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1.
Braz. j. med. biol. res ; 54(2): e9869, 2021. tab, graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1142585

ABSTRACT

Severe blockage in myeloid differentiation is the hallmark of acute myeloid leukemia (AML). Trdmt1 plays an important role in hematopoiesis. However, little is known about the function of Trdmt1 in AML cell differentiation. In the present study, Trdmt1 was up-regulated and miR-181a was down-regulated significantly during human leukemia HL-60 cell differentiation after TAT-CT3 fusion protein treatment. Accordingly, miR-181a overexpression in HL-60 cells inhibited granulocytic maturation. In addition, our "rescue" assay demonstrated that Trdmt1 3′-untranslated region promoted myeloid differentiation of HL-60 cells by sequestering miR-181a and up-regulating C/EBPα (a critical factor for normal myelopoiesis) via its competing endogenous RNA (ceRNA) activity on miR-181a. These findings revealed an unrecognized role of Trdmt1 as a potential ceRNA for therapeutic targets in AML.


Subject(s)
Humans , Leukemia, Myeloid, Acute/genetics , MicroRNAs/genetics , DNA (Cytosine-5-)-Methyltransferases/genetics , Cell Differentiation , HL-60 Cells
2.
Article in English | WPRIM | ID: wpr-880866

ABSTRACT

As an important component of the tumor microenvironment, cancer-associated fibroblasts (CAFs) secrete energy metabolites to supply energy for tumor progression. Abnormal regulation of long noncoding RNAs (lncRNAs) is thought to contribute to glucose metabolism, but the role of lncRNAs in glycolysis in oral CAFs has not been systematically examined. In the present study, by using RNA sequencing and bioinformatics analysis, we analyzed the lncRNA/mRNA profiles of normal fibroblasts (NFs) derived from normal tissues and CAFs derived from patients with oral squamous cell carcinoma (OSCC). LncRNA H19 was identified as a key lncRNA in oral CAFs and was synchronously upregulated in both oral cancer cell lines and CAFs. Using small interfering RNA (siRNA) strategies, we determined that lncRNA H19 knockdown affected proliferation, migration, and glycolysis in oral CAFs. We found that knockdown of lncRNA H19 by siRNA suppressed the MAPK signaling pathway, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) and miR-675-5p. Furthermore, the lncRNA H19/miR-675-5p/PFKFB3 axis was involved in promoting the glycolysis pathway in oral CAFs, as demonstrated by a luciferase reporter system assay and treatment with a miRNA-specific inhibitor. Our study presents a new way to understand glucose metabolism in oral CAFs, theoretically providing a novel biomarker for OSCC molecular diagnosis and a new target for antitumor therapy.


Subject(s)
Cancer-Associated Fibroblasts/metabolism , Carcinoma, Squamous Cell/genetics , Cell Line, Tumor , Cell Proliferation , Gene Expression Regulation, Neoplastic , Glycolysis , Head and Neck Neoplasms , Humans , MicroRNAs/metabolism , Mouth Neoplasms/genetics , Phosphofructokinase-2/genetics , RNA, Long Noncoding/genetics , Signal Transduction , Tumor Microenvironment
3.
Article in Chinese | WPRIM | ID: wpr-880841

ABSTRACT

OBJECTIVE@#To investigate the effects of overexpression of long noncoding RNA (lncRNA) MEG3 on the proliferation and invasion of glioblastoma U251 cells by suppressing the expression of hypoxia inducible factor 1@*METHODS@#The expression of lncRNA MEG3 and HIF1@*RESULTS@#The expression of MEG3 was significantly lower and HIF1@*CONCLUSIONS@#MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1


Subject(s)
Apoptosis , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , Glioblastoma/genetics , Humans , MicroRNAs , RNA, Long Noncoding/genetics
4.
Article in English | WPRIM | ID: wpr-880659

ABSTRACT

Epstein-Barr virus (EBV), a definite tumorigenic virus, is closely related to the development of nasopharyngeal cancer, gastric cancer, lymphoma and other tumors. EBV encodes a total of 44 mature microRNAs, which can regulate the expression of virus and host genes. EBV-encoded microRNAs and their regulated target molecules participate in the biological functions of tumor apoptosis, proliferation, invasion, and metastasis during tumorigenesis and development, and play an important role in the development of tumor.


