中国药理学通报2011，Vol.27Issue(6)：791-797,7.DOI:10.3969/j.issn.1001-1978.2011.06.013
蛇毒半胱氨酸蛋白酶抑制剂基因转染对小鼠黑色素瘤B16细胞蛋白质表达谱的影响
Effect of snake venom cystatin gene on the protein expression profiles of mouse melanoma B16F1 cells
摘要
Abstract
Aim To investigate the effects of stable transfected sv-cystatin gene on the protein expression profiles of mouse melanoma B16F1 cells. Methods By using two-dimensional gel electrophoresis followed by MALDI-TOF-MS and database searching, the differential protein expression profile between the B16F1/svcystatin cells which stably expressed the sv-cystatin gene and the B16F1/pcDNA3. 1 cells which was transfected with the control vector pcDNA3. 1-His C was analysed. The mRNA and protein expressions of some differentially expressed proteins were confirmed by real-time PCR and Western blot. The differentially expression genes were analyzed by a web-based DAVID bioinformatics platform. Results Out of ( 412 ± 20 ) matched spots on two-dimensional gels, 11 spots with altered expressions were found, five were up-regulated and six were down-regulated. Ten proteins were identified by MALDI-TOF-MS. These altered proteins were involved in cell metabolism, small G-protein regulation . translation , etc . The results of Western blot showed that the expressions of tumor suppressor gene NM23 and the Ran regulator RhoCDI-beta were upregulated by ( 2. 23 ±0. 34 ) and ( 1. 57 ±0. 11 ) folds,respectively. The mRNA expressions of NM23 , RhoGDI-beta and RANBPI were up-regulated whereas the expressions of eIF5A, eEF1-beta2 and MDH1 were down-regulated. These results were in accordance with the data from proteomic analysis. Analysis based on the gene ontology annotation suggested the target genes of sv-cystatin mainly located in extracellular region,plasma membrane and nucleus, and the functions of these genes involved regulation of transcription, cysteine proteases and apoptosis. Conclusions The regulation of NM23 and RhoCDI might contribute to the antitumor activities of sv-cystatin. The targets of sv-cystatin might be transcriptional factors, secreted enzymes and receptors.关键词
蛇毒/半胱氨酸蛋白酶抑制剂/黑色素瘤/小鼠/蛋白质组学/基因本体学分类
医药卫生引用本文复制引用
齐元麟,纪明开,谢群,黄清玲,林建银..蛇毒半胱氨酸蛋白酶抑制剂基因转染对小鼠黑色素瘤B16细胞蛋白质表达谱的影响[J].中国药理学通报,2011,27(6):791-797,7.基金项目
国家自然科学基金资助项目(No 30371747) （No 30371747）
福建省教育厅资助省属高校项目(No 2007F5051) （No 2007F5051）
福建医科大学青年科学基金资助项目(No FJGXQ04011) （No FJGXQ04011）
