Relative Expression Of miR-21 Research Articles

Clinical role of serum microRNA-155 in early diagnosis and prognosis of septic patients with acute kidney injury.

Acute kidney injury (AKI) is a common complication in patients with sepsis, and early detection and timely treatment are crucial. This article aims to explore the clinical role of microRNA-155 (miR-155) in early diagnosis and prognosis evaluation of septic patients with acute kidney injury. We collected the blood samples of septic patients and measured the relative expression of serum miR-155 by RT-qPCR, and drew the receiver operating characteristic (ROC) curves to evaluate its early diagnosis for septic AKI. The relative expression of miR-155 in the septic AKI was significantly higher than that in the septic non-AKI, and increased with the aggravation of renal function damage. The ROC curve of miR-155 for the diagnosis of septic AKI was 1.91 (95% CI: 1.61-2.19). When the optimal cut-off value of miR-155 expression was 2.37, its sensitivity for diagnosing septic AKI was 91.12% (95% CI: 80.41-95.07%), and its specificity was 84.52% (95% CI: 71.74-89.36%). Furthermore, the severity of kidney injury, SOFA score, APACHE II score and miR-155 were the risk factors affecting the prognosis of septic patients with AKI. Serum miR-155 can be used as a novel biomarker for the early diagnosis of septic AKI, and also has important clinical value in the prognosis evaluation of septic patients with AKI.

SARS-CoV2 infection induce miR-155 expression and skewed Th17/Treg balance by changing SOCS1 level: A clinical study

BackgroundOne of the regulators in severe acute respiratory syndrome coronavirus2 (SARS-CoV2) infection is miRNAs. In COVID-19 patients, immunological responses to SARS-CoV2 infection may be impacted by miR-155, a miRNA associated to inflammation. Materials and methodsPeripheral blood mononuclear cells (PBMCs) of 50 confirmed COVID-19 patients /Healthy Controls (HCs) was isolated by Ficoll. The frequency of T helper 17 and regulatory T cells was analyzed by flowcytometry. The RNA was extracted from each sample and after synthesis of c-DNA, the relative expression of miR-155, suppressor of cytokine signaling (SOCS-1), Signal transducer and activator of transcription 3(STAT3), and Fork Head Box Protein 3 (FoxP3) was evaluated by real-time PCR. The protein level of STAT3, FoxP3 and RORγT in the isolated PBMCs measured by western blotting. The serum level of IL-10, TGF-β, IL-17 and IL21 was assessed by ELISA method. ResultsThe population of Th17 cells showed a significant rise, whereas Treg cells reduced in COVID-19 cases. The master transcription factor of Treg (FoxP3) and Th17 (RORγT) relative expression showed the same pattern as flowcytometry. STAT3 level of expression at RNA and protein level increased in COVID-19 cases. FOXP3 and SOCS-1 proteins were down-regulated. The relative expression of miR-155, up-regulated in PBMC of COVID-19 patients and revealed a negative correlation with SOCS-1. The serum cytokine profile showed a reduction in TGF-β, on the other hand an increase was seen in IL-17, IL-21 and IL-10 in COVID-19 cases toward control group. ConclusionBased on the studies conducted in this field, it can be suggested that Th17/Treg in covid-19 patients can be affected by miR-155 and it can be considered a valuable diagnostic and prognostic factor in this disease.

Experimental Study of Using Aloe Vera Gel to Treat Diabetic Chronic Cutaneous Ulcers in Bama Miniature Pigs

