CircRNA-SCAF8 promotes vascular endothelial cell pyroptosis by regulating the miR-93-5p/TXNIP axis / 浙江大学学报·医学版

OBJECTIVES@#To investigate the role and mechanism of circRNA-SR-related CTD associated factor 8 (SCAF8) in regulating endothelial cell pyroptosis in high glucose environment.@*METHODS@#Human umbilical vein endothelial cells (HUVECs) were cultured and divided into six groups. The normal control group and high glucose control group were cultured in cell culture medium with 5 and 33 mmol/L glucose, respectively. The RNA control group, circRNA-SCAF8 inhibition group, miR-93-5p overexpression group and miR-93-5p inhibition group were added with non-functional siRNA, circRNA-SCAF8 inhibitor, miR-93-5p overexpression molecule and miR-93-5p inhibitor in high glucose environment, respectively. Cell viability and pyroptosis were detected by cell counting kit-8 (CCK-8) assay, flow cytometry and Hoechst 33342/propidium iodide fluorescence double staining. Western blotting and enzyme-linked immunosorbent assay were used to detect the expression of pyroptosis-related factors including apoptosis-associated speck-like protein containing a CARD (ASC), cysteine aspartic acid specific protease-1 (caspase-1) and Gasdermin D (GSDMD), NOD like receptor protein 3 (NLRP-3), thioredoxin interacting proteins (TXNIP), IL-18 and IL-1β. The expression of circRNA-SCAF8, miR-93-5p and TXNIP was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Fluorescence in situ hybridization (FISH) was used to locate circRNA-SCAF8 and miR-93-5p. Dual luciferase assay was used to verify the targeted regulatory relationship between miR-93-5p and upstream and downstream molecules.@*RESULTS@#Compared with the RNA control group, the cell survival rate of circRNA-SCAF8 inhibition group and miR-93-5p overexpression group increased (both P<0.01), the pyroptosis decreased (both P<0.01), and the expressions of pyroptosis-related factors such as TXNIP, NLRP-3, caspase-1, GSDMD, ASC, IL-18 and IL-1β were significantly decreased (all P<0.05). The expression of miR-93-5p was significantly increased after inhibition of circRNA-SCAF8 (P<0.01), and the expression of circRNA-SCAF8 tended to decrease after overexpression of miR-93-5p, but with no statistical significance (P>0.05). Dual luciferase assay showed that miR-93-5p downre-gulated circRNA-SCAF8 expression by binding to the 3 ´ UTR region of circRNA-SCAF8, and miR-93-5p downregulated TXNIP expression by binding to the 3 ´ UTR region of TXNIP. FISH showed that circRNA-SCAF8 and miR-93-5p were both located in the cytoplasm and were highly associated in the cells. qRT-PCR showed that the relative expression of TXNIP increased or decreased after overexpression or inhibition of miR-93-5p compared with the RNA control group, respectively (both P<0.05), suggesting that miR-93-5p could regulate TXNIP gene expression.@*CONCLUSIONS@#CircRNA-SCAF8/miR-93-5p/TXNIP axis is involved in the regulation of pyroptosis in HUVECs under high glucose.

Expression significance of serum miR-27, miR-93, BMP-15 in patients with polycystic ovary syndrome / 中华内分泌外科杂志

