Inhibition Of miR-128 Research Articles

LncRNA FOXD3-AS1 aggravates myocardial ischemia/reperfusion injury by inactivating the Redd1/AKT/GSK3β/Nrf2 signaling pathway via the miR-128/TXNIP axis.

Long noncoding RNA forkhead box D3-antisense RNA 1(FOXD3-AS1) is associated with cardiovascular diseases, but its roles in myocardial ischemia/reperfusion (I/R) injury and the related signaling pathway have not been fully reported. We aimed to investigate the roles and mechanism of action of FOXD3-AS1 in myocardial I/R injury. An in vivo myocardial I/R injury mouse model and an in vitro hypoxia/reoxygenation (H/R) cardiomyocyte model was established. Quantitative reverse transcription-polymerase chain reaction, western blotting, and immunofluorescent assays were performed to examine the expression levels of FOXD3-AS1,microRNA (miR)-128,thioredoxin-interacting protein/regulation of development and DNA damage response 1/protein kinase B/glycogen synthase kinase 3β/nuclear factor erythroid 2-related factor 2(TXNIP/Redd1/AKT/GSK3β/Nrf2) pathway-related proteins and apoptosis-related proteins. The interactions between FOXD3-AS1 and miR-128 and miR-128 and TXNIP were analyzed by Spearman's correlation test, predicted by ENCORI, and verified by dual-luciferase reporter assay. In addition, the levels of cardiac injury markers and oxidative stress markers were evaluated by corresponding kits. Cell Counting Kit-8 assays and flow cytometry were performed to assess cell viability and apoptosis. Hematoxylin and eosin staining was applied to observe the effect of FOXD3-AS1 on the morphology of myocardial I/R injured tissues. The results showed that the FOXD3-AS1 and TXNIP were highly expressed, whereas miR-128 was expressed at low levels in I/R myocardial tissues and H/R-induced H9c2 cells. FOXD3-AS1 directly targeted miR-128 to reduce its expression. TXNIP was confirmed as a downstream target of miR-128. Knockdown of FOXD3-AS1 led to the alleviation of I/R injury in vivo and in vitro. FOXD3-AS1enhanced the expression of TXNIP by sponging miR-128, which inhibited the Redd1/AKT/GSK3β/Nrf2 pathway. Both inhibition of miR-128 and overexpression of TXNIP reversed the cardioprotective effect of FOXD3-AS1 small interferingRNA in H/R-induced H9c2 cells.

RETRACTED ARTICLE: microRNA-128 mediates CB1 expression and regulates NF-KB/p-JNK axis to influence the occurrence of diabetic bladder disease

BackgroundDiabetic bladder disease is common complications of diabetes, its symptoms are diverse, can be due to different stages. In this study we investigate the mechanism of miR-128 targeting CB1 expression to mediate the occurrence of diabetic bladder disease.MethodsBioinformatics analysis predicts related regulatory factors of miR-128 in diabetic bladder disease. Models of diabetic bladder lesions were constructed in male SD rats by intraperitoneal injection of streptozotocin at 65 mg/kg body weight. The expression of miR-128 and CB1 mRNA in bladder tissues of each group was detected by RT-qPCR, and CB1, NF-KB, p-JNK and Bcl2 protein expression was detected by Western Blotting. We tested the function of the bladder by urodynamics, detected the pathological characteristics of the bladder tissue by HE staining, and verified the targeting relationship between miR-128 and CB1 through the prediction of the biological website, dual luciferase reporter gene assay and RIP.ResultsmiR-128 was highly expressed in the bladder tissue of diabetic rats. Inhibition of miR-128 could improve the occurrence of diabetic bladder lesions in rats. miR-128 could target the inhibition of CB1 expression, and high expression of CB1 could antagonize miR-128 against diabetic bladder. In the diabetic bladder, miR-128 can regulate the expression of NF-KB and p-JNK through CB1 and affect the level of apoptosis. miR-128 regulates NF-KB/p-JNK through CB1, thus affecting the occurrence of diabetic bladder disease.ConclusionThe high expression of miR-128 can down-regulate the expression of CB1, promote the activation of NF-KB and p-JNK, increase the level of apoptosis and promote the occurrence of diabetic bladder disease.

MicroRNA-128 is involved in dexamethasone-induced lipid accumulation via repressing SIRT1 expression in cultured pig preadipocytes

In this study, pig preadipocytes were firstly treated with 10−6 M DEX for 48 h to explore the role of dexamethasone (DEX, a chemically synthesized long-acting glucocorticoid) on lipid accumulation. Then, miRNA scrambled control (miR-SC), miR-128 overexpression plasmid and miR-128 inhibitor were respectively transfected into pig preadipocytes at 24 h before DEX treatment for 48 h (miR-SC-DEX, miR-128-DEX and miR-128-inhibitor-DEX) to illustrate the regulatory role of miR-128 on DEX-induced lipid accumulation. Compared with control preadipocytes, 10-6 M Dex significantly increased triglyceride (TG) level, whereas the cell proliferation did not change. Moreover, 10-6 M Dex obviously decreased sirtuin 1 (SIRT1) and its related lipolysis genes adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL) mRNA expression and enzyme activity, while significantly increased expression of adipogenesis genes peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α (C/EBP-α) and fatty acid synthase (FAS). In addition, 10-6 M DEX significantly upregulated miR-128 expression, which was confirmed to directly target SIRT1 by bioinformatics analysis and dual-luciferase reporter assay. Gain- and loss-of-function study also showed that when compared with miR-SC-DEX cells, miR-128-DEX cells showed significantly reduced SIRT1 expression and increased TG level, as well as elevated cellular levels of PPAR-γ, C/EBP-α and FAS and suppressed ATGL and HSL expression and enzyme activity. In contrast, miR-128-inhibitor-DEX cells precisely presented the opposite results. Collectively, these results indicate that miR-128 plays a role in the pathogenesis of glucocorticoid-related abnormal lipid accumulation via repressing SIRT1 expression, consequently, miR-128 inhibition may represent a novel potential therapeutic target in preventing DEX-induced abnormal lipid accumulation.

