lprG基因表达上调对THP1细胞表达谱的影响

Effect of high lprG gene expression on global gene expression profile in THP1 cells

目的分析lprG基因表达上调对THP1细胞表达谱的影响。方法建立稳定表达lprG的THP1细胞系,通过q PCR及Western印迹法检测lprG的表达。用RNA-seq分析稳定表达lprG对THP1细胞基因表达谱有何影响。对测序数据进行分析,作GO及KEGG通路的显著性富集分析,并选取6个差异表达基因进行q PCR验证。结果成功构建稳定表达lprG的THP1细胞系。RNA-seq检测发现共有712个差异表达的基因,其中419个上调,占58.8%,293个下调,占41.2%。富集分析显示,lprG的异位表达可影响THP1细胞多种生理或代谢过程。q PCR结果与RNAseq结果一致。结论 lprG会影响THP1细胞内与Toll样受体信号通路、补体、细胞吞噬、溶酶体及过氧化物酶体等过程相关的基因的表达,这可为进一步研究lprG与巨噬细胞相互作用机制提供前期基础。

Objective To investigate the effect of high lprG gene expression on the global geneexpression profile in THP1 cells. Methods The THP1 cell lines with stably expressing lprG wereestablished, the expression of lprG was detected by qPCR and Western blotting. The expression profileof THP1 cells with stably expressing lprG was examined with RNA.seq. The pathway enrichmentanalysis of the sequencing data was performed with Gene Ontology （GO） and Kyoto Encyclopedia ofGenes and Genomes （KEGG）, and six expressed genes were verified with qPCR. Results The THP1cell lines with stably expressing lprG. were successfully established. Total 712 differentially expressedgenes were detected by RNA-seq, of which 419 were up.regulated accounting for 58.8% and 293 weredown.regulated accounting for 41.2%. The data of GO and KEGG pathway showed that the ectopicexpression of lprG may influence the physiological process or metabolism in THP1 cells. The results ofqPCR were consistent with those of RNA-seq. Conclusion The high expression of lprG gene in THP1cells involves in many basic biological processes, including toll.receptor signaling pathway,complement, cytophagy, lysomes and peroxisome, which may provide basis for further study on theinteraction of lprG and macrophages.