Subject(s)
Carcinogenesis/genetics , Epstein-Barr Virus Infections/genetics , Gene Expression Regulation, Neoplastic , Herpesvirus 4, Human/genetics , Humans , MicroRNAs/genetics , Nasopharyngeal Neoplasms/genetics
5.
Article in English | WPRIM | ID: wpr-880617

ABSTRACT

OBJECTIVES@#To investigate the effects of propofol on the proliferation and invasion of glioma U87 cells and to explore the possible anti-tumor mechanisms.@*METHODS@#The glioma U87 cells was divided into a blank group, a positive control group, and the propofol groups (1.00, 2.00 or 5.00 mmol/L). Cell counting kit-8 (CCK-8) was used to detect cell proliferation; Transwell method was used to detect the effect of propofol on invasion and migration of U87 cells; real-time PCR was used to detect the expression of microRNA-134 (miR-134); Western blotting was used to detect the expression levels of reproduction-related protein Ki-67, invasion-related protein metalloproteinase-2 (MMP-2), metalloproteinase-9 (MMP-9) and phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) signaling pathway-related protein.@*RESULTS@#Compared with the blank group, the proliferation, invasion and migration capacity of U87 cells were reduced in the positive control group and the propofol groups after 48 hours (all @*CONCLUSIONS@#Propofol can decrease the proliferation rate, and the invasion and migration abilities of U87 cells, which may be achieved by up-regulation of miR-134 and suppression of PI3K/Akt signaling pathway.


Subject(s)
Cell Line, Tumor , Cell Movement , Cell Proliferation , Glioma/genetics , Humans , Matrix Metalloproteinase 2/genetics , MicroRNAs/genetics , Phosphatidylinositol 3-Kinases/genetics , Propofol/pharmacology , Proto-Oncogene Proteins c-akt/genetics
6.
Article in English | WPRIM | ID: wpr-880615

ABSTRACT

OBJECTIVES@#To study the gene expression of adipose tissue CD14@*METHODS@#The data of GSE54350 were obtained from the public database of gene expression profiling. The data were pre-processed by Network Analyst, String 11.0, Cytoscape 3.7.1, and other analytical software. The differentially expressed genes were analyzed by gene ontology biological function and kyoto encycopedia of genes and genomes (KEGG) signaling pathway to establish differential gene protein interaction network, transcription factor-gene regulatory network, microRNA-gene regulatory network, environmental factors-gene regulatory network, and other interaction systems.@*RESULTS@#The gene expression pattern of CD14@*CONCLUSIONS@#The gene expression of adipose tissue CD14


Subject(s)
Adipose Tissue , Computational Biology , DNA-Binding Proteins , Diabetes Mellitus, Type 2/genetics , Gene Expression , Gene Expression Profiling , Gene Regulatory Networks , Humans , MicroRNAs/genetics , Muscle Proteins
7.
Article in Chinese | WPRIM | ID: wpr-880175

ABSTRACT

OBJECTIVE@#To investigate the clinical correlation of expression level changes of miR-181b and miR-194 to the pathogenesis of acute graft-versus-host disease (aGVHD), and determine plasma miR-181b and miR-194 as the potential biomarkers for aGVHD.@*METHODS@#The plasma samples were collected from 31 patients who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) at before HSCT, 15 days after HSCT and onset of aGVHD. The expression levels of miR-181b and miR-194 were detected by quantitative real-time PCR. Receiver-operating characteristic (ROC) curves and the area under the ROC curve (AUC) were used to assess the sensitivity and specificity of miRNA biomarkers for the diagnosis of aGVHD.@*RESULTS@#MiR-181b and miR-194 downregulated after treatment were significantly upregulated in the plasma at onset of aGVHD (P0.05). The expressions of plasma miR-181b and miR-194 collected on day 15 after HSCT were significantly upregulated in the patients with aGVHD in comparison with non-GVHD patients (P<0.05). Moreover, these elevated miRNAs were detected before aGVHD. The AUC of miR-181b predicting aGVHD was 0.91±0.05 (specificity was 0.94, sensitivity was 0.69). The AUC of miR-194 predicting aGVHD was 0.91±0.06 (specificity was 0.94, sensitivity was 0.77).@*CONCLUSION@#MiR-181b and miR-194 may serve as early biomarkers for the diagnosis and prognosis of aGVHD.