Objective:To explore the expression significance of serum MicroRNA-27 (miR-27), MicroRNA-93 (miR-93) and bone morphogenetic protein 15 (BMP-15) in patients with polycystic ovary syndrome (PCOS) .Methods:63 PCOS patients admitted to Yantai Yantaishan Hospital from Aug. 2015 to Jan. 2019 were selected and divided into the obese group (obese PCOS, BMI≥25 kg/m 2, n=22) and the non-obese group (non-obese PCOS, BMI<25 kg/m 2, n=41) according to body mass index (BMI). 30 healthy women with normal physical examination during the same period were selected as the healthy group. Serum miR-27, miR-93 expression levels and bone morphogenetic protein 15 (BMP-15) concentration of the three groups were analyzed, and endocrine metabolism indicators of the three groups were compared. Receiver operating characteristic curve (ROS) was used to evaluate the predictive value of serum miR-27, miR-93 and BMP-15 for PCOS and obese PCOS patients. Results:(1) Endocrine indicators: Compared with the healthy group, FSH was lower in the obese group and the non-obese group, and T, LH/FSH and IR were higher ( P<0.05) ; Compared with the non-obese group, T and IR were higher in the obese group ( P<0.05) ; (2) Serum miR-27, miR-93 and BMP-15: Compared with the healthy group, serum BMP-15 concentration was lower in the obese group and the non-obese group, and serum miR-27, miR-93 expression levels were higher ( P<0.05) ; Compared with the non-obese group, the serum miR-27 and miR-93 expression levels in the obese group were higher ( P<0.05) ; (3) miR-27, miR-93 and BMP-15 had predictive value for PCOS, and the area under the curve of BMP-15 was the highest ( P<0.05) ; (4) miR-27 and miR-93 had predictive value for obese PCOS, and the area under the curve of miR-93 was the highest ( P <0.05) . Conclusions:In addition to significant endocrine index disorders in obese PCOS patients, serum miR-27 and miR-93 are highly expressed and the level of BMP-15 is relatively low. BMP-15 can be used as an effective parameter to assist in the diagnosis of PCOS, and miR-93 can be used as an effective parameter to assist in the diagnosis of obese PCOS.

miR-93 activates ERK pathway to promote proliferation and migration of NSCLC cells via targeting EphA4 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of miR-93/EphA4 (Eph receptor A4) axis on the proliferation and migration of nonsmall cell lung cancer (NSCLC) H460 and H1299 cells via regulating extracellular regulated protein kinases (ERK) pathway. Methods: The expression levels of miR-93 in H460 and H1299 cells was detected by qPCR. miR-93 mimics and EphA4 overexpression plasmids were transfected into H460 cells and miR-93 inhibitor was transfected into H1299 cells respectively, after which MTT assay and Transwell assay were used to detect the effects of miR-93 on proliferation and migration of transfected cells. The targeted regulatory relationship betweenmiR-93andEphA4wasverifiedbyDual-luciferasereportergeneassay.Theexpression levels of PCNA(proliferating cell nuclear antigen), EphA4, ERK and p-ERK were detected by Westernblotting.The effects of simultaneous overexpression of miR-93 and EphA4 on proliferation and migration of H460 cells were detected by MTT assay and Transwell assay. Results: The expression of miR-93 in H1299 cells was higher than that in H460 cells (P<0.01). Overexpression of miR-93 promoted proliferation and migration of H460 cells (all P<0.01), and knockdown of miR-93 inhibited proliferation and migration of H1299 cells (all P<0.01). The Dualluciferase reporter gene assay confirmed that miR-93 could target EphA4. Overexpression of miR-93 down-regulated the mRNA and protein expression levels of EphA4(allP<0.05), and promoted proliferation and migration of H460 cells through targeted regulation of EphA4 and activation of ERK pathway (all P<0.01). Conclusion: miR-93 promotes the proliferation and migration of NSCLC cells, and its mechanism may be related to the targeted regulation of EphA4 and activation of the ERK pathway.

Prediction of retinopathy in type 2 diabetes mellitus by combined detection of microRNA93 and microRNA21 / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the predictive value of combined detection of plasma microRNA-93(miR-93)and microRNA-21(miR-21)in the progression of type 2 diabetic retinopathy(DR).<p>METHODS: Totally 76 patients with type 2 diabetic retinopathy admitted to our hospital from June 2013 to June 2014 were divided into DR non-progressive group(34 cases)and DR progressive group(42 cases)according to the follow-up results, and 45 healthy people were selected as the control group. The serum levels of miR93 and miR21 in three groups were detected; the independent risk factors of prognosis in DR patients were analyzed; and the predictive value of miR93 and miR21 in DR patients was analyzed.<p>RESULTS: The serum levels of miR93 and miR21 in DR progressive group were significantly higher than those in non-progressive group and control group(all <i>P</i>< 0.01); both of them could be used as independent risk factors affecting the progress of DR patients; the area, specificity and sensitivity under the combined detection curve were higher than those of single detection(<i>P</i><0.05).<p>CONCLUSION: The serum levels of miR93 and miR21 are increased in DR patients, which can affect the progress of DR patients, and can be used as biomarkers for the diagnosis of DR.