To investigate the efficacy, safety, and mechanism of topical application of aloe vera gel (AVG) to treat diabetic chronic cutaneous ulcers in Bama miniature pigs. The Bama miniature pig model of diabetic chronic skin wounds was constructed and the model pigs were randomly assigned to AVG daily administration group (AVG QD), aloe vera gel every-other-day administration group (AVG QOD), and diabetic control group (DC). A non-diabetic chronic skin wounds model pig was set as the non-diabetic control group (NDC). Treatment efficacy was evaluated based on the amount of time needed for complete healing of the wounds, healing rates, granulation growth rates, and skin histopathological changes. Safety was evaluated according to whether adverse reactions were observed. In addition, the dynamic changes of the relative expression levels of miR21, miR29a, miR126, miR146a, miR155, and miR210 in wound granulation tissues were examined. 1) Efficacy and safety: The amount of time needed for complete healing of the wounds was shorter in the NDC group than those of the three other groups, DC group, AVG QD group, and AVG QOD group (all P<0.05). The amount of time needed for complete healing of the wounds was shorter in the AVG QD group and AVG QOD group than that of DC group (all P<0.05). The amount of time needed for complete healing of the wounds was shorter in the AVG QOD group than that of AVG QD group (all P<0.05). No adverse reactions were detected in the whole process of AVG topical treatment. The granulation growth rate of NDC group was higher than those of DC group, AVG QD group, and AVG QOD group (all P<0.05). The wound healing rate of NDC group was higher than those of DC group, AVG QD group, and AVG QOD group (all P<0.05); the wound healing rate of AVG QOD group was higher than those of DC group and AVG QD group (all P<0.05). 2) Histopathology: The results of HE staining light microscopy showed that collagen fiber production increased, and that microvascular formation with slight inflammatory cell infiltration was observed in the dermal interstitium at the initial stage of wound healing after AVG treatment. One year of after complete healing, pathological examination results of wound healing skin showed that the epidermal keratinization was complete, that collagen was arranged neatly and orderly, and that many microvessels were found in the interstitium. The results of picric acid celestite scarlet staining showed that, after AVG treatment, type Ⅰ collagen mainly increased in the initial stage of wound healing, type Ⅲ collagen gradually increased when the wound healed completely, and the collagen was arranged neatly during the whole process. 3) The relative expression of microRNAs: The relative expression of miR21, miR126, and miR210 in NDC group, AVG QD group, and AVG QOD group were higher than that in DC group (all P<0.05). The relative expression of miR29a and miR155 in NDC group, AVG QD group, and AVG QOD group was lower than that in DC group (all P<0.05). The relative expression of miR146a in NDC group was higher than that in DC group ( P<0.05). AVG topical application can shorten the time needed for complete healing of diabetic chronic wounds in Bama minipigs. The wound healing speed of the alternate-day treatment group was faster than that of the daily treatment group. No adverse reactions were observed over the course of the treatment. The mechanism may be related to the up-regulation of the expressions of miR21, miR126, and miR210 and the down-regulation of miR29a and miR155 in wound granulation tissue.

Exosomes Derived from microRNA-21 Overexpressing Neural Progenitor Cells Prevent Hearing Loss from Ischemia-Reperfusion Injury in Mice via Inhibiting the Inflammatory Process in the Cochlea.

Both exosomes derived from neural progenitor cells (NPCs) can suppress inflammation. Whether exosomes derived from miR-21-transfected NPCs (miR-21-Exo) could be utilized to alleviate hearing loss is investigated. NPCs were transfected with lentiviral vectors overexpressing miR-21, and miR-21-Exo was purified. Morphology and exosome membrane markers were examined with nanoparticle tracking analysis, transmission electron microscopy, and Western blot. After incubation with different concentrations of miR-21-Exo, the viability of RAW 264.7 cells and the relative expressions of miR-21 and IL-10 were determined. The ischemia and reperfusion (I/R) model of C57BL/6 J mice was constructed, and the treatment benefit of miR-21-Exo was revealed by the auditory brainstem response (ABR) test. Immunofluorescence staining of caspase-3 and parvalbumin was used to detect apoptosis hair cells in the cochlea, and Western blot was utilized to detect the relative expressions of P53 and inflammatory cytokines in the cochlea. Isolated exosomes were confirmed by the size of 96 ± 25 nm, single membrane, and positive expression of CD9 and Tsg101. Upregulated miR-21 expression was detected in miR-21-transfected NPCs and miR-21-Exo. miR-21-Exo incubation demonstrated no cytotoxicity but upregulated miR-21 and IL-10 expressions in RAW 264.7 cells. The administration of miR-21-Exo inhibited the increased ABR threshold under 8, 16, and 32 kHz frequencies in cochlea-I/R injury mice and diminished the mean fluorescent intensity of caspase-3/parvalbumin. Moreover, miR-21-Exo treatment increased the IL-10 expression and prevented the increased TNF-α and IL-1β expressions in the cochlea of I/R mice both in mRNA and protein levels. Inner ear administration of miR-21-Exo effectively improved hearing damage caused by I/R.