Subject(s)
Acute Disease , Biomarkers , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Humans , MicroRNAs , Transplantation, Homologous
8.
Article in Chinese | WPRIM | ID: wpr-880144

ABSTRACT

OBJECTIVE@#To investigate the relationship between the polymorphism of miR-155 and its target gene MyD88 and clinicopathological features of diffuse large B-cell lymphoma (DLBCL).@*METHODS@#135 cases of DLBCL patients in our hospital from March 2015 to August 2017 were selected, and 90 cases of reactive hyperplasia of lymph nodes were selected as the control group. The relative expression of miR-155 and MyD88 gene polymorphism were detected in the two groups, and the relationship between miR-155 and MyD88 gene polymorphism and clinicopathological characteristics of DLBCL was analyzed.@*RESULTS@#The relative expression of miR-155 in DLBCL patients was significantly higher than that in the control group (P<0.05). The mutation rate of MyD88 L265P in DLBCL group was significantly higher than that in control group (P<0.05). The relative expression of miR-155 in patients with MyD88 L265P mutation was significantly higher than that in patients with wild-type DLBCL (P<0.05). The relative expression of miR-155 and the polymorphism of MyD88 L265P were associated with lesion location, stage, BCL-2 protein expression and MyD88 protein expression in DLBCL patients (t=7.461、8.804、6.487、10.812; χ@*CONCLUSION@#The abnormal expression of miR-155 and the mutation rate of MyD88 gene in DLBCL patients are increased, and the expression of miR-155 and the mutation of MyD88 gene affect the disease progression and prognosis of patients, which may be potential biological indicators for the diagnosis, treatment and prognosis of DLBCL.


Subject(s)
Humans , Lymphoma, Large B-Cell, Diffuse/genetics , MicroRNAs/genetics , Mutation , Myeloid Differentiation Factor 88/genetics , Polymorphism, Genetic , Prognosis
9.
Article in Chinese | WPRIM | ID: wpr-880095

ABSTRACT

OBJECTIVE@#To investigate the expression of microRNA-370 (miR-370) and microRNA-203 (miR-203) in the serum of patients with acute myeloid leukemia(AML), and to analyze its clinical diagnosis and prognostic significance.@*METHODS@#57 patients with acute myeloid leukemia were enrolled as experimental group, and 21 healthy people were enrolled as control group. The fasting venous blood of the personal in the two groups were collected. The expression of miR-370 and miR-203 of the personal in each groups were detected by real-time fluorescent quantitative PCR. The receiver operating characteristic (ROC) curve was plotted to detected the diagnostic values of serum miR-370, miR-203, and the Kaplan-Meier method was used to estimate the relationship between expression and overall survival of the patients.@*RESULTS@#Compared with healthy controls, serum miR-370 expression was significantly decreased in AML patients(P<0.05), and serum miR-203 expression was also significantly decreased (P<0.05). ROC curve analysis showed that the expression of serum miR-370 and miR-203 could be used to distinguish acute myeloid leukemia and healthy people. The area under the ROC curve of miR-370 was 0.909, and the sensitivity and specificity were 91.46% and 100.00%, respectively. The area under the ROC curve of miR-203 was 0.895, and the sensitivity and specificity were 83.45% and 89.71%, respectively. Serum levels of miR-370 and miR-203 were closely related to overall survival in AML patients.@*CONCLUSION@#The expression of miR-370 and miR-203 is decreased in the serum of patients with AML and may be a new markers for the diagnosis and prognosis of AML.


Subject(s)
Biomarkers, Tumor , Humans , Kaplan-Meier Estimate , Leukemia, Myeloid, Acute/genetics , MicroRNAs , Prognosis , ROC Curve
10.
Article in Chinese | WPRIM | ID: wpr-880094