Association of insulin sensitivity and β-cell function with body fat in patients with PCOS / 中华内分泌代谢杂志

A total of 187 patients with polycystic ovary syndrome (PCOS) in our hospital from January 2016 to December 2018 were enrolled, and 190 healthy people served as control group. The levels of homeostasis model assessment of insulin resistance index ( HOMA-IR), β-cell function index ( HOMA-β), total cholesterol ( TC), triglyceride (TG), low density lipoprotein-cholesterol ( LDL-C), high density lipoprotein-cholesterol ( HDL-C), body fat content ( BF), and miR-93 were compared between the two groups. The results showed that HOMA-IR, HOMA-β, TG, TC, LDL-C, BF, and miR-93 in PCOS group were significantly higher while HDL-C was significantly lower than those in control group (all P＜0.05). HOMA-IR, HOMA-β, TG, TC, and LDL-C levels in patients with Fat≥35% of PCOS group were significantly higher compared with those in patients with BF＜35% ( P＜0.05) while HDL-C was significantly lower (P＜0.05). There were no significant differences in TC and miR-93 between patients with BF≥35% and Fat＜35% in PCOS group (P＞0.05). HOMA-IR and HOMA-β were positively correlated with BF level (r=0.427 and 0.224, P＜0.05), while miR-93 was not correlated with BF level (P＞0.05).

miR-93-5p Transferred by Exosomes Promotes the Proliferation of Esophageal Cancer Cells via Intercellular Communication by Targeting PTEN / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the relationship between plasma miR-93-5p and the risk of esophageal cancer, as well as the influence of miR-93-5p on the biological function of esophageal cancer cells, exerted through exosomes.</p><p><b>METHODS</b>The expression of plasma miR-93-5p in esophageal cancer patients and healthy controls was analysed by real-time quantitative PCR. The influence of miR-93-5p on the risk and prognosis of esophageal carcinoma was analyzed by conditional logistic regression and survival analysis. The effect of miR-93-5p on the biological function of recipient cells was investigated by establishing an in vitro donor cell co-culture model. The target gene of miR-93-5p was validated by luciferase reporter assay and Western Blotting.</p><p><b>RESULTS</b>Upregulation of plasma miR-93-5p expression significantly increases the risk of esophageal cancer and is associated with poor prognosis. miR-93-5p transferred by exosomes promotes the proliferation of recipient esophageal cancer cells and affects the expression of PTEN and its downstream proteins p21 and cyclin D1.</p><p><b>CONCLUSION</b>Our study provides a reference for the identification of biomarkers for the diagnosis and prognosis of esophageal cancer.</p>

Expression of miR-93 and miR-21 in patients with diabetic retinopathy and its clinical values / 眼科新进展

