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@#Metastasis-associated lung adenocarcinoma transcript 1(MALAT1)is one of the first identified LncRNA associated with human diseases. Unlike most members of the LncRNA family, MALAT1 is found in almost all human tissues and expressed at a relatively high level. At present, MALAT1 is known to play a vital role in the pathophysiological process of many diseases such as tumors, cardiovascular diseases, and nervous system diseases. In recent years, studies have found that MALAT1 may be involved in many ocular diseases(such as diabetic retinopathy, cataracts, glaucoma, retinoblastoma, neonatal retinopathy, <i>etc</i>.)play an important role in the pathological development process, and it is expected to become an effective target for the diagnosis and treatment of eye diseases. This article summarizes the research progress of eye diseases in which MALAT1 has participated in recent years.
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Objective @# To observe the clinical significance of miR-135b-5p in oral squamous cell carcinoma (OSCC) tissues and to conduct a bioinformatics analysis of its predicted target genes.@*Methods @#The expression levels of miR-135b-5p in OSCC tissues and adjacent normal tissues were compared using data from TCGA and GEO databases, and the correlations of miR-135b-5p expression level with clinicopathologic characteristics were analyzed. Fresh tissues were collected in the clinic, and the expression of miR-135b-5p was verified by quantitative real-time PCR. The target genes with enriched pathways were analyzed by using bioinformatics methods. A protein-protein interaction network was constructed to screen hub genes.@*Results @#The expression levels of miR-135b-5p were significantly upregulated in OSCC tissues compared to adjacent normal tissues (P < 0.001) and had a good diagnostic capability (AUC=0.960, P < 0.001). The expression level of miR-135b-5p was positively correlated with histopathological grading (P=0.011). Enrichment analyses revealed that the target genes of miR-135b-5p were significantly associated with tumor-related signaling pathways, such as the calcium signaling pathway, the cGMP-PKG signaling pathway and the cAMP signaling pathway. Ten core target genes were obtained by screening: DLG2, ANK3, ERBB4, SCN2B, NBEA, GABRB2, ATP2B2, SNTA1, CACNA1D, and SPTBN4.@*Conclusion@#miR-135b-5p may act as an oncogene miRNA in OSCC and has the potential value of acting as a diagnostic biomarker and therapeutic target for OSCC.
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Objective:To investigate the differential expression profile of circular RNA (circRNA) in high glucose-cultured human retinal vascular endothelial cells (HRVECs).Methods:HRVECs were divided into high glucose group, normal control group and hypertonic control group, and were cultured in 25 mmol/L glucose medium, 5.5 mmol/L glucose medium and 19.5 mmol/L mannitol+ 5.5 mmol/L glucose medium for 24 hours accordingly.The differentially expressed circRNA molecules between high glucose group and normal control group were screened by circRNA microarray analysis.The expression of the most significant differentially expressed circRNAs in different groups was verified by real-time quantitative PCR.The possible microRNA (miRNA) targets were analyzed through the Circular RNA Interactome database.Results:It was found that 448 circRNAs were differentially expressed (FC≥1.5 or FC≤0.67, P<0.05) in high glucose-cultured HRVECs, among which 182 were up-regulated and 266 were down-regulated.The top 3 significantly up-regulated circRNAs were hsa_circ_0002938, hsa_circ_0008036, and hsa_circ_0001946, and the top 3 significantly down-regulated circRNAs were hsa_circ_0035277, hsa_circ_0008344, and hsa_circ_0001874.Compared with normal control group and hypertonic control group, the relative expressions of top 3 up-regulated circRNAs were significantly enhanced and the relative expressions of top 3 down-regulated circRNAs were significantly reduced in high glucose group, showing statistically significant differences (all at P<0.05).No significant difference was found in the differentially expressed circRNAs between normal control group and hypertonic control group (all at P>0.05). Conclusions:CircRNAs are differentially expressed in high glucose-cultured HRVECs, and the differentially expressed circRNAs may be involved in the regulatory mechanism of diabetic retinopathy.