ABSTRACT

OBJECTIVE@#To analyze the relationship of the expression of transcription factor MYB targeted regulation by miR-96 to cell invasion and apoptosis in pediatric acute myeloid leukemia (AML).@*METHODS@#A total of 65 children with AML in The 928 Hospital of PLA Joint Logistics Support Forces from January 2017 to November 2019 were selected, including 35 cases diagnosed as primary AML and 30 cases as complete remission AML. Thirty children with immune thrombocytopenia were selected as control group. The clinical characteristics were analyzed and compared between the two groups. The levels of miR-96 and MYB in peripheral blood samples were detected by qRT-PCR and compared between the two groups. The miR-96 mimics and its negative control (NC), inhibitor-miR-96 and its NC transfected HL60 cells induced by liposome (Lipofectamine 2000), respectively, Then the expression levels of MYB were detected with Western blot and compared among four HL60 cell groups. The invasion ability of four HL60 cell groups were detected with Transwell assay. The cell proliferation ability of four HL60 cell groups were detected with MTT at 24 h, 48 h, and 72 h, respectively. The apoptosis rates of four HL60 cell groups were detected with flow cytometry.@*RESULTS@#Compared with control group, the level of miR-96 in AML children were higher, but MYB lower (P0.05). The promotion of over-expression level of miR-96 on the invasion ability of HL 60 cells was confirmed by Transwell assay. MTT assay showed that miR-96 could promote the proliferation of HL60 cells, inhibit the apoptosis of HL60 cells, and the effect was time-dependent manner (r=0.804). The inhibition of miR-96 on HL60 cells apoptosis was also confirmed with flow cytometry.@*CONCLUSION@#MiR-96 has significant negative effect on invasion and apoptosis of AML cells by targeting regulation MYB, and it might be a potential novel strategy for pediatric AML treatment.


Subject(s)
Apoptosis , Cell Line, Tumor , Cell Proliferation , Child , HL-60 Cells , Humans , Leukemia, Myeloid, Acute/genetics , MicroRNAs/genetics , Proto-Oncogene Proteins c-myb
11.
Article in Chinese | WPRIM | ID: wpr-880079

ABSTRACT

OBJECTIVE@#To identify differentiation related miRNA and evaluate roles of miRNA during ATRA induced myeloid differentiation.@*METHODS@#The small RNA sequencing was used to analyze differential expressed miRNAs in ATRA induced NB4 cells. Then the several up or down-regulated miRNA were selected as the research candidates. SgRNAs targeting the genome of each miRNA were designed and NB4 cells with inducible expression of Cas9 protein were generated. After transduced sgRNA into NB4/Cas9 cells, the mutation level by PCR and surveyor assay were evaluated. The cell differentiation level was investigated by surface CD11b expression via flow cytometry.@*RESULTS@#A total of 410 mature miRNAs which expressed in NB4 cells were detected out after treated by ATRA, 74 miRNAs were up-regulated and 55 were down-regulated miRNAs with DNA cleavage generated by CRISPR/Cas9 was assayed directly by PCR or surveyor assay, quantitative PCR showed that the expression of miRNA was downregulated, which evaluated that gene edition successfully inhibitied the expression of mature miRNA. MiR-223 knockout showed the myeloid differentation of NB4 significantly inhibitied, while miRNA-155 knockout showed the myeloid differentation of NB4 cells significantly increased.@*CONCLUSION@#CRISPR/Cas9 is a powerful tool for gene editing and can lead to miRNA knockout. Knockouts of miR-223 and miR-155 have shown a differentiation-related phenotype, and the potential mechanism is the integrative regulation of target genes.


Subject(s)
CRISPR-Cas Systems , Cell Differentiation , Gene Editing , MicroRNAs/genetics , Sequence Analysis, RNA , Tretinoin
12.
Article in Chinese | WPRIM | ID: wpr-879620

ABSTRACT

OBJECTIVE@#To construct the differential expression profile of microRNA (miRNA) in plasma of patients with type 2 diabetes mellitus (T2DM) and explore the possibility of using miRNA as the target for diagnosis and treatment of T2DM.@*METHODS@#Agilent miRNA microarray was used to determine the expression profiles of miRNA in the plasma of patients with T2DM (FC> 2, P< 0.05). The result was verified by real-time quantitative PCR (RT-qPCR). Candidate miRNA was analyzed by bioinformatic tools.@*RESULTS@#In total 122 differentially expressed miRNAs were identified. Among these, 14 were selected by multi-source intersection screening, which included 5 up-regulated genes and 9 down regulated genes. RT-qPCR showed that the expression of hsa-miR-185-5p and hsa-miR-328-5p have significantly increased in T2DM patients (P< 0.05). Bioinformatic analysis suggested that these miRNAs may be involved in the pathogenesis of T2DM through insulin secretion and PI3K-AKT signaling pathway.@*CONCLUSION@#Differential expression of hsa-miR-185-5p and hsa-miR-328-5p in the plasma may be closely associated with the pathogenesis of T2DM.


