ObjectiveTo explore expression, clinical and biological significance of plasma miRNA-196a from patients with advanced gastric cancer.MethodsReal time quantitative RT-PCR (qRT-PCR) method was used to detect the miRNA-196a levels in tissues and plasma from 75 gastric cancer patients and 35 benign gastric lesions controls. Then clinic pathological correlations of plasma miRNA-196a in 75 gastric cancer patients were analyzed. Twenty-five gastric cancer patients were randomized selected from 75 patients, to compare plasma miRNA-196a levels between preoperation and postoperation. Meanwhile, the effect of miRNA-196a on the invasion ability of gastric cancer MGC-803 cell line was observed in vitro.ResultsThe levels of miRNA-196a in both plasma and tissues from 75 gastric cancer patients were significantly increased compared with 35 benign gastric lesions controls (P<0.000 1). Clinic pathological data of 75 gastric cancer patients showed that the expressions of miRNA-196a were significantly up-regulated in gastric cancer patients with serosal invasion (P<0.001), lymph node metastasis (P=0.004), distant metastasis (P<0.001) and late clinical stage (P<0.001). The expression of miRNA-196a in peripheral plasma of patients with gastric cancer was significantly down regulated after operation (P<0.000 1). In vitro, overexpression of miRNA-196a significantly increased the invasion ability of MGC-803 cells (P<0.05), whereas knockdown of endogenous miRNA-196a significantly inhibited the invasion ability of MGC-803 cells (P<0.05).ConclusionsThe expression of miRNA-196a is up-regulated not only in peripheral plasma of patients with gastric cancer, but also with the progression of gastric cancer (serosal invasion, lymph node metastasis and distant metastasis). The up-regulation of miRNA-196a expression in peripheral plasma is mainly due to the release of primary tumor tissue. miRNA-196a is expected to be a prognostic marker and a potential therapeutic target for advanced gastric cancer.
Objective To screen pyroptosis-related miRNAs of acute aortic dissection (AAD) from the GEO database, and analyze and verify their functions. MethodsThe microarray data set based on the miRNA chip in the GEO database was downloaded, the differentially expressed miRNAs were screened, and the target genes were predicted by the miRWalk database. Pyroptosis-related genes (PRGs) were searched in the PubMed database with "pyroptosis" as the keyword, and the intersection of PRGs and differential miRNAs predicting target genes were taken as AAD PRGs by Venn diagram. GO and KEGG enrichment analyses were performed. CytoHubba was used to screen the critical AAD PRGs and then the AAD pyroptosis-related miRNAs were identified. Aortic tissues were collected from gender- and age-matched AAD patients and healthy people, and the critical PRGs and miRNAs were verified by Western blotting and RT-qPCR. ResultsA total of 46 AAD differentially expressed miRNAs were screened, and 49 AAD PRGs were obtained by Venn diagram. GO enrichment analysis showed that the genes played a vital role in apoptosis regulated by cysteine endopeptidases. KEGG analysis showed that the genes enriched in Salmonella infection, necroptosis, and Nod-like receptor signaling pathways. CytoHubba screened the critical AAD PRGs such as cysteine aspartase-1 (Caspase-1), tumor necrosis factor (IL)-1β, and tumor necrosis factor (TNF), then obtained 12 AAD pyroptosis-related miRNAs. Aortic tissues were collected from 6 AAD patients and 6 healthy people. There were 5 males and 1 females in the AAD group with an average age of 48.70±6.35 years, and 4 males and 2 females in the healty control group with an average age of 45.30±4.58 years. There was no statistical difference between the two groups in terms of gender, age, smoking history, hypertension, diabetes, or coronary heart disease (P>0.05). Western blotting and RT-qPCR results showed that Caspase-1 was up-regulated in the AAD patients' aortic tissues compared with the healthy aorta, and the corresponding miRNAs were miR-198, miR-3202, and miR-514b-5p, which were all down-regulated. Conclusion Through bioinformatics analysis and verification, the critical AAD PRGs are Caspase-1, IL-1β, and TNF, and Caspase-1 is up-regulated and 3 corresponding pyroptosis-related miRNAs are down-regulated, which provides new ideas for the molecular mechanism and targeted therapy of AAD cell pyroptosis.