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Objective:To analyze differentially expressed microRNAs (miRNAs) and target genes in human respiratory epithelial cells (16HBE) after enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) infection using high-throughput sequencing.Methods:TargetScan and miRDB databases were used to predict the target genes of miRNAs that were both up-regulated or down-regulated after EV71 and CVA16 infection. The genes that were both up-regulated and down-regulated were screened out. GO and pathway analysis of the target genes were conducted to screen immune-related target genes and their corresponding miRNAs. The target genes and their corresponding miRNAs that were up-regulated or down-regulated in both immune-related GO and pathway were further screened. Some miRNAs and their target genes were selected for qRT-PCR verification.Results:There were 598 target genes of up-regulated miRNAs and 1 311 target genes of down-regulated miRNAs and 62 target genes that might be up-regulated or down-regulated simultaneously were screened out. The number of up-regulated target genes involved in immune-related GO and pathway were 17 and 13, respectively, and the number of corresponding miRNAs were 15 and 17, respectively. There were 58 and 47 down-regulated target genes involved in immune-related GO and pathway, respectively, and the number of corresponding miRNAs were 30 and 42, respectively. Three up-regulated target genes were involved in both immune-related GO and pathway and regulated by four miRNAs. Nine down-regulated target genes were involved in both immune-related GO and pathway and regulated by 13 miRNAs.Conclusions:This study was conducive to elucidate the host-pathogen interaction after EV71 and CVA16 infection, and provided reference for studying the pathogenesis of hand, foot and mouth disease.
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OBJECTIVES@#White matter hyperintensity (WMH) is an important factor leading to cognitive impairment, and the mechanism has not been clarified. In recent years, studies have found that circular RNA (circRNA) has differential expression in cerebrovascular diseases. This study aims to analyze the expression profile of circRNA in peripheral blood mononuclear cell (PBMC) of patients with WMH with cognitive impairment, to screen the differentially expressed circRNA, and to explore the possible role of circRNA in WMH with cognitive impairment.@*METHODS@#CircRNA microarray was used to detect the circRNA expression profile of PBMC in patients with WMH with cognitive impairment, and in patients with WMH without cognitive impairment as well as in normal controls (3 cases each, male to female ratio of 2꞉1). The differentially expressed circRNA in patients with WMH with cognitive impairment was screened. The screening criteria for differentially expressed circRNA was fold change (FC) ≥2.0 (|log@*RESULTS@#Compared with the control group, there were 5 significantly up-regulated circRNA and 3 down-regulated circRNA in the WMH with cognitive impairment group; 8 circRNA were significantly up-regulated and 2 were down-regulated in the WMH without cognitive impairment group. When compared with the WMH with cognitive impairment group, no co-differentially expressed circRNA was found in WMH without cognitive impairment group and control group. Compared with the control group, the expression of hsa_circ_0092222 was up-regulated and the expressions of hsa_circ_0000662 and hsa_circ_0083773 were down-regulated in the WMH with cognitive impairment group and the WMH without cognitive impairment group, and there was no significant difference between the 2 groups (all @*CONCLUSIONS@#The circRNA expression profile of patients with WMH is changed significantly. The differentially expressed circRNA may be the cause of WMH; Hsa_circ_0092222, hsa_circ_0000662, and hsa_circ_0083773 may regulate the expression of target genes by targeting adsorption of the target miRNA, leading to brain white matter damage through Janus kinase 2 (JAK2)/signal transducers and activators of transcription (STAT3) signal pathway and Wnt signal pathway.There is no significant difference in circRNA expression profile between WMH with or without cognitive impairment. Cognitive impairment in patients with WMH may have other reasons.
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Female , Humans , Male , Cognitive Dysfunction/genetics , Leukocytes, Mononuclear , MicroRNAs , RNA/genetics , RNA, Circular , Software , White MatterABSTRACT
Objective:To explore the genes and molecular markers related to the sensitivity to concurrent chemoradiotherapy in patients with locally advanced esophageal squamous cell carcinoma.Methods:The peripheral blood sample of 31 patients with locally advanced esophageal squamous cell carcinoma receiving radical concurrent chemoradiotherapy was collected and the plasma circulating free DNA (cf-DNA) was extracted before treatment. The target gene capture sequencing technology based on NovaseQ6000 high-throughput sequencing platform was employed to detect the changes of target genes and tumor mutation burden (TMB). According to the short-term efficacy of chemoradiotherapy, all patients were divided into the chemoradiotherapy-sensitive group (CR+ PR) and chemoradiotherapy-resistant group (SD+ PD). Bioinformatics and clinical data were adopted to analyze the differences of gene mutation and TMB between two groups.Results:In the sequencing data of 31 patients, the tumor-related genes with a mutation frequency above 10% were Tp53, Notch1, BRAF, FGFR4, CDKN2A, ATRX and Axin2, which were almost equally distributed between the CR+ PR and SD+ PD groups. High-frequency mutant genes were associated with 7 signaling pathways, mainly involved in the RTK/RAS signaling pathways. The TMB value in the CR+ PR group was higher than that in the SD+ PD group ( P=0.04), however, the mutation rate of GXYLT1 and KRT18 genes in the SD+ PD group was higher than that in the CR+ PR group ( P<0.05). Conclusions:Tp53, Notch1 and CDKN2A may be the high-frequency mutant genes associated with the incidence and progression of esophageal squamous cell carcinoma. KRT18, GXYLT1 and TMB are closely correlated with the sensitivity to concurrent chemoradiotherapy of patients with locally advanced esophageal squamous cell carcinoma.