Regulatory mechanism of microRNA-128 in osteosarcoma tumorigenesis and evolution through targeting SASH1.

Osteosarcoma, which commonly occurs in young individuals, is a type of malignant tumor of growing bones. MicroRNAs (miRNAs) have been found to be involved in various cancer-related processes. In the present study, it was reported that miRNA-128 (miR-128) was overexpressed in pathological tissues from patients with osteosarcoma. The present study investigated the possible regulatory mechanism of miR-128 on the progression of osteosarcoma and offered a foundation for clinical therapeutics in osteosarcoma. First, the expressions levels of miR-128 and its target gene, SAM and SH3 domain-containing 1 (SASH1), were measured in tissues from patients with osteosarcoma, and their correlation with osteosarcoma in terms of the pathological level were examined. The effects of miR-128 on osteosarcoma cell proliferation and apoptosis were examined, and its regulation of the expression levels of SASH1 and associated proteins was analyzed. Subsequently, the association between SASH1 and miR-128 was evaluated using a dual luciferase gene reporter assay. Finally, an in vivo xenograft tumor mouse model of osteosarcoma was established to confirm the in vitro results. The results demonstrated a higher expression of miR-128 in pathological tissues, compared with that in normal tissues. From examining the patient osteosarcoma tissues, marked correlations were found between the expression of miR-128 and that of SASH1, particularly with tumor size, invasion depth, lymph node metastasis, and tumor-node-metastasis stage. Compared with the negative control group and blank control group, the results showed that the inhibition of miR-128 led to a lower cell proliferation rate and higher apoptotic rate in MG-63 cells (P<0.05). Additionally, the expression of B-cell lymphoma 2 (Bcl-2) was downregulated in the miR-128-inhibited group, compared with that in the control group, whereas the expression levels of SASH1, Bcl-2-associated X protein and caspase-3 were upregulated in the group with miR-128 inhibition (P<0.05). SASH1 was confirmed as a direct target of miR-128 using a dual luciferase gene reporter assay. Finally, the downregulation of miR-128 was found to induce tumor suppressive effects on xenograft tumor models of osteosarcoma in mice in vivo. The results of the present study suggested that miR-128 may regulate the tumorigenesis and evolution of osteosarcoma through targeting SASH1.

MicroRNA regulation of myogenic satellite cell proliferation and differentiation.

Myogenic satellite cells are stem cells responsible for muscle growth and regeneration. MicroRNAs (miRNAs) play significant roles in regulating numerous cellular processes. Two genes essential to satellite cell function are syndecan-4 and glypican-1. To determine if miRNAs influence myogenic satellite cell function, one miRNA predicted to bind syndecan-4 (miR-128) and two predicted to bind glypican-1 (miR-24 and miR-16) were inhibited in vitro by transfection of inhibitors targeting each miRNA. Inhibition of these miRNAs differentially affected the expression of syndecan-4, glypican-1, and myogenic regulatory factors myoD and myogenin. Inhibition of miR-16 reduced proliferation of satellite cells at 72h. Inhibition of miR-128 and miR-24 did not affect proliferation. Inhibition of miRNAs reduced differentiation of satellite cells into myotubes at 48 and 72h except for miR-16, which only affected differentiation at 72h. Inhibition of all three miRNAs decreased myotube width at 24h of differentiation and increased myotube width at 48h of differentiation. Inhibiting these miRNAs also increased the number of nuclei per myotube at 72h of differentiation. These data demonstrate individual miRNAs regulate genes essential for myogenic satellite cell proliferation and differentiation.

MiR128 up-regulation correlates with impaired amyloid β(1-42) degradation in monocytes from patients with sporadic Alzheimer's disease

Alzheimer's disease (AD), the most common form of dementia in elderly individuals, is characterized by neurofibrillary tangles, extracellular amyloid-β (Aβ) plaques and neuroinflammation. New evidence has shown that the lysosomal system might be a crossroad in which etiological factors in AD pathogenesis converge. This study shows that several lysosomal enzymes, including Cathepsin B, D, S, β-Galactosidase, α-Mannosidase, and β-Hexosaminidase, were less expressed in monocytes and lymphocytes from patients with a clinical diagnosis of AD dementia compared with cells from healthy controls. In vitro experiments of gain and loss of function suggest that down-regulation is a direct consequence of miR-128 up-regulation found in AD-related cells. The present study also demonstrates that miR-128 inhibition in monocytes from AD patients improves Aβ(1-42) degradation. These results could contribute to clarify the molecular mechanisms that affect the imbalanced Aβ production/clearance involved in the pathogenesis of AD.

Downregulation of miRNA‐128 sensitises breast cancer cell to chemodrugs by targeting Bax

miR-128 is more highly expressed in drug-resistant breast cancer samples than in drug-sensitive samples. We have confirmed that Bax is the target of miR-128 by negative post-transcriptional regulation. miR-128 and Bax were detected in the breast cancer cell line, MDA-MB-231, which was then transfected with miR-128 MIMIC (precursor of miR-128) or AMO (antisense-miR-128 oligonucleotides). After transfection, the chemosensitivity of MDA-MB-231 cell was up-regulated with increasing of Bax and inhibition of miR-128.