Predictive value of plasma microRNA-126 for treatment response of psoriasis patients to combined treatment with acitretin, methotrexate and ultraviolet phototherapy / 第二军医大学学报

Objective: To investigate the plasma microRNA-126 (miRNA-126) level in psoriasis patients before and after acitretin, methotrexate and ultraviolet phototherapy-combined treatment, and to assess its value in predicting the treatment outcomes. Methods: A total of 196 patients with moderate-to-severe psoriasis were consecutively enrolled and received combined treatment with acitretin, methotrexate and ultraviolet phototherapy, and 200 volunteers were recruited as healthy controls. Plasma samples of psoriasis patients were collected and miRNA-126 level was detected at baseline and after treatment for 1 month, 3 months and 6 months. Psoriasis area and severity index (PASI) were used to assess the disease severity. Treatment response was determined by PASI 50 response (PASI decreased by 50% compared with the baseline) and PASI 75 response (PASI decreased by 75% compared with the baseline). Spearman correlation test was used to analyze the correlation between the expression level of plasma miRNA-126 and PASI. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of relative expression of miRNA-126 in psoriasis patients. Univariate and multivariate logistic regression models were used to analyze the influencing factors of PASI 50 and PASI 75 responses. Results: Baseline miRNA-126 expression was significantly lower in psoriasis patients compared with healthy controls and was negatively correlated with PASI score (r= - 0.222, P=0.002). ROC curve displayed that miRNA-126 had a good diagnostic value for psoriasis (area under curve: 0.700). After treatment for 1 month, 3 months and 6 months, miRNA-126 levels were significantly elevated compared with the baseline (P<0.01). The response rates of PASI 50 were 7.1% (14/196), 37.2% (73/196) and 64.8% (127/196) after treatment for 1 month, 3 months and 6 months, and those of PASI 75 were 1.5% (3/196), 14.3% (28/196) and 35.7% (70/196), respectively. PASI 50 and PASI 75 responders presented lower baseline miRNA-126 level (P<0.05). Logistic regression analysis revealed that baseline miRNA-126 level was negatively associated with PASI 50 and PASI 75 responses. Conclusion: Plasma miRNA-126 level gradually increases in psoriasis patients after acitretin, methotrexate and ultraviolet phototherapy-combined treatment, and the baseline miRNA-126 level is negatively correlated with treatment response.

Role of microRNA-126-5p in myocardial injury induced by doxorubicin / 中国病理生理杂志

AIM:To observe the expression of microRNA-126-5p during myocardial injury and its role in myo-cardial cell injury induced by adriamycin(also called doxorubicin, DOX).METHODS: The BALB/c mouse model of DOX-induced acute and chronic myocardial injury was established via intraperitoneal injection of DOX.HE staining was applied to observe the morphological changes of myocardial tissues.Lactate dehydrogenase(LDH)in serum was detected and PowerLab system was used to detect the influence of DOX on the changes of ±dp/dtmax.The expression of microRNA-126-5p in injured myocardial tissues and the H 9c2 cells exposed to DOX was detected by real-time PCR.Gain-and loss-of-function experiments were conducted to detect the role of microRNA-126-5p in H9c2 cells treated with DOX on LDH release and caspase-3 activation.RESULTS:In acute and chronic DOX myocardial damage models in mice,HE staining showed disarranged myocardial fibers, dissolved myofibril and inflammatory cell infiltration.Higher serum LDH level and lower ±dp/dtmaxin DOX-treated mice than those in normal mice were found.Compared with the normal mice, the expression level of microRNA-126-5p was significant increased in the myocardium with DOX-induced injury.Similarly,the expression level of microRNA-126-5p was significant increased in the H9c2 cells treated with DOX.In addition, over-expression of microRNA-126-5p decreased cell viability and promoted apoptosis,while microRNA-126-5p ablation promoted the viability and inhibited the apoptosis of H9c2 cells.CONCLUSION:The microRNA-126-5p expression is up-regulated in myocar-dial injury induced by DOX,and microRNA-126-5p inhibits cell viability and promotes apoptosis induced by DOX.