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【Objective】 To analyze the changes of microRNA (miRNA) expression profiles on day 1 and day 5 after storage with or without riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample), collected from voluntary donors, were split into two group after mixing and agitation. One was treated with riboflavin (final concentration 50 μmol/L) plus 6.24 J/mL UVB light(E group), and the other worked as a control group (C group) without any treatment. Both groups were subjected to agitated storage at (22±2) ℃ horizontally. The platelet concentrates were sampled on d1 and d5 (5mL) during storage, named as E1, E5, C1 and C5 groups, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between E and C groups were screened by using DEGseq and MA-plot analysis software, and GO function enrichment analysis and KEGG pathway enrichment analysis were further performed when the different expression between groups reached twofold and above. 【Results】 Compared with C1 group, 487 miRNAs with significantly different expression (P<0.01) were screened in E1 group, including 220 up-regulated miRNAs, such as miR-146a and let-7b, and 267 down regulated miRNAs, such as miR-7 and miR-1260. Compared with C5 group, 229 miRNAs with significantly different expression (P<0.01) were screened in E5, including 80 miRNAs with up-regulated expression, such as miR-423 and miR-378, and 149 down regulated miRNAs, such as miR-451 and miR-30.The target genes with differentially expressed miRNAs in E1 vs C1 groups and E5 vs C5 groups were similar in the numbers of enriched GO terms, including cell components, organelles, cell membrane and other cell structures, molecular functions such as adhesion, catalysis, molecular transformation, transportation, transcription factors and receptor activity, cell processing, metabolism, biological regulation, stress and other biological processes etc. Compared with E1 and C1 groups, E5 and C5 groups lacked of signal pathways related to environmental adaptation, translation and mucin synthesis, however, it increased inositol phosphate metabolism, phosphatidylinositol signaling system and chemokine signaling pathway. 【Conclusion】 The expression profiles of platelets miRNAs treated with VB2-PRT has changed significantly after storage for a period of time. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL induced by VB2-PRT.
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【Objective】 To investigate the changes of platelet microRNA (miRNA) expression profiles on d1 and d5 during storage with riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample) were collected from voluntary blood donors. After mixing and shaking, the samples was treated with riboflavin (final concentration 50 μmol/L) and 6.24J/mL UVB light for 8min, then split into two aliquots and agitated stored at (22±2) ℃. The concentrates were sampled (5mL) on d1 and d5, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between the two groups (at different storage periods) were screened by DEGseq and MA-plot analysis software. The miRNAs, reached more than 2 times different expression between groups, were considered significant different(P<0.01). The miRanda and TargetScan softwares were used to predict the target genes. Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed on the target genes of significant differentially expressed miRNAs. The expression of miRNAs was verified by real-time fluorescence quantitative PCR (qRT-PCR). 【Results】 miRNA expression profile: compared with d1 platelets, there were 590 miRNAs with significantly different expression (P< 0.01) in d5 group, including 255 up-regulated miRNAs (such as miR-99b, miR-7) and 335 down regulated miRNAs (such as miR-451a, miR-19b). Among the 272 known miRNAs, 112 were up-regulated and 160 were down regulated. There were 318 new miRNAs sequences. The enriched GO terms of target genes of differentially expressed miRNAs in d5 and d1 groups included cell components, organelles, cell membranes and other cellular structures, molecular functions such as adhesion, catalysis, molecular conversion, transportation, transcription factor and receptor activity, and biological processes such as cell processing, metabolism, biological regulation and stress. The corresponding pathways in the top 10 of KEGG enrichment were mainly secretion, glucose metabolism, signal transduction, membrane transport, translation, environmental adaptation and other signal pathways. The six randomly selected differentially expressed miRNAs verified by qRT-PCR was consistent with those of DNB sequencing. 【Conclusion】 The expression profiles of platelets miRNAs has changed significantly between d1- and d5-storage under VB2-PRT treatment. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL underVB2-PRT treatment.