Subject(s)
Computational Biology , Diabetes Mellitus, Type 2/genetics , Humans , MicroRNAs/genetics , Phosphatidylinositol 3-Kinases , Signal Transduction
13.
Chinese Journal of Biotechnology ; (12): 663-672, 2021.
Article in Chinese | WPRIM | ID: wpr-878591

ABSTRACT

We developed a high-efficiency microfluidic chip for extracting exosomes from human plasma. We collected peripheral blood from normal human, designed and fabricated a microfluidic chip based on nanoporous membrane and agarose gel electrophoresis to isolate exosomes. The extracted exosomes were characterized by transmission electron microscopy, nanosight and Western blotting, the morphology, concentration and particle size of exosomes were identified and analyzed. Meanwhile, we used ultracentrifugation and microfluidic chip to isolate exosomes separately. The particle size and concentration of the exosomes extracted by two methods were compared and analyzed, and their respective extraction efficiency was discussed. Finally, the expression level of miRNA-21 in exosomes was analyzed by RT-PCR. The microfluidic chip isolated (in 1 hour) high-purity exosomes with size ranging from 30-200 nm directly from human plasma, allowing downstream exosomal miRNA analysis. By comparing with ultracentrifugation, the isolation yield of microfluidic chip was 3.80 times higher than ultracentrifugation when the volume of plasma sample less than 100 μL. The optimized parameters for exosome isolation by gel electrophoresis microfluidic chip were: voltage: 100 V; concentration of agarose gel: 1.0%; flow rate of injection pump: 0.1 mL/h. The gel electrophoresis microfluidic chips could rapidly and efficiently isolate the exosomes, showing great potential in the research of exosomes and cancer biomarkers.


Subject(s)
Exosomes , Humans , MicroRNAs/genetics , Microfluidics , Plasma , Ultracentrifugation
14.
Article in Chinese | WPRIM | ID: wpr-878422

ABSTRACT

OBJECTIVES@#This study aims to construct endogenous exosomes abundantly loaded with miR-1 and investigate the role of exosome-mediated microRNA-1 (miR-1) delivery on CAL-27 cell proliferation.@*METHODS@#Exosomes secreted by miR-1-overexpressing HEK293 cells (miR1-EXO) were purified via ultracentrifugation and subjected to transmission electron microscopy, nanoparticle analysis, Western blot analysis, and quantitative polymerase chain reaction (qPCR). CAL-27 cells were cocultured with exosomes secreted by HEK293 cells (CON-EXO) and miR1-EXO and equivalent phosphate buffer saline. The intracellular transport of exosomes was measured by using immunofluorescence, the expression of miR-1 and its target gene MET were investigated via qPCR, CAL-27 cell proliferation was measured through MTT assay, and cell cycle state was determined by applying flow cytometry.@*RESULTS@#Electron microscopy revealed that miR1-EXO and CON-EXO were spherical or cup-shaped with an average diameter of approximately 110 nm. The well-known exosome markers CD9, Tsg101, and Alix were enriched. The expression of miR-1 in miR1-EXO was higher than that in CON-EXO (285.80±14.33 vs 1.00±0.06, @*CONCLUSIONS@#Exosomes secreted from miR1-EXO cells could load abundant miR-1. Exosomal miR-1 delivered into CAL-27 cells by using miR1-EXO suppressed the expression of MET mRNA and inhibited cell proliferation.