Expression of miR-155 in Serum Exosomes in Children with Epilepsy and Its Diagnostic Value

Objective This study was designed to analyze the expression of miR-155 in serum exosomes in children with epilepsy and to explore its diagnostic value. Methods From March 2020 to March 2021, 43 hospitalized children with epilepsy admitted to the Department of Neurology of the hospital were included, and another 43 gender- and age-matched healthy children were randomly selected as the healthy control group during the same period. Then fasting serum samples of the two groups were collected to extract the exosomes. The morphology of the exosomes was evaluated under a transmission electron microscope, and the expression of specific protein markers on the surface was detected by Western Blot. In addition, the relative expression of miR-155 in serum exosomes in children with epilepsy with different courses of the disease and different degrees of abnormal electroencephalography (EEG) was compared, and the area under the receiver operating characteristic (ROC) curve (ROC-AUC) was used to evaluate the diagnostic value of miR-155. Results A higher relative expression level of miR-155 in serum exosomes was obtained in the epilepsy group, as compared to the healthy control group (P<0.05), and the relative expression of miR-155 in serum exosomes in children with epilepsy was correlated with the course of the disease and the degree of abnormal EEG (both P<0.05). In addition, the expression of miR-155 in serum exosomes showed high diagnostic efficiency for epilepsy (AUC = 0.813, P<0.05). Conclusion The expression of miR-155 in serum exosomes in children with epilepsy is up-regulated, and its level is related to the course of the disease and the degree of abnormal EEG, so miR-155 in serum exosomes may be used as a biomarker for the diagnosis and assessment of the severity of epilepsy.

The mir-21 Inhibition Enhanced HUVEC Cellular Viability during Hypoxia-Reoxygenation Injury by Regulating PDCD4.

The purpose of this study was to explore the clinical value of altered plasma mir-21 expression level as a biomarker for the severity of coronary artery disease (CAD) and its molecular impact on HUVEC cellular injuries. Angiographically validated 56 patients with single-vessel CAD disease, 92 patients with double-vessel CAD, 139 complex coronary artery stenosis patients, and 56 healthy individuals (n = 343) were enrolled in this study. The expressions of plasma mir-21 were evidently and progressively higher while PDCD4 levels were significantly and steadily lower in single-, dual-, and multivessel occluded CAD patients than in healthy participants (P < 0.001). The relative expressions of mir-21 in hypoxia-reoxygenation- (HR-) exposed HUVECs were markedly upregulated, but PDCD4 concentrations were obviously downregulated as compared with normal control cells (P < 0.001). Moreover, altered circulatory mir-21 expression levels were able to significantly differentiate single- (AUC 0.893), double- (AUC 0.914), and multivessel stenosis CAD (AUC 0.933) patients from healthy subjects. Besides, the plasma mir-21 expressions in elderly (66-85 years) groups were remarkably higher than those in younger aged (25-45 years) subjects. Caspase-3 and ROS expression levels were remarkably elevated, but cellular viability noticeably declined in HR-induced HUVECs than in normoxic cells (P < 0.001). In contrast, mir-21 inhibition markedly reduced caspase-3 activity and ROS concentrations while significantly ameliorating HUVEC cellular viability in HR conditions. PDCD4 expressions in HR-exposed HUVECs were prominently decreased whereas mir-21 inhibition significantly enhanced PDCD4 levels (P < 0.001). Upregulated plasma mir-21 can be a valuable clinical biomarker for the detection of the severity of coronary artery stenosis patients. Elevated circulatory mir-21 concentrations have a positive correlation with aging. Inhibitory mir-21 evidently increased HUVEC cellular viability through upregulation of targeting PDCD4 and recommended a newer possible therapeutic molecule for the management of CAD patients.

MicroRNA155 May Be an Important Part of Human Papilloma Virus-Related Cancers

Background: Micro-RNAs (miRs) play several roles during infections with viruses. Therefore, the roles of miR21 and mir155 in the induction of the viral-related cancers have been the focus of attention in several studies. High risk human papilloma viruses (HPVs) are the main factors negatively contributing to the induction of HPV-related cancers. This study aimed to evaluate the expression of miR21 and mir155 in the patients with HPV-high risk genotypes in order to explore the roles of the miRs in the induction of HPV-related cancers. Methods: In this study, 40 women infected with the high-risk HPV genotypes as well as 40 healthy controls were examined regarding the relative expression of miR21 and mir155 by adopting real-time polymerase chain reaction (PCR) technique. U6 was used for data normalization of miRNAs. Results: Relative expressions of both miR21 and miR155 were significantly higher in the HPV-infected patients compared to those in non-infected women. Conclusions: It was concluded that miR21 and miR155 may have played key roles in the induction of HPV-related cancers among Iranian patients.

