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Objective:To investigate the effect of metformin on the microRNA (miRNA) expression and screen potential target with network pharmacology analysis in patients with type 2 diabetes.Methods:Fifteen patients with new diagnosed type 2 diabetes admitted to our hospital were selected, who received metformin during hospitalization and after discharge. The expression of serum matrix metalloproteinase (MMP)-9, transforming growth factor (TGF)-β1, and myocardial fibrosis related miRNAs were compared before and 6 month after metformin treatment. In addition, gene ontology (GO) and KEGG pathway enrichment analysis were applied to analyze differential expression miRNAs showing statistical significance. Meanwhile, the network figure was established to reflect the target gene messenger RNA (mRNA) corresponding to differentially expressed miRNA.Results:Compared with pre-medication, the serum level of MMP-9 was significantly decreased after treatment ( P<0.05). Besides, the expression of homo sapiens microRNA (hsa-miR)29a-3p, hsa-miR133a-5p, hsa-miR21-5p, hsa-miR30c-5p, and hsa-miR1-3p in patients with type 2 diabetes were dramatically down-regulated by metformin ( P<0.05 or P<0.01). Results of GO analysis and KEGG pathway enrichment analysis showed that differentially expressed miRNAs were mainly concentrated in endoplasmic reticulum lumen, synapse, basement membrane and other cell components. The molecular functions such as Rho GTPase binding and participation in extracellular matrix structural constituent were exerted through biological processes such as collagen catabolic process, regulation of short-term neuronal synaptic plasticity, and axon extension, which were mainly enriched in advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway in diabetic complications, tumor necrosis factor (TNF) signaling pathway, and Wnt signaling pathway, etc. The outcome of miRNA-mRNA network analysis demonstrated that there were 230 target genes mRNAs corresponding to differentially expressed miRNA. Conclusion:Metformin could play its role in the treatment of type 2 diabetes by down regulating the expression of miRNA, participating in the transduction of related cellular signaling pathways, regulating chromatin, nucleic acid binding, and enzyme activities.
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@#Objective: To explore the genes that may be regulated by cell division cycle 25A (CDC25A) with gene chip technology, and to elucidate and verify that CDC25A has a regulatory effect on the expression of liver cancer related genes. Methods: CDC25A expression in human liver cancer HepG2 cells was silenced by siRNA interference technology and a nude mouse xenograft model of liver cancer was successfully constructed in our previous research. Affymetrix human gene expression profiling microarray was used to further screen differentially expressed genes (DEGs) after silencing CDC25A in liver cancer xenografts, and GO analysis and KEGG analysis were performed. Some of the DEGs were verified by qPCR. Results: The chip screened 188 DEGs in liver cancer xenograft tissues after CDC25A silence, including 78 up-regulated genes and 110 down-regulated genes. These DEGs mainly involved in cell proliferation, apoptosis, protein complex binding, extracellular space, etc., and associated with the changes in pathways such as focal adhesions and extracellular matrix (ECM) receptor interactions. qPCR showed that the expression of HIPK2 mRNA was up-regulated and the mRNA expressions of (microfibrillar-associated protein 5(MFAP5) and cyclin D1 (CCND1) were down-regulated, which were consistent with the results of microarray detection. Conclusion: Using human gene expression profiling chip, the DEGs in liver cancer xenograft tissues in nude mice after silencing CDC25Awere successfully screened, providing effective clues for exploring the effect of CDC25Aon the growth of liver cancer.
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Objective: To identify the changes of gene expression profiles and metastasis ability of rat hepatoma cell line McA-RH7777 after sublethal irradiation, and to explore the underlying molecular mechanism. Methods: Rat hepatoma McA-RH7777 cells were received single exposure of 6 Gy X-ray. Then, the remaining McA-RH7777 cells were continuously passaged and named as McARH7777- 6Gy. The gene expression profiles of McA-RH7777 and McA-RH7777-6Gy cells were detected by gene microarray and compared. Subsequent bioinformatic analysis of the genes with significant changes in expression levels were performed by DAVID software (including GO-analysis and KEGG-analysis). The mRNA expression levels of tissue inhibitor of metalloproteinase 2 (TIMP2), SMAD family member 2 (SMAD2) and MET proto-oncogene in McA-RH7777 and McA-RH7777-6Gy cells were detected by real-time fluorescent quantitative PCR to verify the difference in gene expression profiles of the two cells. The migration and invasion abilities of McA-RH7777 and McA-RH7777-6Gy cells were detected by scratch wound healing assay and Transwell chamber assay, respectively. Results: The gene microarray showed that the expressions of a series of tumor-related genes were changed in McA-RH7777-6Gy cells as compared with wild-type McA-RH7777 cells. Real-time fluorescent quantitative PCR showed that the relative expression levels of TIMP 2, SMAD 2 and MET in McA-RH7777-6Gy cells were raised by 13.000, 14.516 and 6.384 times as compared with McA-RH7777 cells, respectively. GO analysis showed that the remarkably changed genes mainly located in tumor metastasis-associated biological processes, such as Ras protein signal transduction, cell cycle arrest, cell migration, cell adhesion, negative regulation of apoptotic process, positive regulation of cell proliferation, positive regulation of epithelial to mesenchymal transition and positive regulation of MAP kinase activity. KEGG analysis showed these genes mainly involved in proteoglycans signaling pathway, phosphoinositide-3 kinase (PI3K)-protein kinase B (PKB, Akt) signaling pathway, viral carcinogenesis pathway, FoxO signaling pathway, Rap1 signaling pathway, Hippo signaling pathway, Ras signaling pathway, etc. The scratch healing and Transwell chamber tests showed that the migration and invasion abilities of McA-RH7777-6Gy cells were significantly enhanced as compared with McA-RH7777 cells (both P < 0.001). Conclusion: After sublethal radiation, the migration and invasion abilities of hepatoma cells are significantly enhanced, which may be related to the changes of gene expressions in metastasis-associated pathways.
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Background and purpose:Inlfammatory bowel diseases (IBD) are a group of chronic intestinal diseases, including ulcerative colitis (UC) and Crohn’s disease (CD). This study identified differentially expressed miRNAs in UC, CD and colitis-associated colorectal cancers (CAC) to explore their potential as novel molecular biomarkers. Methods:Tissue samples were taken from 13 UC patients, 3 CD patients, 12 CAC patients, and 8 age-and gender-matched healthy controls. The miRNA expressions were detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) assay. Known targets of deregulated miRNAs were utilized using miRWalk 2.0 database, and subsequent bioinformatics analysis of these target genes was performed by DAVID software (GO-analysis, KEGG-analysis and BIOCARTA-analysis). Results:The data showed that miR-146a, miR-27a, miR-29a, miR-20a and miR-21 were upregulated in UC, CD and CAC tissues compared with normal control. Moreover, the target genes of these miRNAs were enriched in several key signal transduction pathways including cancer-related pathway and immu-nity-associated pathway. Conclusion:miR-146a, miR-27a, miR-29a, miR-20a and miR-21 may play important roles in the switching from IBD to CAC.