Subject(s)
Cell Cycle , Cell Proliferation , Exosomes , HEK293 Cells , Humans , MicroRNAs
15.
Article in English | WPRIM | ID: wpr-878405

ABSTRACT

OBJECTIVES@#This study aimed to investigate the effects of microRNA-146a (miR-146a) on the production of cytokines in lymphocytes stimulated by @*METHODS@#Lymphocytes were harvested from mouse spleen and cultured @*RESULTS@#Compared with non-LPS-stimulated group, @*CONCLUSIONS@#MiR-146a can provide a suitable microenvironment for bone formation by preventing the inflammatory effects of


Subject(s)
Animals , Cytokines , Lipopolysaccharides , Lymphocytes , Mice , MicroRNAs , Porphyromonas gingivalis
16.
Article in English | WPRIM | ID: wpr-878404

ABSTRACT

OBJECTIVES@#To study the hypoxia response gene and microRNA (miRNA) expression profiles in the pathogenesis and progression of oral leukoplakia (OLK).@*METHODS@#Affymetrix GeneChip human transcriptome array 2.0 was used to detect the transcriptome of normal mucosa, low-risk OLK, high-risk OLK, and early squamous cell carcinoma (SCC). Gene ontology function analysis was used to screen genes and key miRNAs whose biological role is hypoxia response. Quantitative reverse transcription polymerase ch-ain reaction (qRT-PCR) was used to verify the expression of hypoxia response genes and miRNAs.@*RESULTS@#A total of 7 different genes of hypoxia response between normal mucosa and low-risk OLK, 10 genes between low-risk and high-risk OLK, and 21 genes between high-risk OLK and SCC were identified. The results of qRT-PCR showed that the expression of hypoxia-inducible factor 1α, chemokine cc-motif ligand 2, and matrix metalloproteinase 3 mRNA and miR-21 in normal mucosa, OLK, and SCC increased in a stepwise manner. The expression difference between OLK and SCC was statistically significant and consistent with the results of transcriptome array.@*CONCLUSIONS@#The hypoxia response gene and related miRNA play roles in the development and progression of OLK.


Subject(s)
Carcinogenesis , Humans , Hypoxia , Leukoplakia, Oral , MicroRNAs , Mouth Neoplasms , Transcriptome
17.
Article in English | WPRIM | ID: wpr-878339

ABSTRACT

Objective@#Cervical cancer (CC) is one of the most common malignant tumors in gynecology. This study aimed to investigate the prognostic significance of serum microRNA (miR)-378a-3p in CC and the effect of miR-378a-3p on tumor growth.@*Methods@#Real-time quantitative polymerase chain reaction analysis was used to measure the expression of miR-378a-3p in serum from patients with CC and healthy control subjects as well as from CC tissues and adjacent normal tissues. The association between serum miR-378a-3p levels and clinicopathological factors was analyzed. The correlation between miR-378a-3p levels and overall survival (OS) of CC patients was determined by Kaplan-Meier analysis. The CC cell proliferation and migration abilities after transfection of miR-378a-3p mimics were detected by Cell Counting Kit-8 and scratch wound healing assays, respectively. Tumor volume and weight in mice treated with miR-378a-3p were measured using a caliper and an electronic balance.@*Results@#MiR-378a-3p expression was downregulated in the serum and tissues of CC patients compared to that in healthy control subjects and normal tissues, respectively. Low expression of miR-378a-3p was positively correlated with large tumor size, advanced tumor stage, and lymph node metastasis. The OS of patients with low expression of miR-378a-3p was significantly lower than that of patients with high expression. Overexpression of miR-378a-3p suppressed the proliferation and migration of CC cells. @*Conclusion@#MiR-378a-3p downregulation is associated with the development and prognosis of CC, suggesting that it may be a potential biomarker for CC.


Subject(s)
Animals , Biomarkers/blood , Cell Movement , Cell Proliferation , Down-Regulation , Female , Gene Expression Regulation, Neoplastic , Humans , Mice , Mice, Inbred BALB C , MicroRNAs/blood , Middle Aged , Uterine Cervical Neoplasms/metabolism
18.
Article in English | WPRIM | ID: wpr-878317

ABSTRACT

Objective@#In the present study, the ABCA1 was used as a label to capture specific exosomes, the level of ABCA1-labeled exosomal microRNA-135a (miR-135a) was evaluated for the diagnosis of Alzheimer's disease (AD), especially in patients with early stages of AD.@*Methods@#This is a preliminary research focused on the levels of ABCA1 in WBCs, RBCs, HT-22 cells, and neuron cells. The diagnostic value of ABCA1-labeled exosomal miR-135a was examined using the CSF and serum of APP/PS1 double transgenic mice, and 152 patients with SCD, 131 patients with MCI, 198 patients with DAT, and 30 control subjects.@*Results@#The level of ABCA1 exosomes harvested from HT-22 cells and neuron culture medium was significantly higher compared to that of RBCs and WBCs ( @*Conclusion@#This study outlines a method to capture specific exosomes and detect them using immunological methods, which is more efficient for early diagnosis of AD.


