MiR-222 inhibition alleviates Staphylococcal Enterotoxin B-induced inflammatory acute lung injury by targeting Foxo3

Acute lung injury (ALI) is a common acute and severe disease in clinical practice. Staphylococcal EnterotoxinB (SEB) is a superantigen that can cause inflammatory ALI. MiR-222 has been demonstrated to be upregulatedin SEB-induced inflammatory ALI, but its exact roles and functions remain ill-defined. In this study, SEBexposure led to inflammatory ALI and high expression of miR-222 in model mice and lung infiltratingmononuclear cells, but the inflammatory response and high expression of miR-222 were restored in miR-222-/-mice. Moreover, we investigated the roles of miR-222 in vitro and observed that the concentrations ofinflammatory cytokines and the expression of miR-222 were all elevated in SEB-activated splenocytes andmiR-222 inhibition reversed the effects. Foxo3 was confirmed as a direct target of miR-222. Interestingly, SEBexposure led to a decrease of Foxo3 expression, and Foxo3 knockdown partially reversed the promotion ofFoxo3 and the inhibition of inflammatory cytokines induced by miR-222 inhibitor in SEB-activated splenocytes. Our data indicated that miR-222 inhibition could alleviate SEB-induced inflammatory ALI by directlytargeting Foxo3, shedding light on the potential therapeutic of miR-222 for SEB-induced inflammation in thelung.

Inhibition of miRNA-222-3p Relieves Staphylococcal Enterotoxin B-Induced Liver Inflammatory Injury by Upregulating Suppressors of Cytokine Signaling 1

PURPOSE: Staphylococcal enterotoxin B (SEB) has been well-documented to induce liver injury. miRNA-222-3p (miR-222-3p) was implicated in SEB-induced lung injury and several liver injuries. This study aimed to explore the role of miR-222-3p in SEB-induced liver injury. MATERIALS AND METHODS: Expression of miR-222-3p and suppressors of cytokine signaling 1 (SOCS1) was detected using real-time quantitative PCR and western blot. Liver injury was determined by levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and inflammatory cytokines, numbers of infiltrating mononuclear cells using AST/ALT assay kit, enzyme-linked immunosorbent assay (ELISA), and hematoxylin-eosin staining, respectively. Target binding between miR-222-3p and SOCS1 was predicted on targetScan software, and confirmed by luciferase reporter assay. RESULTS: SEB induced liver injury in D-galactosamine (D-gal)-sensitized mice, as demonstrated by increased serum levels of AST and ALT, elevated release of interferon-gamma (INF-γ), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-2, and promoted infiltrating immune cells into liver. Expression of miR-222-3p was dramatically upregulated, and SOCS1 was downregulated in SEB-induced liver injury both in mice and splenocytes. Moreover, miR-222-3p knockout (KO) mice exhibited alleviated liver injury accompanied with SOCS1 upregulation. Besides, splenocytes under SEB challenge released less INF-γ, TNF-α, IL-6, and IL-2 during miR-222-3p knockdown. Mechanically, SOCS1 was targeted and downregulated by miR-222-3p. Upregulation of SOCS1 attenuated INF-γ, TNF-α, IL-6, and IL-2 release in SEB-induced splenocytes; downregulation of SOCS1 could block the suppressive role of miR-222-3p knockdown in SEB-induced splenocytes. CONCLUSION: Inhibition of miR-222-3p relieves SEB-induced liver inflammatory injury by upregulating SOCS1, thereby providing the first evidence of miR-222-3p in SEB-induced liver injury.

