Comparative study of joint bioinformatics analysis of underlying potential of ‘neurimmiR’, miR-212-3P/miR-132-3P, being involved in epilepsy and its emerging role in human cancer

Considering the critical roles of miR-132/212 participated in central nervous system, many researches started to explored the contributions of miR-132/212 to epilepsy and achieve something worthwhile. Further illuminates all the genes targeted by miR-132/212 may be a valuable means for us to completely understand the working mechanism playing in epilepsy, by which it can influence diverse biological process. This study attempts to establish macrocontrol regulation system and knowledge that miR-212-3p/132-3p effected the epilepsy, for this literature search, miRbase, Vienna RNAfold webserver, Human miRNA tissue atlas, DIANA-TarBase, miRtarbase, STRING, TargetScanhuman, Cytoscape plugin ClueGO + Cluepedia+STRING, DAVID Bioinformatics Resources, Starbase, GeneCards suite and GEO database are comprehensive employed, miR-132-3p/212-3p and its target gene were found have highly expressed in brain and lots of molecular function and metabolic pathways associated with epilepsy may be intervened by it. Meanwhile, the emerging role of miR-132-3p/212-3p being involved in human cancer also been analyzed by several webtools for TCGA data integrative analysis, most remarkably and well worth exploring in our research conclusion that showed miR-132-3p/212-3p may be the core molecular underlying tumor-induced epileptogenesis.


INTRODUCTION
Epilepsy is a common, serious neurological disorder characterized by recurring seizures due to abnormal neuronal excitability and which do latent harm to human health. However, the pathogenic mechanism resulted in epilepsy and resistance to currently available antiepileptic drugs (AEDs) remains poorly understood. Recently, accumulating evidence has found microRNAs (miRNAs) play pivotal modulators in pathogenesis and potential treatment for epilepsy [1]. In pathogenesis, miRNAs is thought to associate with large-scale changes to expression of gene modulating neuronal microstructure, cell death, neurotransmitter signaling, ion channels, gliosis, their rodent orthologues, share the same primary transcript to discern genes mediated by the cAMP-response element www.impactjournals.com/oncotarget/ Oncotarget, 2017, Vol. 8, (No. 25), pp: 40668-40682 Research Paper www.impactjournals.com/oncotarget binding (CREB) protein transcription factor which is important for neuronal development and function [6]. caused by neuronal stimulation, and the expression of miRmaturation, morphogenesis and function of neurons and whose dysregulation has more to do with a large amount of neurodegenerative disorders, such as tauopathies, schizophrenia, Alzheimer's disease, Huntington's disease, autism and the theme of this article, epilepsy. (Discussed in neuronal functions is the most studied, more evidences point towards an involvement of these miRNAs in human is associated with poor prognosis of colorectal cancer [11]. In most cases, miRNAs performed its regulating function in virtue of its target gene by a complementary manner, in which miRNAs guide RNA-induced silencing complex (RISC) to miRNAs response elements on target transcripts and usually lead to degradation or translational they may therefore target the same mRNAs and then involved in translational inhibition of these target genes [7]. In this maner, both miRs can regulate plenty of distinct gene target, and aid in the coordinate regulation of members of understand thoroughly how they are involve in pathological In the post-genomic era, a large amount of longfor the life sciences provide us means to systematically So, bioinformatics analyses were used in this study to and clinical database to explore the novel and worthwhile studying direction for epilepsy research. In the process of will often inadvertently be excavated out.

RESULTS
Mature sequence hsa-miR-132-3p and has-miR-212-3p may have a predominant role in nervous tissues used name in the vast majority of researches, but two respectively) originate from opposite arms of the same pre-miRNA have been found in human genome, so eventually, although there is also a small study miRof both miRs. Knowing the expression and distribution of miRs in different tissues is essential for understanding normal and disease development of respective tissue.
highly expressed in myocardia and muscle, and are well which indicated these miRs play a pivotal role in brain tissue development and impaired expression of them may result in disorders of the nervous system. ( Figure 1C) Experimentally validated target genes of miR-212-3p/miR-132-3p have highly expression in brain Before comprehensive target gene analysis, it's experimentally validated miRNA:gene interactions.

Computational analysis result of miR-132-3p and miR-212-3p-regulated biomolecular network
The occurrence of any disease involved some physiological and biochemical processes, such as biological process, molecular function, cellular component and metabolic pathways, these biological mechanism enrichment can provide www.impactjournals.com/oncotarget clues and references for further research, so Gene Ontology enrichment analysis of these validated genes was performed. The result of KEGG pathway revealed that these genes might as cell cycle, foxO signaling pathway, TGF-beta signaling pathway, MAPK signaling pathway, neurotrophin signaling pathway and several pathway about cancers. Biological process such as transcription, cell cycle, positive regulation of protein export from nucleus, synapse assembly, axon extension, brain development, axon regeneration are statistically enriched. Among these, transcriptions are primary regulation process "negative regulation of transcription from RNA polymerase II