Expression of microRNA-126 in Colon Cancer and Its Clinical Significance / 现代检验医学杂志

Objective To investigate the expression and biological significance of microRNA-126 in colon cancer tissue.Methods The expression of microRNA-126 in colon cancer tissues and adjacent tissues of colon cancer patients was detected by real-time quantitative polymerase chain reaction (RT-PCR) in 104 patients with colon cancer surgery (total of 104 pairs).The lentiviral vector was used to construct microRNA-126 cell line,and the effects of microRNA-126 on proliferation,migration and invasion of colon cancer cells were further studied in vitro.Results The relative expression of microRNA-126 in colon cancer tissues (0.63±0.11) was significantly lower than that in adjacent tissues (1.08±0.15),the difference was statistically significant(t=14.561,P＜0.01).The positive rate of microRNA-126 expression in colon cancer tissues (61.5％) was significantly lower than that in adjacent cancer tissues (86.5 ％) the difference was statistically significant(x2=16.908,P＜0.05).The expression of microRNA-126 was significantly correlated with Dukes stage,depth of invasion,lymph node metastasis and distant metastasis (P＜0.05).In the Transwell experiment with matrix glue,the number of cell migration in transfection group (11.26±4.85) was significantly lower than that in blank control group (264.37±32.15),the difference was statistically significant (t=23.418,P＜0.01).In the Transwell experiment without matrix glue,the number of cell migration in transfection group (83.75 ± 13.74) was significantly lower than that in blank control group (339.64 ± 26.38),the difference was statistically significant(t=12.682,P＜0.01).MicroRNA-126 could inhibit the proliferation and invasion of SW480 cells.Conclusion MicroRNA-126 can significantly inhibit the development of colon cancer cells and affect the biological behavior of colon cancer cells.

Relationship between microRNA-126 and curative effect of chemoradiotherapy in patients with gastric cancer, and enhancement effect of microRNA-126 on radiosensitivity of SGC-7901 cells / 中国病理生理杂志

AIM: To investigate the influences of microRNA-126 on the curative effect of chemoradiotherapy and radiosensitivity of SGC-7901 cells.METHODS: The patients of gastric cancer (n=60) were selected in this study including 32 males and 28 females with the average age of (51±7) years.All patients received similar chemoradiotherapy strategy.The tissue and blood samples were collected during treatment.The short-term curative effect was evaluated by the Response Evaluation Criteria in Solid Tumors (RECIST), and the patients were divided into sensitive group and insensitive group.The microRNA-126 levels were detected by RT-qPCR.The SGC-7901 cells were maintained in vitro and transfected with microRNA-126 mimic.The plate colony formation assay was used to determine the enhancement effect of microRNA-126 on radiosensitivity of the SGC-7901 cells.The apoptotic rate of the SGC-7901 cells induced by microRNA-126 was analyzed by flow cytometry.RESULTS: According to the RECIST, 28 cases were defined as sensitive patients and 32 cases were the insensitive patients.Compared with the sensitive patients, the microRNA-126 levels both in blood and tissue samples were lowered in the insensitive patients, and the relative fold changes were 0.72±0.04 and 0.48±0.03, respectively (P<0.05).After transfection with microRNA-126 minic, the SF2 and D0 in the SGC-7901 cells were decreased with the SER of 1.74.Furthermore, microRNA-126 induced apoptosis of SGC-7901 cells and enhanced their radiosensitivity.CONCLUSION: The patients with low microRNA-126 level may suffer a poor curative effect of chemoradiotherapy on the gastric cancer.MicroRNA-126 has an enhancement effect on the radiosensitivity to the SGC-7901 cells.

miRNA-126 inhibits proliferation, migration and invasion of human lung cancer A549 cells via EGFR/AKT/mTOR pathway / 中国病理生理杂志

AIM:To investigate the effects of microRNA ( miRNA)-126 on the proliferation , migration and in-vasion of human lung cancer cell lines , and to explore its mechanism .METHODS:The A549 cells were transfected with miRNA-126 agomir by Lipofectamine 2000.The expression of miRNA-126 was detected by real-time PCR.The cell activity was detected by MTT assay .The number of viable A549 cells was counted by the method of Trypan blue exclusion .The cell colony-forming capability was determined by cell colony formation test .The cell migration and invasion abilities were assayed by wound healing and Transwell methods , respectively.The protein levels of p-EGFR, EGFR, p-AKT, AKT, p-mTOR and mTOR were determined by Western blot .RESULTS:The expression level of miRNA-126 was significantly in-creased in the A549 cells compared with negative control ( NC) group and control group ( P<0.01 ) .The proliferation of A549 cells was decreased extremely after transfected with the miRNA-126 agomir (P<0.01), so did the result of the cell colony-formation test.The migration and invasion abilities of the lung cancer cells were also significantly inhibited .The protein levels of p-EGFR, p-AKT and p-mTOR were significantly down-regulated compared with NC group and control group ( P<0.01) .CONCLUSION:Over-expression of miRNA-126 significantly inhibits the proliferation , migration and invasion ability of human lung cancer A 549 cells by down-regulation of EGFR/AKT/mTOR pathway .

