小鼠脑微血管内皮细胞与新生隐球菌GXM作用后基因表达谱分析  

Gene expression profiling analysis of mouse brain microvascular endothelial cells incubated with glycuronoxylomannan of Cryptococcus neoformans

作　　者：聂舒[1,2] 李平[1,2] 朱红梅[1,2] 温海[1,2] 

机构地区：[1]上海长征医院皮肤病与真菌病研究所全军真菌病重点实验室 [2]第二军医大学附属长征医院皮肤科,上海200003

出　　处：《中国真菌学杂志》2015年第2期84-87,共4页Chinese Journal of Mycology

基　　金：国家自然科学基金(31470252;81271800)

摘　　要：目的探索新生隐球菌GXM能否影响脑微血管内皮细胞基因的表达,为进一步研究隐球菌嗜中枢性的分子机制提供线索。方法使用Roche Nimble Gen 12×135K小鼠基因表达谱芯片,筛选小鼠脑微血管内皮细胞系b End.3与不同浓度新生隐球菌GXM作用后差异表达的基因;结合基因本体论(Gene Ontology),使用top GO进行差异基因GO分析,结合GO语义挖掘与隐球菌侵袭中枢神经系统能力相关的差异表达基因的信息;采用荧光实时定量PCR对重要基因PIK3C2G和ADAMDEC1的表达水平变化加以验证。结果 b End.3细胞与新生隐球菌GXM作用前后基因表达对比发现,实验组GXM(90μg/m)组总共有402个基因表达上调,296个基因表达下调,GXM(180μg/m)组总共有421个基因表达上调,564个基因表达下调,细胞膜、细胞骨架、磷酸肌醇-3-激酶活化、1-磷酸肌醇-3-激酶活化、细胞紧密连接等生物过程差异表达基因较为富集;对PIK3C2G基因和ADAMDEC1基因表达水平变化进行荧光定量PCR验证,结果与芯片结果一致,基因表达水平不同程度上升,且与GXM浓度正相关。结论新生隐球菌GXM能够影响脑微血管内皮细胞基因表达,PIK3C2G基因、ADAMDEC1基因表达上调可能和隐球菌穿越血脑屏障有关。Objective To explore whether cryptococcal capsular polysaccharide GXM can influence the gene expression of cerebrovascular endothelial cells and providing clues for further study of the molecular mechanism of cryptococcus addicted to central. Methods The Mouse 12×135K Gene Expression Array manufactured by Roche NimbleGen was used to identify the differentially expressed genes of bEnd.3 cells after coculturing with GXM in different levels.TopGO method is used to analyze the differentially expressed genes.We analyze differences in gene function with the GO classification function and find information about the literature data.The changes of some important genes were found and then validated by RTPCR.Results After comparison,we got 698 differentially expressed genes,including 402 upregulated and 296 downregulated in the GXM （90 μg/ mL） group and 985 differentially genes,including 421 upregulated and 564 downregulated in the GXM （180 μg/ mL） group.Cryptococcal capsular polysaccharide GXM can regulate vascular endothelial cell gene expression.Genes relating to tight junction,cell transduction,cytoskeleton,metabolism of cell membrane componentsetc have been impacted.Considering literature and GO analysis results,we selected gene PIK3C2G and gene ADAMDEC1 and validated their expression levels by RTPCR.The results were in line with the microarray data.Conclusions Cryptococcal capsular polysaccharide GXM can influence the gene expression of cerebrovascular endothelial cells Increase in genetic PIK3C2G,ADAMDEC1 may relate to the ability of cryptococcus invading blood brain barrier.

关 键 词：脑微血管内皮细胞 新生隐球菌 GXM 基因芯片 

分 类 号：R379.5[医药卫生—病原生物学]