Expression of miR-21, miR-221, and miR-222 in exosomes of CAL27 tongue squamous cell carcinoma cells / 口腔疾病防治

Objective@# To assess the expression of miR-21, miR-221, and miR-222 in exosomes of CAL27 tongue squamous cell carcinoma cells.@*Methods @# CAL27 tongue squamous cell carcinoma cells and normal human oral keratinocytes (HOKs) were cultured, and then, the cultured supernatant was collected to separate the exosomes. Exosomes were detected by electron microscopy, and the expression levels of miR-21, miR-221, and miR-222 in the exosomes of tongue cancer cells were measured by qRT-PCR. @*Results@#Exosomes existed in the cultured supernatants of CAL27 cells and HOKs. Additionally, the expression levels of miR-21, miR-221, and miR-222 in the exosomes of CAL27 cells were significantly enhanced compared with those in the HOK exosomes (P < 0.05). @*Conclusion @#The expression levels of miR-21, miR-221, and miR-222 were markedly enhanced in the exosomes of CAL27 tongue squamous cell carcinoma cells.

MicroRNA-222 Expression as a Predictive Marker for Tumor Progression in Hormone Receptor-Positive Breast Cancer / 한국유방암학회지

PURPOSE: The microRNA-221/222 (miR-221/222) gene cluster has been reported to be associated with the promotion of epithelial-mesenchymal transition (EMT), downregulation of estrogen receptor-α, and tamoxifen resistance in breast cancer. We studied the expression of miR-222 in human breast cancer samples to analyze its relationship with clinicopathologic features of the tumor, including estrogen receptor status, expression of EMT markers, and clinical outcomes. METHODS: Quantitative real-time polymerase chain reaction was performed to detect the expression of miR-222 in 197 invasive breast cancers. Expression of EMT markers (vimentin, smooth muscle actin, osteonectin, N-cadherin, and E-cadherin) was evaluated using immunohistochemistry. RESULTS: High miR-222 levels were associated with high T stage, high histologic grade, high Ki-67 proliferation index, and HER2 gene amplification. Its expression was significantly higher in the luminal B and human epidermal growth factor receptor 2-positive (HER2+) subtypes than in the luminal A and triple-negative subtypes. In the hormone receptor-positive subgroup, there was a significant negative correlation between miR-222 and estrogen receptor expression, and miR-222 expression was associated with EMT marker expression. In the group as a whole, high miR-222 expression was not associated with clinical outcome. However, subgroup analyses by hormone receptor status revealed that high miR-222 expression was a poor prognostic factor in the hormone receptor-positive subgroup, but not in the hormone receptor-negative subgroup. CONCLUSION: This study showed that miR-222 is associated with down-regulation of the estrogen receptor, EMT, and tumor progression in hormone receptor-positive breast cancer, indicating that miR-222 might be associated with endocrine therapy resistance and poor clinical outcome in hormone receptor-positive breast cancer.

Expression of miR-2 2 1 and miR-2 2 2 in Papillary Thyroid Carcinoma Tissues and Its Clinical Significance / 现代检验医学杂志

Objective To explore the expressions of miR-221 and miR-222 in papillary thyroid carcinoma (PTC),and relativi-ties with clinical pathological features.Methods Samples from patients of PCT (43 cases),nodular golter(21 cases),and para-carcinoma thyroid tissues(14 cases),78 cases in total (from 06/2015 to 05/2016,Shaanxi Provincial People’s Hospital) were collected.Real time-PCR tests were carried out,then analyzed in relation to clinical pathology features,and statistical a-nalysis was used to evaluate the results.Results The expressions of miR-221 and miR-222 were significantly higher in PTC (11.54±3.37,10.67±2.45)than in nodular golter (3.21±1.12,2.89±1.23)and normal thyroid tissue (2.02±0.76, 1.98±0.34)(t=3.62,3.25;3.27,3.01,all P0.05),and no differences were found in nodular golter and in normal thyroid tissue (t=0.91, 0.79,P>0.05).Conclusion miR-221 and miR-222 could be considered as a specific molecular marker of PTC,may play an important role in the diagnosis and treatment on PTC.