Biological significance of miR-126 expression in atrial fibrillation and heart failure

We investigated the biological significance of microRNA-126 (miR-126) expression in patients with atrial fibrillation (AF) and/or heart failure (HF) to examine the possible mechanism of miR-126-dependent AF and development of HF. A total of 103 patients were divided into three groups: AF group (18 men and 17 women, mean age: 65.62±12.72 years), HF group (17 men and 15 women, mean age: 63.95±19.71 years), and HF-AF group (20 men and 16 women, mean age: 66.56±14.37 years). Quantitative real-time PCR was used to measure relative miR-126 expression as calculated by the 2−ΔΔCt method. miR-126 was frequently downregulated in the 3 patient groups compared with controls. This reduction was significantly lower in permanent and persistent AF patients than in those with paroxysmal AF (P<0.05, t-test). Moreover, miR-126 expression was markedly lower in the HF-AF group compared with the AF and HF groups. The 3 patient groups had higher N-terminal prohormone brain natriuretic peptide (NT-proBNP) levels, lower left ventricular ejection fraction (LVEF), larger left atrial diameter, and higher cardiothoracic ratio compared with controls. There were significant differences in NT-proBNP levels and LVEF among the AF, HF, and HF-AF groups. Pearson correlation analysis showed that relative miR-126 expression was positively associated with LVEF, logarithm of NT-proBNP, left atrial diameter, cardiothoracic ratio, and age in HF-AF patients. Multiple linear regression analysis showed that miR-126 expression was positively correlated with LVEF, but negatively correlated with the logarithm of NT-pro BNP and the cardiothoracic ratio (all P<0.05). Serum miR-126 levels could serve as a potential candidate biomarker for evaluating the severity of AF and HF. However, to confirm these results, future studies with a larger and diverse patient population are necessary.

MicroRNA-126 inhibits the proliferation of lung cancer cell line A549

Objective: To study the role of microRNA-126 in the development of lung cancer. Methods: The biological function of microRNA-126 was detected using EdU assay and CCK-8 assay; the target gene of microRNA-126 was analyzed using real time RT-PCR and Western blot assay. Results: In A549 cell line, overexpression of microRNA-126 inhibits the proliferation rate; VEGF is the target gene of microRNA-126; microRNA-126 exerts its function via regulating VEGF protein level. Conclusions: microRNA-126 inhibits the proliferation in A549 cell line.

MicroRNA-126 inhibits the proliferation of lung cancer cell line A549

Objective:To study the role of microRNA-126 in the development of lung cancer.Methods:The biological function of microRNA-126 was detected using EdU assay and CCK-8 assay;the target gene of microRNA-126 was analyzed using real time RT-PCR and Western blot assay.Results: In A549 cell line, overexpression of microRNA-126 inhibits the proliferation rate; VEGF is the target gene of microRNA-126; microRNA-126 exerts its function via regulating VEGF protein level.Conclusions: microRNA-126 inhibits the proliferation in A549 cell line.

Effect of microRNA126 on glucose metabolism in the normal liver cell lines / 中华内分泌代谢杂志

[Summary] To investigate the effect of microRNA126 on glucose metabolism in the normal liver cell lines. In vitro, the chang liver cell lines were cultured. Under the most effective transfection conditions ascertained above, microRNA126 mimic, microRNA126 inhibitor, and relative negative control were transfected into the cultured normal liver cells. And the transfection efficiency was tested by realtime fluorescent quantitative PCR. After 48 hours, the cells were stimulated with synthetic insulin ( 100 nmol/L ) and respective substrates for 2 hours. Then the glycogenesis, gluconeogenesis, and glycolysis in cells were measured. The level of microRNA126 of the microRNA126 mimic group was higher than the other groups, and the difference was statistically significant ( P<0. 05 ). MicroRNA126 mimic group significantly decreased glucose utilization, reduced glycogen synthesis, effectively increased the account of gluconeogenesis, reduced lactate production, and pyruvate kinase activity ( all P<0. 05). The over-expressing microRNA126 in hepatocytes may reverse the function of glucose metabolism, and enhance output of hepatic glucose.