ObjectiveTo investigate the effect of Smad4 on the fibrosis of tendon derived fibroblasts (TDFs) induced by transforming growth factor β1(TGF-β1) by targeted regulation of miRNA219-5P (miR219-5P). MethodsThe tendons donated by the volunteers were harvested to isolate and culture TDFs. The 3rd generation cells were used for experiment. Chemically synthesized miR219-5P mimics, miR219-5P inhibitor, and negative control sequences were transfected into TDFs. The gene expression of miR219-5P in TDFs was detected by real-time PCR, and the protein expression of Smad4 in TDFs was detected by Western blot at 48 hours after transfection. The combining sites of miR219-5P and Smad4 in 3'UTR district were predicted by informatics software. Wild type and mutant type reporter gene expression vectors were constructed and then targeted verification was carried out by the luciferase reporter gene test. Transfected TDFs were then induced by TGF-β1. The proliferation activity of the cells were measured by the cell counting kit 8 after culturing for 24, 48, and 72 hours. The expressions of fibrosis related proteins in TDFs were detected by Western blot at 72 hours. ResultsAfter TDFs were transfected by miR219-5P mimics, miR219-5P expression was significantly up-regulated, but the expressions of Smad4 was decreased subsequently (P<0.05). Intracellular expression of miR219-5P was inhibited by miR219-5P mimics inhibitor, however, the protein expression of Smad4 was significantly increased (P<0.05). Luciferase reporter gene test showed that luciferase activities were significantly decreased in pGL3-WT-Smad4+mimics group, but were significantly increased in pGL3-WT-Smad4+inhibitor group when compared with pGL3-WT-Smad4 transfected group (P<0.05), but no significant difference was found between GL3-MT-Smad4+mimics and pGL3-MT-Smad4+inhibitor groups (P>0.05). Cell proliferation and the fibrosis related proteins were increased in TGF-β1 induced TDFs, however, decreased in TGF-β1 induced TDFs after transfected by miR219-5P inhibitor (P<0.01). ConclusionmiR219-5P can significantly inhibit fibrosis of TDFs induced by TGF-β1 by down-regulating Smad4 expression.
Colorectal cancer is one of the most common malignant diseases that threatens human being's health. With researches on microRNAs (miRNAs) getting deeper and wider, more evidences revealed that a great many of miRNAs have been involved in the development of colorectal cancer and have the potential to become the biomarker for early diagnosis, prediction of prognosis and recurrence of colorectal cancer. MiRNA-143/miRNA-145 are significantly reduced in several cancers, including colorectal cancer, showing an antitumorigenic activity. In the present article, we make a brief review on the advances in the researches on miRNA-143/miRNA-145 and colorectal cancer to provide guidance for further explorations of the mechanism and target therapy of this disease.
Mechanoresponsive microRNAs are a type of miRNAs which are sensitive or responsive to mechanical strain applied to them, their expression levels were changed after mechanical loading, thus affecting the expression levels of mRNA and proteins they regulated. Up to now, some mechanoresponsive miRNAs have been discovered in physiological or pathological tissues or organs. However, these discoveries are usually limited, and they are not able to guide clinical practice well. According to this situation, this paper summarizes research findings of mechanoresponsive miRNAs, and provides directions for clinical practice and further researches.
Status epilepticus is a life-threatening neurological emergency with high mortality and disability rates. If not treated promptly and effectively, it can lead to severe brain damage or even death. Currently, diagnosis and prognosis assessment mainly rely on electroencephalogram (EEG) and clinical manifestations, which have delays and subjectivity. Therefore, the search for biomarkers that can rapidly, objectively, and accurately assist in the diagnosis, classification, treatment guidance, and prognosis judgment of status epilepticus has become a research hotspot. Biomarkers can reflect the occurrence and development process of the disease at the molecular level, bringing new hope for the precise diagnosis and treatment of status epilepticus. This review aims to systematically elaborate on potential biomarkers in the field of SE.
ObjectiveTo explore the effect of expression of miRNA-21 on bone marrow mesenchymal stem cells (BMSCs).MethodsIn this study, flow cytometry was used to identify the surface-associated antigens of BMSCs. The 10 μmol/L 5-azacytidine was used to induce BMSCs to differentiate to cardiomyocyte-like cells. Immunofluorescence was used to detect the expression of troponin I (cTnI). The samples were assigned to 3 groups: a blank group, a miRNA-21 mimic group, and a negative control (NC) group. The proliferation of BMSCs was detected by methyl thiazolylte-trazolium (MTT), the apoptosis of BMSCs was analyzed by flow cytometry. Western-blotting was used to identify the expression of cTnI and myod in the BMSCs.ResultsThe proliferation of BMSCs was increased, because of the over expression of miRNA-21. But the apoptotic rate of the BMSCs was slower in the miRNA-21 group, on account of the expression of miRNA-21 was higher than that in the NC group and the CK group. The expression of cTnI in the miRNA-21 group was higher than that in the NC group or the CK group.ConclusionThe results suggest that the up-regulation of miRNA-21 enhances proliferation of BMSCs, reduces the apoptosis of BMSCs. miRNA-21 promotes the differentiation of BMSCs, which may pave the way for the treatment directed toward restoring miRNA-21 function for myocardial ischemia.
