蛇毒半胱氨酸蛋白酶抑制剂基因转染对小鼠黑色素瘤B16F1细胞基因表达谱的影响  

Effects of Snake Venom Cystatin Gene on Gene Expression Profiles in Mouse Melanoma Cell Line B16F1

作　　者：谢群[1] 万榕[1] 林旭[1] 林建银[1] 

机构地区：[1]福建医科大学分子医学中心福建省高校感染与肿瘤重点实验室

出　　处：《癌症》2008年第7期716-722,共7页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.30371747);福建省科技重大项目资助(No.2002Y003);福建省高校创新团队培育计划资助(No.FWU-RT001)~~

摘　　要：背景与目的:本实验室已证明蛇毒半胱氨酸蛋白酶抑制剂(snake venom cystatin,sv-cystatin)具有抗肿瘤侵袭转移作用,为了进一步阐明其分子机制,本研究利用高通量的基因芯片技术探讨sv-cystatin基因转染对小鼠黑色素瘤细胞(B16F1)基因表达谱的影响。方法:分别将pcDNA3.1/sv-cystatin及空载体pcDNA3.1转染B16F1细胞,建立稳定转染的B16F1/sv-cystatin及B16F1/pcDNA3.1细胞株,提取总mRNA,应用高通量的基因芯片技术检测差异表达基因,半定量RT-PCR法分别验证5个上调和5个下调的差异表达基因。结果:B16F1细胞经转染sv-cystatin后,共检测了1218个基因,其中有45个差异表达基因,21个基因表达上调(Ratio>3),24个基因表达下调(Ratio<0.5),这些基因的功能涉及细胞粘附迁移、细胞免疫调节、增殖分化与凋亡、基因转录和细胞内信号转导等方面。10个差异表达基因的RT-PCR验证结果与基因芯片分析结果相符。结论:Sv-cystatin不仅具有抑制细胞外基质的作用,而且还具有细胞免疫调节、增殖分化与凋亡、基因转录和细胞内信号转导等多种生物学功能。BACKGROUND ＆ OBJECTIVE. Previously, we have demonstrated that snake venom cystatin （sv-cystatin） plays an important role in tumor invasion and metastasis. This study was to investigate the effects of sv-cystatin on the gene expression profile of mouse melanoma B16F1 cells. METHODS. pcDNA3.1/sv-cystatin plasmid was constructed and transfected into B16F1 using lipofectamine. Differentially expressed genes between B16F1 transfected with pcDNA3.1/sv-cystatin and pcDNA3.1 were analysed by high throughput microarray technique. Five up-regulated genes and five down- regulated genes were confirmed by semiquantitative reverse transcription- polymerase chain reaction （RT-PCR）. RESULTS. Out of 1218 transcript species, 45 showed altered expressions. 21 were up-regulated and 24 were down-regulated. These altered genes are involved in cell adhesion and migration, cell immunomodulation, proliferation, differentiation, apoptosis, as well as gene transcription and intra-cellular signal transduction. RT-PCR results of 10 alerted genes were in accordance with the microarray data. CONCLUSIONS. Sv-cystatin not only inhibits the extracellular matrix, but may also possess other diverse biological functions, including cell immunomodulation, proliferation and differentiation, apoptosis, gene transcription and intra-cellular signal transduction.

关 键 词：蛇毒半胱氨酸蛋白酶抑制剂 基因表达谱 高通量基因芯片 黑色素瘤 B16F1细胞 侵袭与转移 小鼠 

分 类 号：R739.5[医药卫生—肿瘤]