Expression and clinical significance of serum miR-221 and miR-127 levels in children with severe pneumonia / 中国医师杂志

Objective:To analyze the expression and clinical significance of serum microRNA-221 (miR-221) and microRNA-127 (miR-127) levels in children with severe pneumonia.Methods:151 children with severe pneumonia treated in Handan Central Hospital from January 2017 to December 2019 were prospective selected as severe pneumonia group. According to the prognosis of children in hospital, they were divided into death group (16 cases) and survival group (135 cases). Another 80 healthy children who underwent physical examination in our hospital during the same period were selected as normal control group. Microarrays were used to analyze the expression of miRNAs in the severe pneumonia group and the normal control group, and the miRNAs with the most significant up-regulation or down-regulation (based on the difference expression change ≥2 times threshold and P<0.05) were selected. The serum miR-221, miR-127 and inflammatory factor [C-reactive protein (CRP), calcitonin (PCT) and interleukin 6 (IL-6), interleukin 8 (IL-8), tumor necrosis factor-α (TNF-α)] levels were detected in the enrolled subjects. The correlation between serum miR-221 and miR-127 and the above inflammatory indicators in severe pneumonia group were analyzed. The diagnostic value and prognostic value of miR-221 and miR-127 in severe pneumonia were analyzed by receiver operating characteristic (ROC) curve. Results:The serum levels of miR-221 and miR-127 in severe pneumonia group were lower than those in normal control group, while the serum levels of CRP, PCT, IL-6, IL-8 and TNF-α were higher than those in the normal control group (all P<0.05). Serum miR-221 was negatively correlated with CRP, PCT, TNF-α (all P<0.05); Serum miR-127 was negatively correlated with CRP, PCT, IL-6 and IL-8 (all P<0.05). The serum levels of miR-221 and miR-127 in the death group were lower than those in the survival group ( P<0.05). The area under the curve (AUC) of miR-221 and miR-127 for the combined diagnosis of severe pneumonia was 0.871, and the AUC for the prognosis of severe pneumonia was 0.851. Conclusions:The serum levels of miR-221 and miR-127 in children with severe pneumonia decreased, which was negatively correlated with the levels of some inflammatory factors. The combined detection of miR-221 and miR-127 has high clinical value in the diagnosis and prognosis prediction of severe pneumonia.

Preparation of folate-targeted anti-miR- 221 anionic liposome and its in vitro anti-hepatocellular carcinoma effect / 中国药房

OBJECTIVE To prep are folate-targeted miR- 221 antisense oligonucleotide （anti-miR-221）delivery system ，and to preliminarily evaluate its in vitro anti-cancer effect on hepatocellular carcinoma. METHODS Folate-targeted anti-miR- 221 liposomes（FRL）were prepared by thin-film dispersion method ；the particle size ，Zeta potential and encapsulation efficiency were determined. The delivery efficiency of folate-targeted anionic liposome in human hepatoma HepG 2 cells was determined by in vitro cellular uptake experiment using calcein as the model drug. Flow cytometry was used to detect the effects of FRL on the apoptosis and cell cycle of HepG 2 cells. RESULTS The particle size of prepared FRL was （172.70±3.76）nm，Zeta potential was （－1.16± 0.15）mV and encapsulation efficiency was （83.53±1.85）％. In vitro cellular uptake experiments showed that folate-targeted anionic liposome successfully delivered calcein to HepG 2 cells，and the delivery efficiency in targeted group was higher than that of non-targeted liposome group （P＜0.01）. Apoptosis experiment results showed that the apoptotic rate of HepG 2 cells treated with FRL was significantly higher than that of non-targeted liposome （P＜0.01）. In cell cycle experiment ，FRL could shorten the S phase fraction of HepG 2 cells and induced arrest in the G 0/G1 and G 2/M phases. CONCLUSIONS FRL can encapsulate anti-miR-221 well and deliver it to hepatocellular carcinoma HepG 2 cells successfully ，and has a good in vitro anti-hepatoma effect in inducing apoptosis and cell cycle regulation.