Objective To investigate the expression and clinical value of plasma miR-93 and miR-21 in patients with diabetic retinopathy (DR).Methods Ninety-eight patients with DR (DR group),40 patients with type 2 diabetes mellitus (DM group) and 40 healthy subjects (control group) from January 2015 to March 2017 were enrolled in this study.And the expression levels of miR-93 and miR-21 in the plasma were detected by real-time quantitative PCR (RT-qPCR) techniques.ROC curve was used to evaluate the values of miR-93 and miR-21 for the diagnosis of DR,and the risk factors of DR were analyzed by multivariate logistic regression analysis.Finally,Pearson correlation analysis was utilized to evaluate the correlation of plasma miR-93 and miR-21 with biochemical indicators including total cholesterol (TC),low density lipoprotein cholesterol (LDL-C),triglyceride (TG) and creatinine (Cr),blood urea nitrogen (BUN),urinary albumin/creatinine (UACR),glycosylated hemoglobin (HbAlc) and fasting blood glucose (FPG) in DR patients.Results Both DR and DM groups had significantly higher UACR,HbAlc and FPG than the control group (all P ＜ 0.05),and the DR group was significantly higher than DM group (all P ＜ 0.05).The expression levels of plasma miR-93 and miR-21 in the DR group (1.84 ±0.27,1.95 ±0.34) were significantly higher than those in the DM group (1.09 ± 0.13,1.13 ± 0.18) and the control group (1.02 ± 0.08,1.08 ±0.16),and there were statistical differences (all P ＜ 0.05).Meanwhile,the levels of plasma miR-93 and miR-21 in PDR group (2.31 ± 0.57,2.28 ± 0.61) were significantly higher than those in NPDR group (1.40 ± 0.21,1.52 ± 0.24),and there were statistical differences (all P ＜ 0.05).The area under the curve (95％ CI) of plasma miR-93,miR-21 and both indicators for DR diagnosis was 0.892 (0.824-0.961),0.836 (0.770-0.905),0.937 (0.868-0.992),respectively.Multivariate logistic regression analysis showed that UACR,HbAlc,FPG,miR-93 and miR-21 were independent risk factors for DR.Correlation analysis showed that plasma miR-93 and miR-21 were positively correlated with UACR,HbAlc and FPG in the patients with DR (all P ＜ 0.01),and miR-93 was positively correlated with miR-21 (P ＜ 0.01).Conclusion Plasma miR93 and miR-21 are closely related to the initiation and development of DR and can be used as the potential markers for the diagnosis of DR.

Expression of miR-93-5p and miR-27a-3p in rectal cancer tissues and its clinical significance / 中华放射肿瘤学杂志

Objective To investigate the miRNA expression profiles in rectal cancer tissues and their associations with clinical pathological stage, depth of tumor invasion, and lymph node metastasis, and to evaluate the potential of miRNA as diagnostic and prognostic markers of rectal cancer. Methods Human miRNA microarray was used to profile miRNA expression in rectal cancer tissues and matched adjacent normal tissues (n=71). The up-regulated miR-93-5p and down-regulated miR-27a-3p were screened out, and the top differentially expressed miRNA were validated by quantitative real-time polymerase chain reaction ( qRT-PCR) . The relationship between the expression of miRNA and clinical parameters was analyzed by ANOVA and Spearman correlation. Results The expression of miR-27a-3p was down-regulated in miRNA microarray, but was up-regulated in qRT-PCR analysis;the data were relatively discrete. The expression of miR-93-5p was up-regulated in both miRNA microarray and qRT-PCR analysis;the expression level of miR-93-5p in rectal cancer tissues was 3165 times that in adjacent normal tissues ( P=00058);the expression level was correlated with tumor volume ( P= 0004 ) , and was positively correlated with the level of carcinoembryonic antigen ( CEA) before treatment ( P=0001) and the number of lymph nodes metastases (rs=0534, P=0005). Conclusions There is a differential miRNA expression pattern between rectal cancer tissues and matched adjacent normal tissues. The miR-93-5p is highly up-regulated in rectal cancer tissues and may serve as a diagnostic and prognostic marker of rectal cancer.

Expression of miR-93-5p and miR-27a-3p in rectal cancer tissues and its clinical significance / 中华放射肿瘤学杂志