Relationship between Polymorphism of miR-155 and Its Target Gene MyD88 and Clinicopathological Features of Diffuse Large B-cell Lymphoma

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). 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. 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； χ2=10.681、8.599、7.251、23.008；P<0.05). The survival of DLBCL patients with low miR-155 expression was significantly better than that with high miR-155 expression (P<0.05). The survival of wild-type DLBCL patients with MyD88 L265P locus was significantly better than that of mutant DLBCL patients (P<0.05). There was a significant positive correlation between the relative expression of miR-155 and the expression of MyD88 protein (r=0.428, P=0.000). 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.

MicroRNA-21 ameliorates the impairment of autophagy in palatal wound healing.

Fibroblast injury and autophagy dysfunction have been shown to contribute to the persistence of oral wounds. Recently, microRNAs have emerged as vital regulators and fine tuners of various pathophysiological cellular processes that influence the wound healing process. This study explored the biological function and regulatory mechanism of miRNA-21 (miR-21) in the healing of oral wounds by interfering with autophagy. Healthy gingival cells derived from wild-type (WT) and from miR-21KO mice were characterized by immunocytofluorescence, and changes in wound healing were subsequently assessed using an in vitro scratch wound healing assay. The roles of critical proteins required for autophagy, autophagy related 5 (ATG5) and Bcl-2 interacting coiled-coil protein 1 (Beclin1) were evaluated by immunohistochemistry. Human gingival fibroblasts (HGFs) were transfected with a miR-21 mimic and a miR-negative control, and the relative expression of miR-21, ATG5, Beclin1 and LC3-I/II was characterized by qRT-PCR and Western blot. Pathological changes were observed in a palatal wound healing model using WT and miR-21KO mice. Immunohistochemistry was used to examine extracellular matrix (ECM) proteins and autophagy markers. Cell migration was delayed in gingiva-derived mesenchymal stem cells (GMSCs) from miR-21KO mice compared with WT mice. The expression of ATG5 and Beclin1 was significantly up-regulated in miR-21KO gingiva. Transfection of a miR-21 mimic into HGFs inhibited autophagy and up-regulated miR-21 expression. Knockdown of miR-21 suppressed the expression of fibronectin and CTGF, enhanced the autophagy effect of fibroblasts, suggesting that autophagy is involved in miR-21 regulated palatal wound healing. Taken together, these results suggest that miR-21 promotes oral wound healing by increasing ECM production through the inhibition of autophagy and facilitates clinical management of wound healing.

Evaluation of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer.

To evaluate the clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer (BC). 129 patients with BC and 50 patients with benign breast lesions were selected. They were scanned by color Doppler ultrasound on the breast. The relative expression of miR-21 and miR-27a in plasma was detected by real-time fluorescence quantitative PCR. The clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC was analyzed retrospectively. Two-dimensional ultrasound images of BC showed irregular shape, unclear boundary, burr-like edge, uneven internal echo, weakened rear echo, microcalcification, aspect ratio > 1 and axillary lymph node metastasis. The grade of blood flow signal was improved, mainly in grade II and above, which was statistically different from that of benign control group (P < 0.001). The relative expression of plasma miR-21 and miR-27a in breast cancer group was significantly higher than that in benign control group (P < 0.001). The relative expressions of plasma miR-21 and miR-27a in breast cancer group were significantly correlated with histological grade, clinical stage, lymph node metastasis and postoperation. The relative expression levels of miR-21 and miR-27a in plasma of patients with low histological grade (grade III), high clinical stage (stage III, IV) and lymph node metastasis before operation were significantly higher than those in patients with high histological grade (grade I, II), low clinical stage (stage I, II) and no lymph node metastasis after operation (P < 0.01). The sensitivity, accuracy and negative predictive value of ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC were significantly higher than those of each single examination. Color Doppler ultrasound combined with plasma miR-21 and miR-27a can significantly improve the diagnostic efficiency of BC, which is beneficial to early diagnosis and early clinical intervention.

