Target Gene Of miR-132 Research Articles

MiR-132 Inhibits Tumor Metastasis

It has been shown that some miRNAs are related with tumor invasion and metastasis directly or indirectly.Our aim is to find a specific miRNA which plays an essential role in tumor metastasis.Although it has been shown that miR-132 was associated with blood vessel growth,neural development and differentiation,and inflammation,relationship between miR-132 and tumor metastasis was not studied in any research.In order to identify the effect of miR-132 on tumor metastasis,the migration ability in vitro was detected on breast cancer cell line MDA-MB-231 cells transducted with miR-132(miR-132 group) and or mock miRNA(control group) by transwell assay.The results demonstrated significantly lower migration ratio in miR-132 case compared to that in control case,indicating inhibition effects on cancer migration of miR-132.To clarify the inhibition mechanism by which miR-132 inhibits cancer metastasis,target genes of miR-132 were screened and identified.They are CHIP(STUB1),G3BP1 and G3BP2.The expression levels of these 3 genes in MCF7 cells(metastasis cell line) and MDA-MB-231 cells with or without transduction of miR-132 or mock miRNA were detected by PCR and Real-time PCR.Two key genes,G3BP1/G3BP2,were founded to be involved in the regulation of miR-132 to tumor metastasis,demonstrating that miR-132 could silence G3BP1/G3BP2,which resulted in the suppression of tumor metastasis.Our research suggests that miR-132 may be an important potential target used for inhibition of cancer metastasis and clinical therapy of cancer,and shed light on the suppression mechanisms of miR-132.

MicroRNA-132 dysregulation in schizophrenia has implications for both neurodevelopment and adult brain function

Schizophrenia is characterized by affective, cognitive, neuromorphological, and molecular abnormalities that may have a neurodevelopmental origin. MicroRNAs (miRNAs) are small noncoding RNA sequences critical to neurodevelopment and adult neuronal processes by coordinating the activity of multiple genes within biological networks. We examined the expression of 854 miRNAs in prefrontal cortical tissue from 100 control, schizophrenic, and bipolar subjects. The cyclic AMP-responsive element binding- and NMDA-regulated microRNA miR-132 was significantly down-regulated in both the schizophrenic discovery cohort and a second, independent set of schizophrenic subjects. Analysis of miR-132 target gene expression in schizophrenia gene-expression microarrays identified 26 genes up-regulated in schizophrenia subjects. Consistent with NMDA-mediated hypofunction observed in schizophrenic subjects, administration of an NMDA antagonist to adult mice results in miR-132 down-regulation in the prefrontal cortex. Furthermore, miR-132 expression in the murine prefrontal cortex exhibits significant developmental regulation and overlaps with critical neurodevelopmental processes during adolescence. Adult prefrontal expression of miR-132 can be down-regulated by pharmacologic inhibition of NMDA receptor signaling during a brief postnatal period. Several key genes, including DNMT3A, GATA2, and DPYSL3, are regulated by miR-132 and exhibited altered expression either during normal neurodevelopment or in tissue from adult schizophrenic subjects. Our data suggest miR-132 dysregulation and subsequent abnormal expression of miR-132 target genes contribute to the neurodevelopmental and neuromorphological pathologies present in schizophrenia.

Regulation of miR-132 on Target Gene FOXA1 in Esophageal Carcinoma Cell Line KYSE150

Objective To construct an eukaryotic expression vector of miR-132 and FOXA1,to verify the regulation effect of miR-132 on the target gene FOXA1 in human esophageal carcinoma cell KYSE150.Methods According to sequence of the mature miR-132 with its flank sequences,PCR primers were designed and miR-132 precursor sequence was amplified from genomic DNA of EC9706 cell.PCR product was cloned into pMD18-T Simple vector and then subcloned into pCDNA3.1(+) between BamHⅠand EcoRⅠ.Human esophageal carcinoma cell line KYSE150 was transfected by pCDNA3.1(+) and pCDNA3.1-miR-132,miR-132 were detected by real-time PCR.FOXA1 was predicted by using bioinformatics.The 3′UTR of candidate target gene FOXA1 was cloned into the downstream of pMIR vector,mutation expression vector pMIR-FOXA1-Mut was also constructed.Co-transfection of both vectors were performed in KYSE150 cells and dual-luciferase reporter assay was analyzed.Western blot was used to detect the expression of FOXA1 protein in KYSE150 cell line transfected with pCDNA3.1(+)-miR-132 and pCDNA3.1(+) respectively.Results The miR-132 expression vector has been constructed successfully and it can effectively express mature miR-132 in esophageal carcinoma cell KYSE150.FOXA1 was selected as a candidate of target genes which has 7 base pairs completely matched to miR-132 in the 3′UTR seed region.Recombined vector pMIR-FOXA1 fused FOXA1 3′UTR and mutation expression vector pMIR-FOXA1-Mut were constructed.Dual-luciferase reporter assay showed that the luciferase activity decreased in the group co-transfected with pCDNA3.1(+)-miR-132 and pMIR-FOXA1.Overexpression of exogenous miR-132 in KYSE150 cell line can significantly suppress the expression of FOXA1 protein.Conclusion miR-132 can suppress FOXA1 gene expression at the post-transcriptional level by targeting to the specific sequence of FOXA1 gene 3′UTR,FOXA1 is a target gene of miR-132.