Objective To investigate the miRNA expression profiles in rectal cancer tissues and their associations with clinical pathological stage, depth of tumor invasion, and lymph node metastasis, and to evaluate the potential of miRNA as diagnostic and prognostic markers of rectal cancer. Methods Human miRNA microarray was used to profile miRNA expression in rectal cancer tissues and matched adjacent normal tissues (n=71). The up-regulated miR-93-5p and down-regulated miR-27a-3p were screened out, and the top differentially expressed miRNA were validated by quantitative real-time polymerase chain reaction ( qRT-PCR) . The relationship between the expression of miRNA and clinical parameters was analyzed by ANOVA and Spearman correlation. Results The expression of miR-27a-3p was down-regulated in miRNA microarray, but was up-regulated in qRT-PCR analysis;the data were relatively discrete. The expression of miR-93-5p was up-regulated in both miRNA microarray and qRT-PCR analysis;the expression level of miR-93-5p in rectal cancer tissues was 3165 times that in adjacent normal tissues ( P=00058);the expression level was correlated with tumor volume ( P= 0004 ) , and was positively correlated with the level of carcinoembryonic antigen ( CEA) before treatment ( P=0001) and the number of lymph nodes metastases (rs=0534, P=0005). Conclusions There is a differential miRNA expression pattern between rectal cancer tissues and matched adjacent normal tissues. The miR-93-5p is highly up-regulated in rectal cancer tissues and may serve as a diagnostic and prognostic marker of rectal cancer.

Target genes of the hsa-miR-106b-25 cluster:Prediction and bioinformatic analysis / 医学研究生学报

Objective The hsa-miR-106b-25 gene cluster is involved in various biological processes of carcinoma .This study aims at a prediction and function analysis of the hsa-miR-106b-25 gene cluster, miR-106b, miR-93, and miR-25, so as to provide some evidence for further studies on the functions of the three miRNAs and the mechanisms of their interaction . Methods We obtained the sequences of miR-106b, miR-93, and miR-25 from the miRBase, predicted their target genes with TargetScan , PicTar, and miRanda, and used 3 or more experimentally verified target genes from the miRTarbase as the gene set for further bioinformatic analysis .We predic-ted the biological processes of the target genes by GeneOntology analysis and enriched KEGG ( Kyoto Encyclopedia of Genes and Genomes) by pathway analysis, produced protein-protein interaction ( PPI) networks with STRING . Results The target genes of the miR-106b-25 gene cluster were significantly enriched in such biological processes as the regulation of macromolecule metabolism , regulation of metabolic process , and cell cycle process , while the KEGG pathway mainly in glioma, melanoma, prostate cancer , and gallbladder carcino-ma.The proteins encoded by the targeted genes of showed complicated interactions , and those encoded by the KAT2B, PTEN, TP53, CDH1, MDM2, E2F1, RB1, and SMAD7 plaid a core role in the interac-tion network. Conc lusoi n MiRNAs of the miR-106b-25 gene cluster regulate the downstream target proteins involved in tumorigenesis by participating in the cell cycle and cancer signaling pathway .

Function and Regulation of miR-9-3 in Chronic Lymphocytic Leukemia / 现代检验医学杂志

Objective To investigate the role and regulatory mechanism of micro RNA-9-3 (miR-9-3)in the pathogenesis of chronic lymphocytic leukemia.Methods Using the methylation specific PCR (MSP)technology to detect 8 cases of normal bone marrow tissue and peripheral blood,78 cases of bone marrow tissue came from the chronic lymphocytic leukemia pateints newly diagnosed and the methylation level of 7 kinds of leukemia cell line.Used Western blot to detected the NF-kappa B1 signal transduction pathway activation levels of methylation positive leukemia cell line.Results The miR-9-3 of normal control group were in the negative methylation status.Only I83-E95 and WAC3CD5+ were in positive methylation status in seven kinds of leukemia cell line (the positive of MSP was 28.6%);65 cases occurred miR-9-3 methylated in 78 of chronic lymphocytic leukemia patients (the positive of MSP was 83%).I83-E95 and miR-9-3 cells of WAC3CD5+ were in the methylation state when treatment with 5-nitrogen-2’-deoxidization cytidine (5-Aza2’Dc).Conclusion The abnormal methylation of miR-9-3 were usually seenin chronic lymphocytic leukemia,it could lead to abnormal hyperplasis in cancer cells.The methylation of miR-9-3 could inhibit the activation of NF-kappa B1 signal pathway suggested that it could sup-press the apoptosis of cancer cells through this pathways to trogered the progression of disease.The inhibitor of methylation could be induced the demethylation of leukemia cell lines,so it is possible that miR-9-3 maight be a new gene targets for the treatment of chronic lymphocytic leukemia.