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Objective@#To explore the genetic basis of a child with idiopathic mental retardation.@*Methods@#Clinical data and peripheral blood sample of the child were collected. Genomic DNA was extracted and subjected to copy number analysis using single nucleotide polymrophism array comparative genome hybridization (SNP-aCGH) and targeted capture and next generation sequencing (NGS).@*Results@#No microdeletion/microduplication were detected by SNP-aCGH. NGS has detected homozygous c. 722delA(p.Asp241fs) variant of the LISN1 gene, which is known to underlie autosomal recessive mental retardation - 27 (MRT 27). Both parents are carriers of the variant, conforming to the autosomal recessive inheritance.@*Conclusion@#A novel pathogenic variant of the LINS1 gene has been identified, which probably underlies the MRT 27 in the patient.
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ObjectivecircRNAs play an important role in tumor development, but the relationship between circRNAs and hepatocellular carcinoma remains to be further explored. The present study aimed to bioinformatically analyze the target gene of microRNA-1. Another aim was to screen circRNAs that are associated with target genes and differentially expressed in hepatocellular carcinoma cells, as well as provide theoretical basis for clinical screening of molecular markers and targeted therapies related to hepatocellular carcinoma.MethodsThe miRNA related database used for the prediction of microRNA-1 target genes, and the bioinformatic analysis of the target genes of microRNA-1 involved functional enrichment analysis and signal transduction pathway enrichment. Then, the circRNAs, which are related to the downstream target genes of microRNA-1, are screened through the circRNA database.ResultsThe number of microRNA-1 target genes was 230 in miRNA related database. Through GO analysis, it was found that the target genes of microRNA-1 had a strong tendency in regulation, and were mainly enriched in three aspects: biological function, biological process and cell localization.The target genes of microRNA-1 are involved in the function of proteins, regulation of biosynthesis, cofactor binding, enzyme regulation and other biological processes. Predicted target genes of miRNA-1 were significantly enriched in cancer signaling pathways, hepatitis B occurrence, endocytosis and splicing pathways. Further, 21 circRNAs related to the target gene of microRNA-1 were found in three circRNA databases, wherein hsa_circ_0004651 was highly expressed in hepatocellular carcinoma cell line HepG2 and its pavent gene was hnRNPD.ConclusionMicroRNA-1 influence the occurrence of hepatocellular carcinoma development through the regulation of protein and enzyme. Hsa_circ_0004651 may affect the development of hepatocellular carcinoma with microRNA-1 and its parental gene hnRNPD.
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Objective:To investigate the targets and possible mechanism of Didangtang in the treatment of bladder cancer. Method:Based on multiple traditional Chinese medicine and disease databases, the network pharmacology was used to screen potential targets, analyze the biological functions of potential targets, and construct a network of "Chinese medicine-target-path-disease". Bioinformatics analysis was applied in population and gene databases, in order to explore the differential expressions of core targets in tissues, distribution in the population and the correlation with prognosis. The in vitro experiment was used to verify the biological function of Didangtang. The underlying mechanism of Didangtang on the candidate target was detected. Result:A total of 21 core target genes and 16 highly enriched pathways were screened out. A functional network of Didangtang was constructed systematically. At the same time, six targets, namely cadherin 1 (CDH1), CAMP responsive element binding protein 1 (CREB1), colony stimulating factor 2 (CSF2), AP-1 transcription factor (JUN), matrix metalloproteinase 2 (MMP2), and prostaglandin-endoperoxide synthase (PTGS2), were differentially expressed in bladder cancer tissues (P<0.05). Furthermore, JUN and MMP2 were also differentially distributed in population (P<0.05). At the same time, the expression level of JUN was correlated with the prognosis of patients with bladder cancer (P<0.05). The in vitro experiment revealed that Didangtang inhibited the proliferation of bladder cancer cells and decreased the expression of candidate target JUN (P<0.01). Conclusion:Didangtang has the characteristics of multiple targets and multiple pathways in treatment of bladder cancer. It is initially confirmed that Didangtang can affect the expression of target JUN and inhibit the proliferation of bladder cancer, which lays a good foundation for further studies on mechanism.