Effect and mechanism of exercise-induced miR-222 in the protection of myocardial damage in diabetic mice / 中华内分泌代谢杂志

Objective To investigate the protective effects of aerobic exercise and miR-222 expression in myocardium of diabetic mice. Methods C57BL/ 6 mice were divided into 4 groups: normal non-exercise group (SC), normal exercise group(EC), non-exercise diabetic group(SD), and exercise diabetic group(ED). After the diabetic model was established successfully, EC and ED underwent a swimming training for 5 weeks. By the end of the experiment, light microscope was used to observe the pathological changes of heart, RT-PCR for myocardial miR-222 expression, and Western blot for phosphatase and tensin homolog deleted on chromosome ten ( PTEN ), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt) protein expressions in myocardial tissue. Results (1) Under the light microscope, the diabetic mice had a significant change in myocardial structure, with great disorder in the cell arrangement. After exercise intervention, the lesion was alleviated. (2) MiR-222 expression was increased in the myocardium of normal mice and DM mice after exercise (all P<0. 05); (3) Compared with SC group, PTEN expression was increased and PI3K/ Akt expressions were inhibited in myocardium of diabetic mice(all P <0. 05). After exercise intervention, the expression of PTEN reduced( P < 0. 05) and PI3K/ Akt pathway was reactivated in myocardium of diabetic mice (all P<0. 05). Conclusion Exercise intervention may protect the myocardium under high glucose via inducing miR-222 and activating PI3K/ Akt signaling pathway.

The expression of miR-221 and miR-222 in non-small cell lung cancer and their significances / 肿瘤研究与临床

Objective To explore the expression of miR-221/222 in non-small cell lung cancer (NSCLC) and its correlation with clinical pathological parameters. Methods The clinical pathological data and formalin fixed-paraffin embedded (FFPE) tissues of 55 NSCLC patients and 10 benign lesion patients who underwent surgery in the First Affiliated Hospital of Nanhua University from February 2012 to May 2014 were collected and followed. The relationship between miR-221/222 expression detected by real-time PCR and clinical pathological parameters and progression-free survival (PFS) were analyzed. The differential survival between the high expression group and the low expression group of miR-221/222 were compared. A Cox proportional hazard regression model was utilized to examine the prognostic factors of NSCLC. Results The expression level of miR-221/222 was significantly higher in tumor tissues than that in corresponding benign lesion tissues (Fold change=3.52, P=0.000;Fold change=2.01, P=0.000). There was a negative correlation between miR-221/222 expression and pathological grades (r=-0.732, P=0.000;r=-0.451, P=0.001). The relative expression of miR-221 showed a positive correlation with miR-222 (r=0.376, P=0.000). Patients with higher levels of miR-221/222 were closely associated with a shorter PFS (miR-221: 55.43 weeks vs. 81.29 weeks, P=0.028; miR-222: 45.00 weeks vs. 87.04 weeks, P=0.008). Finally, multivariate analysis demonstrated that miR-222 expression was independently associated with poor PFS (RR=2.808, P=0.033). Conclusions miR-221/222 is highly expressed in NSCLC tumor tissues with a positive correlation. A negative correlation is observed between the expression of miR-221/222 and tumor differentiation. The potential high expression of miR-221/222 is considered as tumor biomarkers for the prognosis of NSCLC patients.

Clinical Observation of Edaravone Combined with Butyl Phthalide in the Treatment of Acute Ischemic Stroke / 中国药房