Exosomes Derived from Hydroquinone-transformed Human Bronchial Epithelial Cells Inhibited Recipient Cell Apoptosis by transferring miR-221 / 生物医学与环境科学（英文）

Objective@#Although benzene is a confirmed environmental carcinogen, the mechanism of its carcinogenicity remains largely unclear. The suggested oncogene, miR-221, is elevated and plays important roles in various tumors, but its role in benzene-induced carcinogenesis remains unknown.@*Methods@#In the present study, we constructed hydroquinone (HQ, a representative metabolite of benzene with biological activity)-transformed malignant cell line (16HBE-t) and analyzed the level of miR-221 in it with qRT-PCR. Exosomes from 16HBE-t cells incubated with or without an miR-221 inhibitor were isolated by ultracentrifugation, characterized by transmission electron microscopy and laser scanning confocal microscope, and then transfected into 16HBE cells. The effects of exosomal miR-221 on apoptosis induced by HQ in recipient cells were determined using flow cytometry.@*Results@#The amount of miR-221 in 16HBE-t was significantly increased compared with controls. When recipient cells ingested exosomes derived from 16HBE-t, miR-221 was increased, and apoptosis induced by HQ was inhibited. Blocking miR-221 in 16HBE-t using an inhibitor did not significantly alter miR-221 or apoptosis in recipient cells.@*Conclusion@#Exosomal miR-221 secreted by 16HBE-t inhibits apoptosis induced by HQ in normal recipient cells.

Anti-micro rna-221 a promising genetic therapy of oral squamous cell carcinoma (scc-25)

Abstract Micro-RNA-221(miR-221) is one of oncogenic miRNAs that plays a vital role in the development and progression of oral cancers. The aim of this study is to introduce a new gene therapy for oral squamous cell carcinoma by blocking the expression of oncogenic miR-221 by its inhibitor. The present work was performed on squamous cell carcinoma cell line SCC-25 and anti-miR-221 was delivered to the cells using an ultrasound micro bubbles. Assessment of the effect of miR-221 inhibitor on SCC-25 cells was done using MTT assay, cell cycle analysis and apoptosis detection. In addition, reverse transcription-polymerase chain reaction was also used to detect the expression -miR-221 and its target genes. Using ANOVA, statistical analysis of the results showed significant inhibition of cell viability with and induction of cell apoptosis of SCC-25 cell line after transfection. Moreover, the expression of miR-221, Epidermal growth factor receptor (EGFR) and CDKNIB/p27 were downregulated without significant difference. Transfection of SCC-25 by inhibitor of miR-221 resulting in blockage of its expression leading to arresting of tumor growth. These results proved the effective role of micro-RNA inhibitors as novel therapeutic agent for oral cancers.

Resumo Micro-RNA-221 (miR-221) é um dos miRNAs oncogênicos que desempenham um papel vital no desenvolvimento e progressão de carcinomas orais. O objetivo deste estudo é apresentar uma nova terapia gênica para o carcinoma epidermóide oral por meio do bloqueio da expressão do miR-221 oncogênico por seu inibidor. O presente trabalho foi realizado na linhagem de células de carcinoma de células escamosas SCC-25 e o anti-miR-221 foi administrado às células usando micro-bolhas de ultrassom. A avaliação do efeito do inibidor miR-221 em células SCC-25 foi feita usando ensaio de MTT, análise do ciclo celular e detecção de apoptose. Além disso, a reação em cadeia da polimerase com transcrição reversa também foi usada para detectar a expressão -miR-221 e seus genes-alvo. Usando ANOVA, a análise estatística dos resultados mostrou inibição significativa da viabilidade celular e indução da apoptose celular da linhagem celular SCC-25 após a transfecção. Além disso, a expressão de miR-221, receptor do fator de crescimento epidérmico (EGFR) e CDKNIB/p27 foram regulados para baixo sem diferença significativa. A transfecção de SCC-25 por inibidor de miR-221 resultou no bloqueio de sua expressão, levando à interrupção do crescimento do tumor. Esses resultados comprovaram o papel eficaz dos inibidores de micro-RNA como novo agente terapêutico para carcinomas orais.