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@#To investigate the effect of miR-802 on insulin secretion by islet β cells and its mechanism, miR-802 was overexpressed or knocked down in primary islet cells and Min6 cells via transfecting miR-802 mimic and miR-802 inhibitor, respectively. The effect of miR-802 on insulin secretion was detected by ELISA. The target gene of miR-802 was confirmed by miRNA target gene database prediction, luciferase report and Western blot. The function recovery experiment was carried out to clarify the mechanism of miR-802 regulating β cell secretion of insulin. The results showed that overexpression of miR-802 in islet primary cells and Min6 cells inhibited insulin secretion. qPCR and Western blot showed that miR-802 inhibited insulin secretion by inhibiting the transcription and translation of the target gene, hepatocyte nuclear factor 1β(Hnf1B).
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@#Oral squamous cell carcinoma (OSCC) is the most common oral cancer. Previous studies have found significantly high miR-155 expression in OSCC. However, the mechanism by which miR-155 plays a role in OSCC oncogenesis is not yet clear. This article reviews the function of the relationship between miR-155 and tumors and the potential role of miR-155 in the development of OSCC. A literature review showed that mir-155, as a small carcinogenic RNA, can inhibit CDC73, BCL6, P27Kip1 and other target genes that play a role in cancer inhibition; promote the proliferation, migration and invasion of OSCC cells; and inhibit apoptosis. miR-155 can also be combined with biological factors (Epstein-Barr virus, human papillomavirus) to promote the development of OSCC.
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Objective • To detect the effects of miR-760 on proliferation, apoptosis, migration and invasion of human esophageal squamous cell carcinoma (ESCC) TE-10 cells and to analyse the underlying mechanism. Methods • The mRNA and protein expression levels of miR-760 and c-Myc in five ESCC cell lines (normal esophageal epithelial cells as control) and 14 pieces of ESCC tissue specimens (paracancerous tissues as control) were detected by using reverse transcription quantitative PCR (RT-qPCR) and Western blotting. TE-10 cells were transfected with miR-760 mimics, miR-760 inhibitor (miR-mimics/miR-inhibitor group) and corresponding negative controls (mimics-NC/inhibitor-NC group), in which the overexpression and inhibition efficiency of miR-760 and the expression of c-Myc were verified by RT-qPCR. The effect of miR-760 on the proliferation of TE-10 cells was assessed by CCK-8 and colony formation assay. Changes of cell cycle distribution and proportion of apoptotic cells were measured by flow cytometry. Expression levels of cell cycle-, apoptosis-, migration-, and invasion-associated proteins as well as c-Myc were analyzed by Western blotting. The targeting relationship between miR-760 and c-Myc was verified by using the dual luciferase reporter assay. Results • The expressions of miR-760 were down-regulated and the expressions of c-Myc mRNA and protein were up-regulated in five ESCC cell lines compared with those in the normal esophageal epithelial cells. In 14 cases of ESCC tissue specimens, the expressions of miR-760 were down-regulated but the expressions of c-Myc were up-regulated compared with those in the cancer-adjacent tissues. The proliferation ability of TE-10 cells in the miR-mimics group was markedly attenuated, and colony numbers were also decreased. Flow cytometry assay showed that the proportion of cells in G1 phase was notably augmented, and the proportion of apoptotic cells was also increased. The miR-mimics group cells had weaker migration and invasion potential compared with mimics-NC group. Western blotting analysis conformed that expression levels of cyclin D1, B cell lymphoma 2, matrix metalloproteinase 2 and vimentin were decreased, but expression levels of cleaved-caspase3, E-cadherin and β-catenin were elevated. The miR-inhibitor group showed opposite results compared with the miR-mimics group. The dual luciferase reporter assay validated the direct targeted binding of miR-760 to the 3'UTR of c-Myc. Conclusion • miR-760 can suppress proliferation, migration and invasion, and induce apoptosis of TE-10 cells by directly targeting c-Myc.