The biomolecular information of remaining genes targeted by miR-132-3p and miR-212-3p
Meanwhile, all predicated targets but not been experimentally validated genes may have potential research values in recent years, so Targetscan, a target predicting programs with higher sensitivity and precision than other [17], was employed to

The emerging role of miR-132 and miR-212 participate in epilepsy
All target genes and whose biological process had been enumerate above, in order to better understand the can be rapidly induced by activation of neurons in vivo in both rats and patients with temporal lobe epilepsy  Combinate all target genes of all database and remove duplicates The genes may be related to biomolecular pathways of epilepsy The MalaCards human disease database integrates both specialized and general disease lists and can provide (cellular component, biological process and molecular epilepsy (Supplementary Table 5). Benzodiazepine Pathway, Pharmacodynamics and locomotion are the top 1 pathways and GO terms (Biological processes) related to epilepsy, respectively (more details can be found in MalaCards). In this article, we want to assure whether genes may be targeted by both miRs. Among these genes, only IER3IP1 have been validated, SLC6A1, SCN1A, For conforming the fact that these genes did were implicated in epilepsy, STRING analysis was carried out epilepsy is the directly target genes in theory, we found several of them can be indirectly mediated by both miRs through protein-protein interactions and common biomolecular pathway (data not shown).

miR-132-3p and miR-212-3p may play a critical role in human cancer
Gene ontology and pathway enrichment analysis about validated and remaining target genes indicated miRcancer. For experimental validated genes and predicted genes statistically enriched pathways are directly correlated with prostate cancer, bladder cancer, proteoglycans in cancer, Glioma, endometrial cancer, colorectal cancer and so forth.   showed 5p also have a high concordance of brain tissueof miRNA and its target genes across human tissues, the result provide a sound basis for its involvement in Because miRNAs execute functions through target genes and corresponding biological processes that these target genes and enrichment of gene-GO term and bioenriched pathways have been previously implicated in epileptic seizure or related activities, such as decreased interaction between FoxO3a and Akt correlates with neurotrophin signaling pathway and other disorders such as temporal lobe epilepsy enrichment analysis of GO, which is similar to the fact that microRNAs may play important role in transcriptional analysis of target gene, the same analysis was carried on remaining predicted targets, Axon guidance in these genes, previous study have found hippocampal axon guidance can be regulated by nor-1 and involve in mTOR signaling pathway can sever as a new therapeutic strategy in epilepsy and epileptogenesis, all which absented in validated target genes may be provide a valuable reference and guide for Since so many biological mechanism are related to epilepsy, we are interested in the current research Pubmed search found 15 papers about these neurimmiRs PI3K and MAPK pathways, which is accorded with the above analysis. Comprehensive comparison, we feel that the study on involvement of both miRs in epilepsy is still less, there is still a large research space. Certainly, epilepsy is also belong to CNS disorder and results from a variety of CNS insults, theoretically, all disease caused by dysregulated expression of miRseizures.
In addition to Pubmed search, the MalaCards been used widely was also used for further analysis of as the main research object, as data are accumulated and analysis continues, the result demonstrated miRin human cancer, For experimental validated genes and cancer directly, respectively. Such as pathways in cancer, MicroRNAs in cancer and Proteoglycans in cancer, which is consistent with previous literatures about the emerging Moreover, both miRs may perform tumour-promoting and tumour-suppressing dual functions depending on cancer types, which is consistent with what been previously reported [7]. Our group also found both miRs present All unveil the complexity of both miRs involvement in tumourigenesis.
with selected core validated target genes, the double role of miRs functioned in caner been further validated. What itself, all indicated miRs may target single gene to interfere the process of cancer and epilepsy. Certainly, as epilepsyassociated genome is rapidly increase, the tally of genes www.impactjournals.com/oncotarget

Analysis of biological networks for validated target gene of miR-212-3P/miR-132-3P
All enrichment analysis of gene-GO term and bioplugin ClueGO + Cluepedia+STRING app and DAVID of protein-protein interactions. All gene tissue expressions in human tissue were assessed by DAVID Bioinformatics Resources 6.8. Besides, oriented to this article focused on epilepsy, literature search (PubMed) was carried out to analyze

Analysis of all the target gene of miR-212-3P/ miR-132-3P
TargetScanhuman7.1 was used to predicts biological (refer to the gene that have ruled out the experimentally validated target gene from all predicted target gene) performed enrichment analysis of gene-GO term, biopathways as described above.

Analysis of genes and related bio-pathways may be related to epilepsy using GEO data and MalaCards
To screen the differential expression gene between patients with and without epilepsy, GEO super-series normalized expression data were download from NCBI and the bio-pathways of these differential expression genes was analyzed as above, Volcano Plot and dendrogram of different genes was analyzed through Gene-Cloud of Biotechnology Meanwhile, in order to include more genes associated with epilepsy supportedby literature, the data compiled from Function enrichment analysis is performed as above.