Molecular mechanism of miR-221 promoting apoptosis of human retinal vascular endothelial cells induced by high glucose by regulating p53/MDM2 signaling pathway / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the effect of miR-221 on apoptosis of high glucose-induced human retinal vascular endothelial cells and to explore its mechanism. <p>METHODS: High-glucose-induced HRCECs were established by treatment of HRCECs cells with glucose at 30mmol/L for 48h; HG+miR-NC group(transfected miR-NC), HG+miR-221 group(transfected miR-221 mimics), HG+anti-miR-NC group(transfected anti-miR-NC), HG+anti-miR-221 group(transfected anti-miR-221), HG+miR-221+pcDNA 3.1 group(co-transfected miR-221 mimics and pcDNA 3.1), HG+miR-221+pcDNA 3.1-MDM2 group(co-transfected miR-221 mimics and pcDNA 3.1-MDM2), transfected into HRCECs cells by liposome method, and then treated with high glucose; qRT-PCR method for detection the expression of miR-221, p53 and MDM2; the protein expression of p53 and MDM2 were detected by Western blot. The apoptosis of cells was detected by flow cytometry. <p>RESULTS: Compared with NG group, the expression of miR-221 and p53 was significantly increased, the expression of MDM2 was significantly decreased, and the apoptosis rate was significantly increased in high glucose-induced HRCECs. Overexpression of miR-221 induced apoptosis of high glucose-induced HRCECs cells is more obvious. Inhibition of miR-221 can down-regulate the apoptosis of high glucose-induced HRCECs and down-regulate p53, up-regulate MDM2; overexpression of MDM2 can reverse the inhibition by miR-221 anti-apoptotic effect of cells and regulation of p53 and MDM2 of high-glucose-induced HRCECs. <p>CONCLUSION: miR-221 can promote the apoptosis of high-glucose-induced human retinal vascular endothelial cells, and its mechanism is related to p53/MDM2 signaling pathway.

Serum levels and clinical significances of miR-205 and miR-221 in patients with colon cancer / 国际肿瘤学杂志

Objective@#To investigate the serum levels and clinical significances of microRNA-205 (miR-205) and microRNA-221 (miR-221) in patients with colon cancer.@*Methods@#A total of 172 patients with colon cancer (colon cancer group), 130 patients with benign diseases of colon (benign lesion group) and 70 healthy persons (control group) admitted to Central Hospital of Western Hainan from January 2016 to December 2018 were selected. The serum levels of miR-205 and miR-221 in each group were detected, and their relationships with clinicopathological characteristics of patients with colon cancer were analyzed. The diagnostic values of the serum levels of miR-205 and miR-221 in colon cancer were analyzed by receiver operating characteristic (ROC) curve. Pearson correlation analysis was used to analyze the correlation between the serum levels of miR-205 and miR-221.@*Results@#The serum levels of miR-205 in colon cancer group, benign lesion group and control group were 2.84±0.96, 1.16±0.27 and 1.05±0.23, with a statistically significant diffe-rence (F=10.113, P<0.001). The serum levels of miR-221 in the three groups were 1.95±0.74, 0.37±0.08 and 0.32±0.05, with a statistically significant difference (F=12.416, P<0.001). Further pairwise comparisons found that the serum levels of miR-205 and miR-221 in colon cancer group were significantly higher than those in benign lesion group and control group (all P<0.001). The serum levels of miR-205 and miR-221 in patients with colon cancer were related with TNM stage (t=5.412, P<0.001; t=6.103, P<0.001), degree of tumor differentiation (t=4.573, P=0.028; t=4.805, P=0.013) and with or without lymph node metastasis (t=5.837, P<0.001; t=7.410, P<0.001). ROC curve analysis showed that the optimal cut-off values of the serum levels of miR-205 and miR-221 for the diagnosis of colon cancer were 2.17 and 1.30, respectively. The area under the curve of the two combined diagnostic method for colon cancer was the largest (0.924, 95%CI: 0.865-0.983), with the sensitivity and specificity of 91.3% and 87.4%. The correlation analysis showed that the serum levels of miR-205 and miR-221 were positively correlated in patients with colon cancer (r=0.837, P<0.001).@*Conclusion@#The serum levels of miR-205 and miR-221 are significantly increased in patients with colon cancer, and the two combined detection has a high value for the diagnosis of colon cancer.