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Objective:To analyze and identify ginseng miRNA. Methods: The miRNA and target genes of Shizhu ginseng and Yuan ginseng were detected by the degradation sequencing technology; Functional annotation of Degradome genes was carried out using public databases of KEGG/NR/GO database; The expression of miRNA and target genes of Shizhu ginseng and Yuan ginseng was determinated by real time fluorescence quantitative PCR technique. Results: A total of 13 target genes of eight miRNA families were obtained; The target gene type of miRNA was mainly transcriptional factor, response factor, and signal transduction pathway by means of KEGG/NR/GO database analysis. The results of real time fluorescence quantitative PCR verification of aqc-miR-159, bdi-miR162, cpa-miR319, pgi-miR4376, smo-miR396 and its target genes were basically consistent with the expression of miRNA and target genes from the degradation group. Conclusion: The target genes of partial Panax ginseng miRNA is clarified, which lays the foundation for further study of the possible function of ginseng miRNA.
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Objective@#To find the differential expression profiles of circRNA in whole blood and predict its target genes in blood of patients with tuberculosis in Xinjiang, and explore the relationship between circRNA and the development of tuberculosis. @*Methods@#The circRNAs expression in peripheral blood from 3 pulmonary tuberculosis patients and 3 healthy individuals were tested by using circRNA microarray assay. The whole blood from 43 patients with tuberculosis, 40 healthy individuals and 43 patients with pneumonia were collected to verify the results by real-time quantitative PCR system. The possibility of differentially expressed circRNA target genes were predicted by circRNA target gene prediction database. @*Results@#In the results of microarray assay 835 circRNAs were found to be expressed differentially in whole blood between the tuberculosis patients and healthy controls of Xinjiang area, of which 249 circRNAs were up-regulated and 586 circRNAs were down-regulated in the patients. The expressions of four significantly different circRNA were verified by real time quantitative PCR and the results showed that hsa_circ_0008276, hsa_circ_0003452, hsa_circ_0001846 and hsa_circ_0090508 were down-regulated (P＜0.05), and hsa_circ_0090508 was the more specific than the other three circRNAs. The results of circRNA target genes prediction suggested that has-miR-1294, has-miR-604, has-miR-616, has-miR-663b and has-miR-486-3p may be the potential target genes of hsa_circ_0090508. @*Conclusion@#The differentially expressed circRNA hsa_circ_0090508 was significantly downregulated in the patients with tuberculosis and may affect the regulation mechanism of tuberculosis through target genes.
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Objective@#To find the differential expression profiles of circRNA in whole blood and predict its target genes in blood of patients with tuberculosis in Xinjiang, and explore the relationship between circRNA and the development of tuberculosis. @*Methods@#The circRNAs expression in peripheral blood from 3 pulmonary tuberculosis patients and 3 healthy individuals were tested by using circRNA microarray assay. The whole blood from 43 patients with tuberculosis, 40 healthy individuals and 43 patients with pneumonia were collected to verify the results by real-time quantitative PCR system. The possibility of differentially expressed circRNA target genes were predicted by circRNA target gene prediction database. @*Results@#In the results of microarray assay 835 circRNAs were found to be expressed differentially in whole blood between the tuberculosis patients and healthy controls of Xinjiang area, of which 249 circRNAs were up-regulated and 586 circRNAs were down-regulated in the patients. The expressions of four significantly different circRNA were verified by real time quantitative PCR and the results showed that hsa_circ_0008276, hsa_circ_0003452, hsa_circ_0001846 and hsa_circ_0090508 were down-regulated (P＜0.05), and hsa_circ_0090508 was the more specific than the other three circRNAs. The results of circRNA target genes prediction suggested that has-miR-1294, has-miR-604, has-miR-616, has-miR-663b and has-miR-486-3p may be the potential target genes of hsa_circ_0090508. @*Conclusion@#The differentially expressed circRNA hsa_circ_0090508 was significantly downregulated in the patients with tuberculosis and may affect the regulation mechanism of tuberculosis through target genes.