OBJECTIVE:To observe clinical efficacy and safety of edaravone combined with butyl phthalide in the treatment of acute ischemic stroke (AIS). METHODS:258 AIS patients were randomly divided into observation group and control group, with 129 cases in each group. Both groups were given routine treatment as antiplatelet,improving microcirculation,controlling blood pressure,lowering blood glucose,regulating blood lipid,keeping plaque stable,nourishing brain cells. Control group was additionally given Butyl phthalide capsules orally,200 mg,tid. Observation group was additionally given Edaravone injection 30 mg added into Sodium chloride injection 100 ml,ivgtt,bid,on the basis of control group. Both groups continuously received 14 days of treatment. The serum inflammatory factor,miR-222 and neurologic impairment score of 2 groups were observed before treatment,7,14 d after treatment. Clinical efficacies and the occurrence of ADR were compared between 2 groups. RESULTS:There was no statistical significance in the serum inflammatory factor,miR-222 and neurologic impairment score between 2 groups before treatment(P>0.05). The serum inflammatory factor and neurologic impairment score of 2 groups were decreased significant-ly 7,14 d after treatment,while serum levels of miR-222 were increased significantly;the observation group was significantly bet-ter than the control group,with statistical significance(P<0.05). Total effective rate of observation group was 92.2%,which was significantly higher than that of control group (69.8%),with statistical significance (P<0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:Edaravone combined with butyl phthalide is effective in the treatment of AIS,and can significantly de-crease serum inflammatory factor level,promote the expression of miR-222 and improve neurologic function with good safety.

miR-222 promotes retinoblastoma cell proliferation and invasion by targeting RB1 / 中国癌症杂志

Background and purpose:A large number of studies have showed that retinoblastoma gene 1 (RB1) can inhibit the occurrence and development of many tumors, including neuroblastoma, small cell lung cancer, osteosar-coma, pancreatic cancer, breast cancer and so on. RB1 is also closely related to the regulation of cell cycle, differentia-tion, senescence, apoptosis, growth inhibition, etc. The goal of this article is to elucidate whether miR-222 promotes cell proliferation and invasion by targeting RB1, further to explore the molecular mechanism that miR-222 functions as an oncogene in retinoblastoma cells.Methods:miR-222 (miR-222 mimics) and RB1-wt, miR-NC and RB1-wt, miR-222 and RB1-mut, miR-NC (a controlled miR-222 mimics) and RB1-mut were co-transfected into Y79 cells, and luciferase activity was detected by single photon. Retinoblastoma cells were transfected with miR-222 mimics and miR-NC, and the expressions of RB1 protein were detected by Western blot. Retinoblastoma cell proliferation assays were performed by MTS assay when miR-222, miR-NC, RB1 (pcDNA3.1-RB1), vector (pcDNA3.1), miR-222+RB1 and miR-NC+vec-tor were transfected into Y79 cells. The growth and invasion ability of Y79 cells with ectopic expression of miR-222 were evaluated by MTS and Transwell invasion assays.Results:This study demonstrated that miR-222 could promote the luciferase activity of RB1-wt. The expression levels of luciferase reporter gene activity in Y79 cells after transfection with miR-222+RB1-wt were higher than those in the negative control cells (miR-NC+RB1-wt) (P0.05). After transfection with miR-222 mimics for 48 h, Transwell invasion assay showed that the number of cells through the basement membrane was (193±10). Compared with the control group (144±11), it could signiifcantly accelerate the invasion of Y79 cells (P0.05).Conclusion:miR-222 promotes cell proliferation and invasion by targeting RB1 expression in retinoblastoma cells.

Progress of miR-221/222 in tumor diagnosis and treatment / 中国基层医药

MicroRNAs(miRNA)are a kind of small non -conding RNA,which negative regulate target genes expression at post -transcription level by binding 3'-untraslation region(3'UTR).Dysregulation of miRNA pro-files relate to cancer,immune disorders,cardiovascular disease,neurodegenerative diseases and metabolic diseases. miR -221 /222 are two similary miRNAs,which share the same promoter and have the same seed sequence.miR -221 /222 usually target to same target genes and co -regulate same processes and act as onco -miRNA roles.Howev-er,miR -221 /222 maybe act as suppressor in cancer which maybe dependent particular context.The roles of miR -221 /222 are reviewed in this article,will provide a comprehensive vision for comprehending the biological process of miR -221 /222 in carcinoma.