Effects of miR-221 on cell proliferation and apoptosis of chronic myeloid leukemia K562 cells and its mechanism / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of down-regulation of miR-221 on cell proliferation and apoptosis of chronic myeloid leukemia (CML) K562 cells and its related regulatory mechanism. Methods: K562 cells were divided into control group, miRNAnegative control (miR-NC) group, miR-221 inhibitor group, miR-221 inhibitor+ negative control siRNA(NC siRNA) group and miR-221 inhibitor+SOCS3 siRNA group. The cells in the control group received no additional treatment. Cells in miR-NC group and miR-221 inhibitor group were transfected with miR-NC and miR-221 inhibitor, respectively. Cells in miR-221 inhibitor+NC siRNA group and miR-221 inhibitor+SOCS3 siRNA group were transfected with NC siRNA and SOCS3 siRNA, respectively, on the basis of successful transfection with miR-221 inhibitor. The transfection efficiency of miR-221 inhibitor was identified by qPCR. Cell viability in each group was measured by CCK-8 assay. Apoptosis in each group was detected by Annexin V-FITC/PI staining using a flow cytometry. The protein expressions of SOCS3, p-JAK1, p-JAK2, p-STAT3 and survivin in each group were detected by WB. Results: Compared with the control group, miR-221 expression was significantly down-regulated in miR-221 inhibitor group (P<0.01), cell viability was significantly reduced at 48 and 72 h after transfection (P<0.05 or P<0.01), the number of apoptotic cells was significantly increased (P< 0.01), the expression of SOCS3 was significantly increased (P<0.01) and the expression levels of p-JAK1, p-JAK2, p-STAT3 and survivin were significantly reduced (all P<0.01). Compared with miR-221 inhibitor group, cell viability was significantly increased at 24, 48 and 72 h after transfection (P<0.05 or P<0.01), the number of apoptotic cells was significantly decreased (P<0.01) and the expression levels of p-JAK1, p-JAK2, p-STAT3 and survivin were significantly increased in miR-221 inhibitor+SOCS3 siRNA group (all P< 0.01). Conclusion: Down-regulation of miR-221 inhibits proliferation and promotes apoptosis of K562 cells, the mechanism of which may be related with up-regulating SOCS3 expression to suppress JAK-STAT3 signaling pathway.

Serum levels and clinical significances of miR-205 and miR-221 in patients with colon cancer / 国际肿瘤学杂志

Objective To investigate the serum levels and clinical significances of microRNA-205 (miR-205) and microRNA-221 (miR-221) in patients with colon cancer.Methods A total of 172 patients with colon cancer (colon cancer group),130 patients with benign diseases of colon (benign lesion group) and 70 healthy persons (control group) admitted to Central Hospital of Western Hainan from January 2016 to December 2018 were selected.The serum levels of miR-205 and miR-221 in each group were detected,and their relationships with clinicopathological characteristics of patients with colon cancer were analyzed.The diagnostic values of the serum levels of miR-205 and miR-221 in colon cancer were analyzed by receiver operating characteristic (ROC) curve.Pearson correlation analysis was used to analyze the correlation between the serum levels of miR-205 and miR-221.Results The serum levels of miR-205 in colon cancer group,benign lesion group and control group were 2.84 ± 0.96,1.16 ± 0.27 and 1.05 ± 0.23,with a statistically significant difference (F =10.113,P ＜ 0.001).The serum levels of miR-221 in the three groups were 1.95 ± 0.74,0.37 ± 0.08 and 0.32 ± 0.05,with a statistically significant difference (F =12.416,P ＜ 0.001).Further pairwise comparisons found that the serum levels of miR-205 and miR-221 in colon cancer group were significantly higher than those in benign lesion group and control group (all P ＜ 0.001).The serum levels of miR-205 and miR-221 in patients with colon cancer were related with TNM stage (t =5.412,P ＜0.001;t =6.103,P ＜0.001),degree of tumor differentiation (t =4.573,P =0.028;t =4.805,P =0.013) and with or without lymph node metastasis (t =5.837,P ＜ 0.001;t =7.410,P ＜ 0.001).ROC curve analysis showed that the optimal cut-off values of the serum levels of miR-205 and miR-221 for the diagnosis of colon cancer were 2.17 and 1.30,respectively.The area under the curve of the two combined diagnostic method for colon cancer was the largest (0.924,95％ CI:0.865-0.983),with the sensitivity and specificity of 91.3％ and 87.4％.The correlation analysis showed that the serum levels of miR-205 and miR-221 were positively correlated in patients with colon cancer (r =0.837,P ＜0.001).Conclusion The serum levels of miR-205 and miR-221 are significantly increased in patients with colon cancer,and the two combined detection has a high value for the diagnosis of colon cancer.