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Objective: To verify whether EFNB2 gene is targeted by miRNA-497 using molecular biology methods. Methods: Bioinformatic method predicted that EFNB2 gene is targeted by miRNA-497. PCR methods amplified the fragment in EFNB2 gene 3'-UTR including the putative miRNA-497 binding site. Then the sequences of wide type (WT) and mutant (MUT) were cloned into the pmirGLO luciferase vector, respectively. The DNA sequences of the amplified fragments were identified by restriction enzyme digestion and sequencing, and were consistent with the reference sequence from UCSC. This constructed vector was marked as pmirGLO-EFNB2 vector. Finally, the pmirGLO vector, the pmirGLO-EFNB2 vector, miRNA-497 mimics and negative control (NC) were divided into five groups and transfected into HEK393T cells, and the luciferase activity was tested after 24 h and 48 h by dual luciferase reporter gene assay. Results: The results of DNA sequencing demonstrated that the PCR fragment was successfully cloned into pmirGLO vector. The transfection results showed that the recombinant plasmid of WT and MUT was successfully transfected into HEK293T. The results of dual luciferase activity assay demonstrated that miRNA-497 significantly decreased the reporter gene activity compared with the NC. Conclusion: At the cellular level, the schizophrenia susceptibility gene EFNB2 was verified to be targeted by miRNA-497, which provides a new idea and new clue for subsequent studies on miRNA-497 function in the molecular mechanism of schizophrenia.
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Objeetive Airway remodeling is an important pathological feature of asthma.This study is to investigate the role of microRNA-21 (miR-21) in airway remodeling in asthmatic mice.Methods A total of 16 female BALB/c mice were randomly divided into control group and asthma model group.Mice were sensitized and challenged by ovalbumin to establish a murine model of asthma.The mice in the normal control group were intraperitoneally injected with phosphate buffered saline for sensitization and given a phosphate buffered saline inhalation for the challenge.Twenty-four hours after the last aerosol inhalation,lung tissues of mice were sampled and subjectd to Western blot for testing expression of TGFβ type Ⅱ receptor,Smad7,and Collagen Type Ⅰ (COL Ⅰ) in lung tissue of mice of each group.Quantitative real-time PCR was used to test miR-21 expression in lung tissue of mice of each group.Target gene prediction software (TargetScan) was used to predict target gene of miR-21.293T cell was used to conduct dual-luciferase reporter gene assay for verification of miR-21 target gene.Results Compared with control group,asthma group had increased expression of COL Ⅰ protein and miR-21 in lung tissue (t =11.94,P ＜ 0.05;t =23.05,P ＜ 0.05).Smad7 and TGF-βR Ⅱ were target genes of miR-21.Conclusion miR-21 down-regulates Smad7,a target gene of miR-21,activates the TGFβ/Smad signaling pathway,and promotes airway remodeling,indicating that miR-21 may be a therapeutic target for the treatment of airway remodeling in asthma.
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OBJECTIVE To detect the effects of miR-107 on the proliferation, apoptosis and inflammatory factors of human nasal mucosal epithelial cells(HNMECs) induced by Lipopolysaccharides(LPS), and to elucidate the potential molecular mechanism. METHODS The expression changes of miR-107 and HMGBl(High mobility group box 1) in 1 mg/L LPS induced HNMECs before and after LPS induction were detected by using quantitative realtime PCR. LPS induced-HNMECs were transfected with miR-107 agomir and agomir control, and 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide(MTT) assay, flow cytometry and enzyme-linked immunosorbent assay(ELISA) were performed to detect cell proliferation, apoptosis and inflammatory factors. Dual luciferase reporter assay and Western blots were applied to verify whether HMGB1 was a target gene of miR-107. RESULTS The expression of miR-107 in HNMECs after LPS induction was significantly lower than that before induction(t =9.35, P ＜0.05), while the expression of HMGB1 in HNMECs after LPS induction was significantly higher than that before induction(t =13.07, P＜0.05). Compared with transfected agomir control, transfection of miR-107 agomir in HNMECs significantly inhibited cell proliferation (F =17.12, P ＜0.05) and decreased inflammatory factor TNF-α(t =6.11, P ＜0.05). IL-1β(t =6.90, P ＜0.05) and IL-6(t =8.18, P ＜0.05) expression levels, and increased apoptosis rate(t =7.49, P ＜0.05). HMGB1 was a target gene of miR-107, and transfection of miR-107 agomir in HNMECs after LPS induction could significantly reduce the expression of HMGB1 protein (t =28.56, P ＜0.05). CONCLUSION miR-107 is upregulated in HNMECs after LPS induction, while HMGB1 is down-regulated. Overexpression of miR-107 significantly inhibits the proliferation of HNMECs and the expression of inflammatory factors after LPS induction, and promotes apoptosis. HMGB1 is a target gene of miR-107, suggesting that miR-107 may play a regulatory role by regulating HMGB1.