Int J Biol Sci 2012; 8(7):1013-1022. doi:10.7150/ijbs.3836
MicroRNA Expression Profile in RAW264.7 cells in Response to Brucella melitensis Infection
1. State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China;
2. Department of Preventive Veterinary Medicine, College of Animal Science & Technology, Shihezi University, Shihezi city, China
* Contributed equally to this work.
Zheng K, Chen DS, Wu YQ, Xu XJ, Zhang H, Chen CF, Chen HC, Liu ZF. MicroRNA Expression Profile in RAW264.7 cells in Response to Brucella melitensis Infection. Int J Biol Sci 2012; 8(7):1013-1022. doi:10.7150/ijbs.3836. Available from http://www.ijbs.com/v08p1013.htm
MicroRNA (miRNA) is small non-coding RNA with approximate 22 nt in length. Recent studies indicate that miRNAs play significant roles in pathogen-host interactions. Brucella organisms are Gram-negative facultative intracellular bacteria that cause Brucellosis. Brucella strains infect macrophages and establish chronic infection by altering host life activities including apoptosis and autophagy. Here, we report a comprehensive analysis of miRNA expression profiles in mock- and Brucella-infected RAW264.7 cells using high-throughput sequencing approach. In total, 344 unique miRNAs were co-expressed in the two libraries, in which 57 miRNAs were differentially expressed. Eight differentially expressed miRNAs with high abundance were subjected to further analysis. The GO enrichment analysis suggests that the putative target genes of these differentially expressed miRNAs are involved in apoptosis, autophagy and immune response. In particular, a total of 25 target genes are involved in regulating apoptosis and autophagy, indicating that these miRNAs may play important regulatory roles in the Brucella-host interactions. Furthermore, the interactions of miR-1981 and its target genes, Bcl-2 and Bid, were validated by luciferase assay. The results show that miR-1981 mimic up-regulated the luciferase activity of psiCHECK-2 Bcl-2 3′ UTR, but the luciferase activity of psiCHECK-2 Bid 3′ UTR was not changed significantly. Taken together, these data provide valuable framework on Brucella induced miRNA expression in RAW264.7 cells, and suggest that Brucella may establish chronic infection by regulating miRNA expression profile.
Keywords: Brucella, RAW264.7, microRNA, high-throughput sequencing, apoptosis, autophagy