Genistein Improves the Major Depression through Suppressing the Expression of miR-221/222 by Targeting Connexin 43

OBJECTIVE: Recent studies have indicated the possibility that genistein may improve depression via regulating the expression of miR-221/222. This study is to explore whether genistein could improve depression by altering miR-221/222 levels and investigate the possible mechanisms involved in the improvement effect of genistein. METHODS: The animal model of depression was established through unpredictable chronic mild stress. Nest building test and splash test were adapted to evaluate the effects of genistein on depressive symptoms in mice. qRT-PCR and western blot analysis were used to detect the expression of miR-221/222 and connexin 43 (Cx43) in the prefrontal cortex of the mice. In vitro, U87-MG astrocytes were treated with genistein and the expression of miR-221/222 and Cx43 was measured. The dual-luciferase reporter assay was used to verify whether Cx43 was a direct target of miR-221/222. RESULTS: The behavioral tests showed that genistein could significantly reduce depression symptoms of mice, and this remission was not affected by gender. Genistein in vivo and in vitro could reduce increased levels of miR-221 and miR-222 in the prefrontal cortex of depressed mice, while upregulate Cx43 expression. Dual-luciferase reporter assay suggested Cx43 was directly regulated by miR-221/222 in astrocytes. CONCLUSION: Genistein can play its antidepressant effect through down-regulating miR-221/222 by targeting Cx43.

MicroRNA-221/222 participates in the pathogenesis of intrahepatic cholestasis of pregnancy via promoting the apoptosis of human placental trophoblast HTR-8 cells / 中华肝脏病杂志

Objective@#MicroRNA-221/222 is involved in the pathogenesis of intrahepatic cholestasis of pregnancy (ICP) to promote the apoptosis of placental bile acids through human trophoblastic cells. This study investigates the effects of miR-221/222 on proliferation, apoptosis and apoptosis-related proteins of human trophoblast HTR-8/SVneo (HTR-8 cells) to understand its role in promoting trophoblastic apoptosis.@*Methods@#The experiment was divided into transfection group and negative control group. Transient transfection method was used in both groups. The transfection efficiency was detected by RT-QPCR after 48 h transfection. CCK-8 was used to detect the proliferation of HTR-8 cells and the apoptosis of HTR-8 cells were analyzed by flow cytometry. Western blot was used to detect the expression of B-cell Lymphoma 2 (Bcl-2) in HTR-8 cells. Data were compared with t-test.@*Results@#The expression of miR-221/222 transfected group (25.43±0.80, 22.70±0.95) was increased significantly in the HTR-8 cells than that to negative control group (1.14±0.14, 1.58±0.14), and P value was < 0.01, the difference was statistically significant. The expression of Bcl-2 protein in mir-221/222 transfection group was (0.56 ± 0.03, 0.53 ± 0.03), and the protein expression was decreased compared with negative control group (0.72 ± 0.003, 0.76 ± 0.04). P value was < 0.05, the difference was statistically significant, and compared with the mir-221/222 negative control group (8.827 ± 0.48, 11.80 ± 0.45), cell apoptosis of mir-221/222 transfection group (42.53 ± 4.47, 24.09 ± 2.53) increased significantly, P value was < 0.01, and the difference was statistically significant. Proliferation rate in mir-221/222 transfection group was (0.82 ± 0.02, 0.74±0.01), and proliferation was inhibited, when compared with control group (1.15 ± 0.08, 1.06 ± 0.08), P value was < 0.05, and the difference was statistically significant.@*Conclusion@#miR-221/222 may promote the apoptosis of human trophoblastic cells by down regulating the expression of apoptosis inhibitory protein bcl-2, leading to placental dysfunction and impairing the normal bile acid transport function of placenta. This mechanism may be involved in the occurrence and development of ICP.

