| Objectives To screen and verify the differentially expressed micro RNA in plasma in patients infected with non-tuberculosis mycobacteria(NTM) when compared with patients infected with mycobacterium tuberculosis(MTB), and then to predict its target genes and analyses its biological function.Methods 1 The choice of objectives: A total of 56 TB patients infected with NTM(19cases) and MTB(37 cases) were were collected from The Fourth Hospital in Tangshan and The Fifth Hospital in Shijiazhuang from October 2013 to October 2014. After achieving the patients’ agreement, we extract their plasma and then investigate their general conditions. six objectives were chosed from each group respectively were used for Genechip screening,the other objectives were used for the Quantitative reverse transcription polymerase chain reaction(RT-q PCR). 2 Gene Chip mi RNA Arrays were used to screen the differentially expressed mi RNAs in NTM groups when compared with MTB group, t test were used to analysis the mi RNA expression level, P<0.05 means statistically difference. RT-q PCR method was used to test the reliability of the Gene Chip results, 2-△△Ctwere used to measure the relative expression of mi RNA in NTM group and MTB group, 2-△△Ct>1 meas up-regulation and 2-△△ Ct<1 means down-regulation. 3The Mi Rwalk, Targetscan and Mi Randa databases were used at the same time to predict the target genes. The Database for Annotation, Visualization, and Integrated Discovery was used to analyze the enriched gene ontology pathway and gene function. 4Statistical analysis method: SPSS17.0 was used to analysis the general situation, t test was used to analysis the age information,2? test was used to analysis the classification information, P<0.05 means statistically difference.Results 1 The screening and predication of the differentical expression mi RNA:(1)According to the result of Gene Chip mi RNA Arrays and the previous studies reports,8 up-regulated mi RNAs and 3 down-regulated mi RNAs were validated by the Target Scan, mi Rwalk and mi Randa databases simultaneously. The up-regulated mi RNAs were hsa-mi R-26 a, hsa-mi R-24, hsa-mi R-222, hsa-mi R-191, hsa-mi R-155, hsa-mi R-126, hsami R-122, hsa-let-7b, whereas the down-regulated mi RNA were mi R-767, mi R-1283, and mi R-1281.(2)The results of the RT-q PCR: in the study, six significance differentially expressed mi RNA(has-let-7, has-mi R-155, has-mi R-26 a, has-mi R-1281, has-mi R-1283,has-mi R-767) were chosen to verify the results of Gene Chip, and the results of the RTq PCR were same to the Genechip results. 2 The results of target gene prediction and biological function:(1)A total of 23 target genes were regulated by the up-regulated mi RNAs; 59 target genes were regulated by the down-regulation mi RNAs, According to the analysis, these target genes mainly regulate the following two biological functions: on the one hand, they mainly regulate the innate immune response and influence phagocytosis of germs of the phagocytosis lysosome to influence the inflammatory response; on the other hand, they also regulate the cell apoptosis by influencing some related signal paths.(1)The results of Gene Ontology: in this study, the 8 up-regulated mi RNA mainly participate in 42 signal paths, they were mainly influence the inflammatory response and the cell cycle by regulating protein kinase and gluconeogenesis process. The 3 down-regulated mainly participate in 8 signal pathes.They were mainly influence the cell apoptosis by regulating some metabolic pathways and signal transduction pathways.Conclusios 1 There were 8 up-regulated mi RNA and 3 down regulated mi RNA in the NTM group compared with the MTB group. 2 In this study, the difference expression mi RNA mainly regulate the inflammatory reactionthe, cell apoptosis and the cell metastasis... |
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