Upregulation of microRNA-155 Enhanced Migration and Function of Dendritic Cells in Three-dimensional Breast Cancer Microenvironment

ABSTRACT Background: Dendritic cells (DCs) play an essential role in the induction and regulation of immune responses, including the activation of effector T lymphocytes for the eradication of cancers. However, the tumor microenvironment (TME) often leads to DCs dysfunction due to their immature state. MicroRNA-155 (miR-155) has emerged as a typical multifunctional gene regulator associated with immune system development and immune cell activation and differentiation. Methods: In this study, a three-dimensional TME model that closely mimics the microenvironment of breast cancer was prepared. MiR-155 overexpression and control vectors were constructed using lentivirus. The relative expression of miR-155 was determined by qRT-PCR. Cell viability, antigen uptake and cell surface marker expression were analyzed by live-dead staining and flow cytometry. The migration ability of bone marrow-derived DCs (BMDCs) was qualified by transwell assay. A mixed lymphocyte culture assay was used to assess T cell-specific proliferation. Cytokine levels were determined by ELISA. Results: We found that the expression of miR-155 in DCs was inhibited by the TME. Furthermore, upregulation of miR-155 enhanced the migration ability, uptake of antigen and elevated the expression of the mature DCs markers CD80 and MHCII. More importantly, overexpression of miR-155 in DCs significantly induced T cell proliferation and IFN-γ and IL-2 secretion. Conclusion: MiR-155 is a potential molecular regulator that may improve the efficacy of DCs-based tumor immunotherapy.

Lidocaine promotes autophagy of SH-SY5Y cells through inhibiting PI3K/AKT/mTOR pathway by upregulating miR-145.

Lidocaine is one of the most common local anesthetics (LA) used in clinical practice and it is neurotoxic. Recent studies suggested that LA, including lidocaine, could exert protective effect over neurotoxicity by promoting autophagy. However, the underlying mechanism was not sufficiently elucidated. This study aimed to explore the mechanism behind. Human neuroblastoma cell line SH-SY5Y was used throughout the whole study. The effect of lidocaine on viability, toxicity of SH-SY5Y cells were analyzed by MTT and lactate dehydrogenase (LDH) assays, respectively. The relative expression of miR-145 was assessed by quantitative reverse transcription-polymerase chain reaction. The impact which lidocaine brought on PI3K/AKT/mTOR pathway and autophagy-related proteins were examined by the western blot assay. LC3B was assessed by immunofluorescence staining. The interaction between miR-145 and AKT3 was conducted by the dual-luciferase reporting assay. Lidocaine inhibited viability of SH-SY5Y cells in a time and dose dependent manner and enhanced the release of LDH in SH-SY5Y cells. Furthermore, the expression of miR-145 and autophagy were enhanced by lidocaine. Transfection with miR-145 inhibitor inhibited the release of LDH and autophagy. miR-145 targeted AKT3 to inhibit PI3K/AKT/mTOR pathway. Finally, lidocaine inactivated PI3K/AKT/mTOR pathways via upregulation of miR-145, and it subsequently promoted autophagy of SH-SY5Y cells. However, silence of miR-145 could reverse the promotion of the autophagy of SH-SY5Y cells. Our results showed that lidocaine promoted autophagy of nerve cells via regulating miR-145 expression and further inactivation of PI3K/AKT/mTOR signaling pathway.

Resveratrol promoted the M2 polarization of microglia and reduced neuroinflammation after cerebral ischemia by inhibiting miR-155

Purpose: Neuroinflammation was demonstrated to play an important role in the brain injury induced by cerebral ischemia, which was mainly mediated by microglia. MicroRNA-155 (MiR-155) was reported to promote the M1 polarization of microglia and increase neuroinflammation. Resveratrol was identified to have the ability to promote the M2 polarization of microglia and reduce inflammation. Whether resveratrol can promote the M2 polarization of microglia and further inhibit neuroinflammation after cerebral ischemia, and its correlation with miR-155 is unclear. To clarify this, we conducted this study to explore the potential of resveratrol as an effective strategy to treat cerebral ischemia induced neuroinflammation.Materials and methods:The cerebral ischemia mouse model was first constructed by middle cerebral artery occlusion (MCAO). Then resveratrol was intraperitoneally injected at 0 h, 8 h and 18 h after cerebral ischemia. Subsequently, the relative expression of miR-155 and the signature genes of M1 and M2 microglia in injured brain were measured by RT-PCR, and the concentration of pro-inflammatory and anti-inflammatory cytokines were detected by ELISA. Further, the in vitro experiments were also conducted to explore the effect of resveratrol on the inflammation mediated by LPS activated BV2 microglia.Results: Results indicated that the relative expression of miR-155 in ischemia brain and activated BV2 microglia was elevated, while resveratrol reduced the expression of miR-155. Resveratrol promoted the M2 polarization of microglia and reduced neuroinflammation in injured brain and activated BV2 microglia.Conclusions: In conclusion, this research indicated that resveratrol promoted the M2 polarization of microglia and reduced neuroinflammation after cerebral ischemia by inhibiting miR-155.