Effect of procyanidine on the function and expression of miR-221 in human umbilical vein endothelial cells / 解放军医学杂志

Objective To study the effect ofprocyanidine (PC) on the proliferation and migration of human umbilical vein endothelial cells (HUVECs),determine the expression changes of miR-221,and to investigate the mechanism involved.Methods HUVECs were cultured in vitro and treated with PC (5,25,50,75,100μg/ml) for 24 hours,and a PC concentration of 50μg/ml was screened by CCK-8 assay for the follow up experiment,then the cell proliferation activity was detected by the wound healing assay.The expression of miR-221 in HUVECs was detected by real-time quantitative PCR;MiR-221 target genes were predicted in miRWalk database,and the target genes were analyzed by GO and KEGG pathway.Results Compared with the control group,the HUVECs treated with PC for 24h,their proliferation activity in PC 5μg/ml group did not change obviously (P＞0.05),and in PC 25,50,75 and 100μg/ml groups decreased in a concentration dependent manner (P＜0.01).Compared with the control group,the migration ability of PC 50μg/ml group decreased markedly (P＜0.01).Compared with the control group,the expression of miR-221 increased after treatment with PC 50μg/ml (P＜0.01).Go analysis indicated that the target genes of miR-221 were mainly related to cell proliferation,migration,gene translation and so on.The target genes related to cell proliferation and migration were ADAM17,KIT,PDGFD and so on.KEGG pathway analysis showed that the target genes of miR-221 enriched 5 signal pathways,such as FoxO,PI3K-Akt and so on.Conclusions Low concentration of PC has no effect on the proliferation activity of HUVECs.A certain concentration of PC can inhibit the proliferation and migration of HUVECs,of which the mechanism may be involved with the up-regulation of miR-221 and FoxO,PI3K-Akt and other signaling pathways.

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.

Effect of ultrasound microbubble mediated miRNA delivery on the ability of migration ,invasion and clon-ing of human hepatoma HepG2 cells / 实用医学杂志

Objective To invstigate the effect of ultrasound microbubble mediated miRNA delivery on mi-gration,invasion and cloning ability of human hepatoma HepG2 cells. Methods The migration,invasion and col-ony forming ability of HepG2 cells were measured after transfection with antisense miRNA-21/221 and miRNA-199a mimic via the optimal ultrasound microbubble transfection method. Results Compared with the control group ,the migration ,invasion and cloning ability of cells were significantly inhibited after transfection with miRNA mimics(P < 0.05,respectively),especially for miR-199a(relative cell migration rate was 31.05%,the number of invasive cells were 38.67 ± 4.51 and the number of clones were 105.67 ± 5.86). Conclusion The pres-ent study may provide new ideas and clues for gene therapy and prognosis of hepatocell ular carcinoma through ana-lyzing the effect of miRNAs on the biological characteristics of human hepatoma HepG2 cells.

The role of miR-221 in the injury induced by hydrogen peroxide in rat myocardial cells / 中国比较医学杂志

Objective To explore the role of miR-221 in the injury induced by hydrogen peroxide (H2O2) in rat myocardial cells (H9c2).Methods The viability of H9c2 cell induced by cell different concentrations of H2O2 was determined by MTT.The expression of miR-221 was detected by RT-PCR method.The miR-221 inhibitor and negative control were transferred into H9c2 cells by Lipofectamine 2000, then the cells were divided into normal control group, model control group (H2O2 group), negative control group (H2O2+ negative control group), inhibition group (H2O2+miR-221 inhibitor group).The cell viability was measured by MTT assay.Cell apoptosis was detected by acridine orange staining method.The expression of Bcl-2, Bax, phosphatase and tensin homolog deleted on chromosome ten (PTEN, p-protein kinase B (AKT) were assayed by Western Blot.Results 0,25,50,100,200,400 μmol/L H2O2 inhibited H9c2 cell activity gradually, of which 200 mol/L inhibition of cell viability moderate, so as a subsequent induction dose.Compared with normal control group, cell viability was decreased (P < 0.01), cell apoptotic rat was increased (P < 0.01), the expression of Bax and PTEN was upregulated (P < 0.01), the expression of Bcl-2 and p-AKT was downregulated (P < 0.01) in model control group and negative control group.Compared with model control group and negative control group, inhibition group proves the contrary.Conclusions Down-expression of miR-221 could significantly inhibit oxidative stress damage in H9c2 cells, which related to regulation of PTEN/AKT signal pathway.