Effect of up-regulating miR-222 in HepG2 cells on its invasion and proliferation / 实用肿瘤学杂志

Objective To detect the changes of cell invasion and proliferation in hepatocellular carcino -ma cell line HepG2 through the upregulation of miR -222.Methods The expression of miR-222 was up-reg-ulated by liposomal transfection .The expression of PTEN which is the targeting protein of miR -222 was detected by using Western Blot assay .The changes of proliferative capacity and invasive ability in HepG 2 cells after the upregulation of miR-222 were detected through MTT assay and Transwell assay .Results In Western Blot ex-periments,the expression of PTEN in transfection groups were lower than that in negative control groups and blank control groups.In MTT assay,the OD values in transfected group were higher than that in negative control group and blank control group(P<0.05).In transwell experiments,the number of penetrating cells in transfected group were higher than that in negative control group and blank control group (P<0.05).Conclusion Our results sug-gest that PTEN is significantly down regulated by the upregulation of miR -222.The proliferative capacity and inva-sive ability are raised in hepatocellular carcinoma cell line HepG 2 through the upregulation of miR-222.

miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4 / 中国癌症杂志

Background and purpose:MicroRNA (miRNA, miR) plays an important regulatory role in cancer. miR-222 is reported to be up-regulated in various tumors, but its role in renal cell carcinoma (RCC) remains unclear. In this study, we detected the expression of miR-222 in both RCC and adjacent tissue samples. The aim of this study was to investigate the role of miR-222 in RCC. Methods:The expression levels of miR-222 in RCC tissue samples were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). DDIT4 and LC3-Ⅱ protein expressions were determined by Western blot. Dual luciferase assay was performed to verify the target of miR-222. EGFP-LC3 microscopy assay was performed to assess autophagy. Results:Results from qRT-PCR showed that the expression of miR-222 was up-regulated in RCC tissues. Knockdown of miR-222 with speciifc antagomiR decreased the cell viability of 786-O cells, whereas overexpression of miR-222 increased the cell viability (P<0.01). The levels of DDIT4 were up-regulated in 786-O cells transfected with miR-222 antagomiR, whereas overexpression of miR-222 induced the down-regulation of DDIT4 expression. Data from dual luciferase assay indicated that miR-222 directly targeted the expression of DDIT4. Consistently, the expression of DDIT4 in RCC tissues was down-regulated compared with adjacent tissues. Knockdown of miR-222 in 786-O cells induced a signiifcant increase of autophagosome formation and LC3 lipidation.These results supported that miR-222 could inhibit autophagy in RCC cells, which may affect the clinical characteristcs of RCC. Conclusion: miR-222 is up-regulated in RCC and can inhibit the autophagy of RCC cells through down-regulating the expression of DDIT4.

Discriminating thyroid cancers from benign lesions based on differential expression of a limited set of miRNA using paraffin embedded tissues.

Background : Micro-RNAs (miRNAs) are expressed in a tissue-specific manner and are known to demonstrate differential expression even among the various subtypes of a given tumor. This differential expression has been harnessed successfully in the development of diagnostic assays for various malignant tumors. These assays have been found to be relevant and of value as additional diagnostic tools even among thyroid tumors, particularly with regard to thyroid carcinomas of follicular morphology. Materials and Methods : A limited set of miRNA have been assessed as part of this study in an effort to use minimal number of miRNA markers (miR-187, miR-221, miR-222, and miR-224) to differentiate the benign from the malignant thyroid tumors using miRNA derived from paraffin embedded material. Results : While miR-221 and miR-222 were found to provide good accuracy as individual markers (86% and 84%), a combination of the two provided slightly better accuracy (91%). Both miR-221 and 222 were able to significantly differentiate malignant tumors from the benign samples (P< 0.001) individually and as a combination of markers. However, inclusion of miR-187 and miR-224 in the panel did not provide any additional benefit. Conclusion : While a combination of miR-221 and 222 when used in a diagnostic panel could provide fairly good accuracy additional markers may need to be investigated to augment their diagnostic utility.