Regulation of JAK/STAT signal pathway by miR-21 in the pathogenesis of juvenile idiopathic arthritis

BackgroundOverexpression of the components of the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signalling pathway is the key factor of the pathogenic mechanisms underlying systemic juvenile idiopathic arthritis (sJIA). The study aims to investigate the association between miR-21 and the JAK/STAT signal pathway in JIA.MethodsTotal RNA was extracted from peripheral blood mononuclear cells (PBMCs) in active JIA patients. The relative expressions of miR-21, STAT3 and suppressor of cytokine signalling 3 in PBMCs were measured by real-time polymerase chain reaction and their expressions were measured by western blotting and dual-luciferase reported assay. Rheumatoid arthritis fibroblast-like synovial cell (RASF) was stimulated to become to osteoclasts using macrophage colony-stimulating factor (M-CSF) and factors that can impact on their differentiation ability were identified through the transfection of LV3-miR-21. The expression of STAT3/p-STAT3 was measured by western blot, and the levels of interleukin (IL)-17A, p65, matrix metalloproteinases (MMP)-3, MMP-4 and receptor activator of nuclear factor-κB after the LV3-miR-21 transfection were tested by enzyme-linked immunosorbent assay. Finally, the miR-21 targeted STAT3 gene was detected by the dual-luciferase reported assay.ResultsThe expression of miR-21 was significantly lower in JIA patients than in healthy control (P < 0.05). The level of STAT3 was increased in PBMCs of JIA group compared with control group (P < 0.05). Furthermore, the expression levels of miR-21 in sJIA and polyarticular JIA groups were negatively correlated with STAT3 (r = − 0.5854/r = − 0.6134, P < 0.05). The expression of STAT3 changed little in PBMCS after the stimulation of IL-6 and not in RASFs with transfection of LV3-miR-21. The expression of p-STAT3 decreased after the stimulation of IL-6 in RASFs transfected by LV3-miR-21 (P < 0.05). RASFs were induced into osteoclasts using M-CSF. The number of osteoclasts as determined by tartrate-resistant acid phosphatase staining was significantly lower in group miR-21 mimics as compared with the negative control group (P < 0.05).ConclusionsWe showed that expression of miR-21 was significantly lower in JIA patients compared with healthy control. MiR-21 might affect the JAK/STAT signal pathway by suppressing the expression of STAT3 and phosphorylation of STAT3. MiR-21 could inhibit the production of osteoclasts induced from RASFs by M-CSF.

RETRACTED ARTICLE: Lidocaine inhibits growth, migration and invasion of gastric carcinoma cells by up-regulation of miR-145

BackgroundGastric cancer receives considerable attention not only because it is the most common cancer all through the world, but also because it’s on the top third leading reason for cancer-related death. Lidocaine is a well-documented local anesthetic that has been reported to suppress cancer development. The study explored the effects of lidocaine on the growth, migration and invasion of the gastric carcinoma cell line MKN45 and the mechanism behind.MethodsThe effect of lidocaine on viability, proliferation and apoptosis of MKN45 cells were analyzed by Cell Counting Kit-8 assay, BrdU staining assay and flow cytometry, respectively. Moreover, cell migration and invasion were both examined by Transwell assay. The expression of apoptosis-, migration-, and invasion-related proteins were detected by western blot. The relative expression of miR-145 was determined by qRT-PCR. Moreover, the impact which lidocaine brought on MEK/ERK and NF-κB pathways were examined by western blot.ResultsLidocaine inhibited viability, proliferation, migration, and invasion of MKN45 cells, while enhanced apoptosis. Moreover, miR-145 expression was enhanced by lidocaine; and transfection with miR-145 inhibitor increased cell viability, proliferation, migration, and invasion, but inhibited apoptosis. The up-regulation of miR-145 was partly contributed to the inhibitory effect of lidocaine on gastric cancer cell line MKN45. Finally, lidocaine inactivated MEK/ERK and NF-κB pathways via up-regulation of miR-145.ConclusionsOur results suggested that lidocaine decreased growth, migration and invasion of MKN45 cells via regulating miR-145 expression and further inactivation of MEK/ERK and NF-κB signaling pathways.

Lidocaine alleviates cytotoxicity-resistance in lung cancer A549/DDP cells via down-regulation of miR-21.