Subject(s)
ATP Binding Cassette Transporter 1/cerebrospinal fluid , Aged , Aged, 80 and over , Alzheimer Disease/cerebrospinal fluid , Animals , Biomarkers/cerebrospinal fluid , Cell Line , Cognitive Dysfunction/cerebrospinal fluid , Erythrocytes/metabolism , Exosomes , Female , Humans , Leukocytes/metabolism , Male , Mice, Transgenic , MicroRNAs/blood , Neurons/metabolism
19.
Acta Physiologica Sinica ; (6): 253-262, 2021.
Article in Chinese | WPRIM | ID: wpr-878254

ABSTRACT

The aim of this study was to investigate the effects of dexmedetomidine (Dex) on hepatic ischemia/reperfusion injury (HIRI) and the underlying mechanism. The in vitro HIRI was induced by culturing HL-7702 cells, a human hepatocyte cell line, under 24 h of hypoxia and 12 h of reoxygenation. Quantitative real time PCR (qRT-PCR) and Western blot were performed to detect the expression levels of long non-coding RNA MALAT1, microRNA-126-5p (miR-126-5p) and high mobility group box-1 (HMGB1). Bioinformatics prediction and double luciferase assay were used to verify the targeting relationship between miR-126-5p and MALAT1, HMGB1. Reactive oxygen species (ROS), malondialdehyde (MDA) and ATP levels in culture medium were detected by corresponding kits. The results showed that Dex significantly reduced the levels of ROS and MDA, but increased the level of ATP in HL-7702 cells with HIRI. HIRI up-regulated the expression levels of MALAT1 and HMGB1, and down-regulated the level of miR-126-5p. Dex reversed these effects of HIRI. Furthermore, Dex inhibited HIRI-induced cellular apoptosis, whereas MALAT1 reversed the effect of Dex. This inhibitory effect of Dex could be restored by up-regulation of miR-126-5p. The results suggest that Dex protects hepatocytes from HIRI via regulating MALAT1/miR-126-5p/HMGB1 axis.


Subject(s)
Dexmedetomidine/pharmacology , HMGB1 Protein/genetics , Humans , MicroRNAs/genetics , RNA, Long Noncoding/genetics , Reperfusion Injury/genetics
20.
Chinese Medical Journal ; (24): 564-572, 2021.
Article in English | WPRIM | ID: wpr-878081

ABSTRACT

BACKGROUND@#The pathogenesis of osteosarcoma (OS) is still unclear, and it is still necessary to find new targets and drugs for anti-OS. This study aimed to investigate the role and mechanism of the anti-OS effects of miR-296-5p.@*METHODS@#We measured the expression of miR-296-5p in human OS cell lines and tissues. The effect of miR-296-5p and its target gene staphylococcal nuclease and tudor domain containing 1 on proliferation, migration, and invasion of human OS lines was examined. The Student's t test was used for statistical analysis.@*RESULTS@#We found that microRNA (miR)-296-5p was significantly downregulated in OS cell lines and tissues (control vs. OS, 1.802 ± 0.313 vs. 0.618 ± 0.235, t = 6.402, P < 0.01). Overexpression of miR-296-5p suppressed proliferation, migration, and invasion of OA cells. SND1 was identified as a target of miR-296-5p by bioinformatic analysis and dual-luciferase reporter assay. Overexpression of SND1 abrogated the effects induced by miR-296-5p upregulation (miRNA-296-5p vs. miRNA-296-5p + SND1, 0.294 ± 0.159 vs. 2.300 ± 0.277, t = 12.68, P = 0.003).@*CONCLUSION@#Our study indicates that miR-296-5p may function as a tumor suppressor by targeting SND1 in OS.


Subject(s)
Bone Neoplasms/genetics , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Endonucleases/genetics , Gene Expression Regulation, Neoplastic , Genes, Tumor Suppressor , Humans , MicroRNAs/genetics , Osteosarcoma/genetics
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