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.

Effects of α-pinene extracted from pine needle on expression of miR-221 and its potential target genes in human hepatocellular carcinoma cells / 中国中药杂志

To investigate the anti-hepatoma mechanism of α-pinene, HepG2 cell was treated with α-pinene and the change of cell cycle was examined by flow cytometry. The expression of miR-221, which was related the regulation of G₂/M phase, was detected by quantitative Real-time PCR. Meanwhile, TargetScan and other online bioinformatics methods were used to analyze and predict the target genes of miR-221, then the expression level of related target genes were detected by quantitative Real-time PCR. The results showed that α-pinene inhibited the proliferation of HepG2 cells in dose-dependent manner. It was also proved that HepG2 cells were arrested at G₂/M phase by α-pinene (P<0.05). The expression of miR-221 was down-regulated in α-pinene treated HepG2 cell. The bioinformatics analysis showed that CDKN1B/P27 and CDKN1C/P57 may be the protential targets of miR-221 and both of them were significantly up-regulated（P<0.001,P<0.05）by α-pinene treatment. According to these results, it was believed that α-pinene may inhibit the proliferation of hepatocellular carcinoma cells through arrest the cell at G₂/M phase, which may be associated with the down-regulate of the miR-221 expression and up-regulate of the CDKN1B/P27 and CDKN1C/P57 expression.

Protective effect of miR-221 onβcells in mice with diabetic nephropathy and its mechanism / 吉林大学学报(医学版)

Objective:To study the influence of miR-221 in theβcells of mice with diabetic nephropathy (DN), and to clarify the protective effect of miR-221 on DN.Methods:Twenty wild type C57BL/6J mice aged 8 weeks were divided into control group and DN group (n=10).The DN mice models were constructed with 14 weeks of high fat diet,and 100 mg·kg-1 streptozotocin (STZ)was used to induce the partial insulin deficiency.10 weeks later the blood glucose,serum creatinine,blood urea nitrogen content and 24h-urinary albumin excretion rate were detected.Theβcells of islet were isolated from the DN mice.The expression level of SOCS3 mRNA inβ cells of islet was valued by Real-time PCR.The proliferation of pancreaticβcells of islet was examined by MTT assay.The contents of insulin and insulin release levels in pancreaticβ cells of islet were detected by ELISA assay.Luciferase reporter gene assay was used to detect the luciferase reporter gene activity of SOCS3.Results:The DN mouse models were constructed successfully.The blood glucose,serum creatinine,blood urea nitrogen and 24h-urinary albumin excretion rate of the mice in DNA group were higher than those in control group (P < 0.05 ).After overexpression of miR-221,the expression level of SOCS3 mRNA in pancreaticβcells of islet and the proliferation ability ofβcells of islet of the mice in DN group were lower than those of normal islet cells (P < 0.05).Then luciferase reporter gene method found SOCS3 as one of the target genes of miR-221.After overexpression of miR-221,the insulin release level and insulin content in pancreaticβcells of islet were higher than those of normal islet cells (P <0.05).Conclusion:miR-221 can promopt the synthesis and secretion ofβ cells of islet and inhibit the dysfunction ofβcells of islet in the DN mice by down-regulating the SOCS3 level.

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.

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.

Suppression of miR-221 inhibits glioma cells proliferation and invasion via targeting SEMA3B

BACKGROUND: Gliomas are the most common primary tumors in the central nervous system. Due to complicated signaling pathways involved in glioma progression, effective targets for treatment and biomarkers for prognosis prediction are still scant. RESULTS: In this study we revealed that a new microRNA (miR), the miR-221, was highly expressed in the glioma cells, and suppression of miR-221 resulted in decreased cellular proliferation, migration, and invasion in glioma cells. Mechanistic experiments validated that miR-221 participates in regulating glioma cells proliferation and invasion via suppression of a direct target gene, the Semaphorin 3B (SEMA3B). The rescue experiment with miR-221 and SEMA3B both knockdown results in significant reversion of miR-221 induced phenotypes. CONCLUSION: Taken together, our findings highlight an unappreciated role for miR-221 and SEMA3B in glioma.