Objective Hepatitis B virus X (HBx) protein is involved in the initiation and progression of hepatocellular carcinoma (HCC) by regulating the host protein-coding genes. Herein, we want to explore whether HBx protein can alter the expression of microRNAs (miRNAs) to promote proliferation and transformation in malignant hepatocytesin vitro. Methods MiRNA microarray and quantitative reverse-transcription polymerase chain reactions (qRT-PCRs) were performed to identify miRNAs that were differentially regulated by HBx protein in HCC cells. Protein and mRNA expression analyses, cell cycle and apoptosis analyses, and luciferase reporter assays were performed to delineate the consequences of miR-16 family repression in HepG2 cells. Results HBx protein induced widespread deregulation of miRNAs in HepG2 cells, and the downregulation of the miR-16 family was reproducible in HepG2, SK-HEP-1, and Huh7 cells. CCND1, a target gene of the miR-16 family, was derepressed by HBx protein in HepG2 cells. C-myc mediated the HBx-induced repression of miR-15a/16 in HepG2 cells. Ectopically expressed miR-15a/16 suppressed the proliferation, clonogenicity, and anchorage-independent growth of HBx-expressing HepG2 cells by arresting them in the G1 phase and inducing apoptosis, whereas reduced expression of miR-16 accelerated the growth and cell-cycle progression of HepG2 cells. Conclusions HBx protein altered thein vitro expression of miRNAs in host malignant hepatocytes, particularly downregulating the miR-16 family. Repression of miR-15a/16 is c-myc mediated and is required for the HBx-induced transformation of HepG2 cellsin vitro. Therefore, miR-16 family may serve as a therapeutic target for hepatitis B virus (HBV)-associated HCC.
ObjectiveTo explore the dynamic expression changes of neuronal growth and differentiation-associated miR-124a and miR-9 in the process of epileptogenesis. MethodsEstablish the lithium-pilocarpine induced status epilepticus (SE) rat model. Animal behavior change induced by SE as well as in the period of chronic epilepsy was observed by naked-eye or video-recording. Major time points for the study were chosen at 1d, 7d, 14d and 28d post-SE, on which the post-SE rats were decapitated and their hippocampal specimens were obtained. Total RNA from each specimen was extracted and qPCR was exploited to detect miR-124a and miR-9 expression in the specimens. Statistical analysis was used to show the dynamic expressional changes of miR-124a and miR-9 in rat hippocampus at 1d, 7d, 14d and 28d post-SE during the process of epileptogenesis. ResultsCompared with normal rats, the expression level of miR-124a in rat hippocampus did not show a significant difference at 1d post-SE, but it had shown markedly differences at 7d, 14d and 28d post-SE(P < 0.05), with a declining trend. Compared with normal rats, the expression level of miR-9 had demonstrated significant differences at 1d, 7d, 14d and 28d post-SE(P < 0.05)with a generally increasing trend, although there was slight fluctuation of expressional up-regulation at 7d post-SE. ConclusionNeuronal growth and differentiation-associated miR-124a and miR-9 had shown dynamic changes of down-regulation or up-regulation in the process of epileptogenesis. It can be suspected that miR-124a and miR-9 take part in hippocampal neurogenesis post-SE and be involved in epileptogenesis process.
ObjectiveTo systematically review the diagnostic value of miRNAs for Alzheimer’s disease (AD).MethodsPubMed, Web of Science, EMbase, The Cochrane Library, CNKI, WanFang Data, VIP, and CBM databases were electronically searched to collect diagnostic tests of miRNAs for AD from inception to October 31, 2020. Two researchers independently screened literature, extracted data, and assessed the risk of bias of the included studies. RevMan 5.3 and Stata 14.0 software were used for meta-analysis. ResultsA total of 22 studies involving 4 006 subjects were included. The meta-analysis results showed that the pooled sensitivity, specificity, positive and negative likelihood ratios, diagnostic odds ratio, and the areas under the working characteristic curve of miRNA in AD diagnosis were 0.83 (95%CI 0.79 to 0.87), 0.80 (95%CI 0.76 to 0.83), 4.07 (95%CI 3.37 to 4.92), 0.21 (95%CI 0.17 to 0.27), 19.20 (95%CI 12.96 to 28.48) and 0.88 (95%CI 0.85 to 0.90), respectively. ConclusionThe current evidence shows that miRNAs have a high diagnostic value for AD. However, because of the limited quality and quantity of the included studies, more high-quality studies are required to verify the above conclusion.