Lidocaine (Lido) is a commonly used local anesthetic, which has been reported in various types of cells. However, the effects of Lido on lung cancer cells remain not understood. The study aimed to investigate the underlying mechanisms of Lido in the cisplatin resistance of A549/DDP cells. Different concentrations of cisplatin (0-320µM) were used to stimulate A549 and A549/DDP cells, and cell viability and apoptosis were examined. To investigate the effect of Lido on A549/DDP cells, the optimum concentration of Lido was selected to treat A549/DDP cells, and cell viability, apoptosis, migration and invasion were then detected. The relative expression of miR-21 in A549/DDP cells or in Lido-treated A549/DDP cells was analyzed by RT-qPCR. MiR-21 mimic, inhibitor and its control were transfected into A549/DDP cells to explore the regulatory effect of miR-21 on the cisplatin resistance in A549 or A549/DDP cells. The effects of miR-21 on PTEN/PI3K/AKT and PDCD4/JNK pathways were detected by western blot. The cisplatin resistance of A549/DDP cells was higher than that of A549 cells. Lido significantly suppressed cell viability, induced apoptosis, and inhibited cell migration and invasion in A549/DDP cells. Additionally, miR-21 expression in A549/DDP was higher than that in A549 cells, and Lido significantly down-regulated miR-21 expression in A549/DDP cells. MmiR-21 inhibition exhibited the same effects as Lido on the cisplatin resistance of A549/DDP cells. Further, miR-21 suppression regulated PTEN/PI3K/AKT and PDCD4/JNK pathways in A549/DDP cells. These findings indicated that Lido alleviated the cytotoxicity resistance of A549/DDP cells via down-regulation of miR-21.

Circulating miR-21 and miR-155 as potential noninvasive biomarkers in Iranian Azeri patients with breast carcinoma.

Breast cancer (BC) is the most common cause of cancer-related mortality among women. Despite recent advances in diagnosis and prognosis of breast carcinomas, noninvasive biomarkers have been poorly identified. We evaluated the biomarker potential of miR-21 and miR-155 in tissue and plasma specimens of Iranian Azeri patients. Tumor specimens, paired nontumoral adjacent tissues, and matched plasma samples were collected from a number of thirty Iranian Azeri women with breast carcinoma. Plasma of healthy women was used as the control. The relative expression of miR-21 and miR-155 was measured by real-time polymerase chain reaction. Our data revealed that the expression levels of miR-21 and miR-155 in tumor tissues are significantly higher than paired nontumoral adjacent specimens (P < 0.05). Furthermore, receiver operating characteristic (ROC) curve analysis of samples showed the area under the ROC curve of 0.81 for miR-21 and area of 0.83 for miR-155. In addition, statistical analysis showed that miR-21 and miR-155 RNAs are significantly detected in the plasma of BC patients compared to healthy specimens (P < 0.05). Circulating miRNAs yielded area under the ROC curve of 0.99 for miR-21 and 0.92 for miR-155. Our data showed that miR-21 and miR-155 oncomiRs can be considered as noninvasive biomarkers for monitoring breast carcinomas. However, further investigations are needed to confirm the use of these noncoding RNAs in pathology.

Expression of miRNA-21 in mucosa-associated lymphoid tissue lymphomas and its relationship with CHOP regimen

Objective To investigate the expression of miRNA (miR-21) in mucosa-associated lymphoid tissue(MALT) lymphomas and its relationship with the effects in CHOP regimen. Methods Lymph gland tissues and preoperative peripheral blood of 52 patients pathologically diagnosed with MALT lymphoma in Department of Hematology of Affiliated Hospital of Hebei University of Engineering from January 2015 to December 2017 were collected; meanwhile, 10 tissues from patients with lymphadenitis and 20 peripheral serum from healthy examination patients were also collected. Quantitative real-time polymerase chain reaction (qPCR) was used to compare the lymph gland tissues in MALT lymphoma patients and lymphadenitis patients, and the expression of miR-21 in peripheral serum of MALT lymphoma and healthy people. The selected 20 cases of MALT lymphoma were given CHOP regimen treatment for 6 cycles, and then the efficiency and inefficiency were compared; at the same time, the expression of miR-21 in peripheral serum of the patients in the effective group was detected. Results The relative expression of miR-21 of lymph node tissues in lymphadenitis patients and MALT lymphoma patients was 1.03±0.12 and 4.53±0.73 respectively, and the relative expression of miR-21 in preoperative peripheral serum of healthy people and MALT lymphoma patients was 0.83±0.04 and 3.87±0.21 respectively, and the differences were statistically significant (P= 0.047, P= 0.044). After 6 cycles of CHOP regimen, the total effect rate of MALT lymphoma was 70% (14/20), and the relative expression of miR-21 in preoperative and postoperative peripheral serum for the patients who obtained the effective results after CHOP regimen was 3.95±0.08 and 1.62±0.41, and the differences were statistically significant (P= 0.035). Conclusions The expression of miR-21 has a certain correlation with the occurrence and development of MALT lymphoma. Furthermore, the serum miR-21 expression may be related to the effect of CHOP regimen chemotherapy in MALT lymphoma. Key words: Mucosa associated-lymphoid tissue lymphoma; miRNA-21; CHOP regimen; Treatment outcome

