (6 mice/group) and the tumor volume was calculated. The interaction between β-catenin and Sox10 was confirmed by immunoprecipitation (IP) analysis of 293T cells transfected with β-catenin and V5-Sox10. For the luciferase reporter assay, cells were co-transfected with TOPFlash or FOPFlash luciferase report