Targeting the polarization of tumor-associated macrophages and modulating mir-155 expression might be a new approach to treat diffuse large B-cell lymphoma of the elderly.

Aging immune deterioration and Epstein-Barr (EBV) intrinsic mechanisms play an essential role in EBV-positive diffuse large B-cell lymphoma (DLBCL) of the elderly (EBV + DLBCLe) pathogenesis, through the expression of viral proteins, interaction with host molecules and epigenetic regulation, such as miR-155, required for induction of M1 phenotype of macrophages. This study aims to evaluate the relationship between macrophage polarization pattern in the tumor microenvironment and relative expression of miR-155 in EBV + DLBCLe and EBV-negative DLBCL patients. We studied 28 EBV + DLBCLe and 65 EBV-negative DLBCL patients. Tumor-associated macrophages (TAM) were evaluated by expression of CD68, CD163 and CD163/CD68 ratio (degree of M2 polarization), using tissue microarray. RNA was extracted from paraffin-embedded tumor samples for miR-155 relative expression study. We found a significantly higher CD163/CD68 ratio in EBV + DLBCLe compared to EBV-negative DLBCL. In EBV-negative DLBCL, CD163/CD68 ratio was higher among advanced-staged/high-tumor burden disease and overexpression of miR-155 was associated with decreased polarization to the M2 phenotype of macrophages. The opposite was observed in EBV + DLBCLe patients: we found a positive association between miR-155 relative expression and CD163/CD68 ratio, which was not significant after outlier exclusion. We believe that the higher CD163/CD68 ratio in this group is probably due to the presence of the EBV since it directly affects macrophage polarization towards M2 phenotype through cytokine secretion in the tumor microenvironment. Therapeutic strategies modulating miR-155 expression or preventing immuno-regulatory and pro-tumor macrophage polarization could be adjuvants in EBV + DLBCLe therapy since this entity has a rich infiltration of M2 macrophages in its tumor microenvironment.

Abnormal Expression of miR-21 in Kidney Tissue of Dogs With X-Linked Hereditary Nephropathy: A Canine Model of Chronic Kidney Disease.

MicroRNAs (miRNAs) are a group of small noncoding RNAs that act as regulators of posttranslational gene/protein expression and are known to play a key role in physiological and pathological processes. The objective of our study was to compare expression of miR-21 in renal tissue from dogs affected with chronic kidney disease (CKD) caused by X-linked hereditary nephropathy (XLHN), a disease equivalent to human Alport syndrome, to that from unaffected dogs. Additionally, we sought to characterize changes in relative mRNA expression of various genes associated with miR-21 function. miRNA was isolated from kidney tissue collected from both affected dogs and unaffected, age-matched littermates at defined milestones of disease progression, including end-stage renal disease (ESRD). Additionally, autopsy samples from affected dogs at ESRD and corresponding unaffected dogs were evaluated. Samples were scored based on histological changes, and relative expression of miR-21 and kidney disease-related genes was determined using quantitative real-time polymerase chain reaction. In affected dogs, significant upregulation of kidney miR-21 was first detected at the milestone corresponding with increased serum creatinine. Furthermore, miR-21 expression correlated significantly with urine protein: urine creatinine ratio, serum creatinine concentration, glomerular filtration rate, and histologic lesions (glomerular damage, tubular damage, chronic inflammation, and fibrosis). At end-stage disease, COL1A1, TGFB1 and its receptor, TGFB2, and Serpine1 were upregulated, while PPARA, PPARGC1A, ACADM, SOD1, and EGF were downregulated. In conclusion, miR-21 is abnormally upregulated in the kidneys of dogs with CKD caused by XLHN, which may play an important pathologic role in the progression of disease by dysregulating multiple pathways.