MicroRNA-126 inhibits the proliferation of lung cancer cell line A549

OBJECTIVE@#To study the role of microRNA-126 in the development of lung cancer.@*METHODS@#The biological function of microRNA-126 was detected using EdU assay and CCK-8 assay; the target gene of microRNA-126 was analyzed using real time RT-PCR and Western blot assay.@*RESULTS@#In A549 cell line, overexpression of microRNA-126 inhibits the proliferation rate; VEGF is the target gene of microRNA-126; microRNA-126 exerts its function via regulating VEGF protein level.@*CONCLUSIONS@#microRNA-126 inhibits the proliferation in A549 cell line.

Expressions of plasma microRNA-126 and microRNA-1 in children with asthma exacerbation and its clinical significance / 中华实用儿科临床杂志

Objective To investigate the changes in plasma microRNA-126 and microRNA-1 in children with asthma exacerbation and its relationship with bronchial asthma.Methods From October 2012 to December 2013,48 children with asthma exacerbation from the Outpatient Department and the Inpatient Department in Houjie Hospital Affiliated to Guangdong Medical College were enrolled in the study (asthma group).Meanwhile,52 healthy children wcre selected as the healthy control group.The expression levels of plasma microRNA-126 and microRNA-1 were detected by real-time quantitative PCR (RT-PCR).The content of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in plasma was measured by enzyme-linked immunosorbent assay (ELISA).The predictive value of microRNA-126 and microRNA-1 in plasma to bronchial asthma was evaluated by receiver operating characteristic (ROC) curve.Results The relative expression levels of plasma microRNA-126 in the asthma group were upregulated compared with those in the healthy control group [7.36 (0.96-41.21) vs 3.68 (0.75-38.91),Z =3.135,P =0.038],and microRNA-1 relative expression levels in the asthma group were lower than those of the healthy control group [2.17 (0.18-26.97) vs 5.83 (0.82-39.62),Z =2.156,P =0.045].The content of IL-4 in asthma group was higher than those of the control group [(109.98 ± 74.58) ng/L vs (78.50 ± 75.82) ng/L,t =2.122,P =0.036],and the IFN-γ level in the asthma group was lower than those of the healthy control group [(70.49 ± 12.03) ng/L vs (77.03 ± 17.16) ng/L,t =2.270,P =0.025].In the plasma of patients with asthma exacerbation,the sensitivity of microRNA-126 and microRNA-1 was 85.42％ (41/48 cases)and 79.17％ (38/48 cases),respectively.The specificity of microRNA-126 and microRNA-1 in healthy controls was 78.85％ (41/52 cases) and 73.08％ (38/52 cases),respectively.The area under ROC curve of microRNA-126 and microRNA-1 was 0.919 (95％ CI 0.866-0.973),0.867 (95％ CI 0.796-0.939).Conclusions MicroRNA-126 is significantly elevated in plasma of children with asthma exacerbation.The plasma levels of microRNA-1 were significantly downregulated.These results suggest that microRNA-126 and microRNA-1 may be potential markers for the diagnosis of bronchial asthma.

Purification and identification of bone marrow mesenchymal stem cells and microRNA126 expression during their differentiation into vascular endothelial cells / 第二军医大学学报

Objective To establish a method for isolation, purification of bone marrow mesenchymal stem cells (MSCs) and for their differentiation into vascular endothelial cells, and to observe the regulatory role of microRNA126 expression during the differentiation. Methods MSCs were isolated from normal rat bone marrow using gradientdensity centrifugation and repeated attachment method. CD34, CD105and CD73 expressions in MSCs were detected with immunofluorescent staining. MSCs were cultured with MEF medium for differentiating into endothelial cells; CD34, VE-cadherin and microRNA126 expressions were examined by qRT-PCR at different time points. Results Immunofluorescent detection demonstrated that MSCs have been isolated and purified successfully, with MSCs negative for CD34, strongly positive for CD105 and positive for CD73. The purified MSCs had a good uniformity and a purity above 90%. qRT-PCR examination revealed that CD34 and VE- cadherin expressions were not detected on day 1-4 of induction, strongly positive on day 5-6, and on day 7-9, VE-cadherin was still positive, CD34 decreased on day 7, and increased again on day 8-9. Interestingly, the expression of CD34 and microRNA126 mRNA was consistent during the differentiation. Conclusion We have successfully established a method for MSCs isolation and differentiation into endothelial cells; microRNA126 may play a regulatory role in MSCs differentiation into endothelial cells.

Expressions and clinical significances of microRNA-126 and microRNA-7 in esophageal squamous cell carcinoma / 国际肿瘤学杂志

Objective To detect the expressions of microRNA-126 (miR-126) and microRNA-7 (miR-7) in esophageal squamous cell carcinoma (ESCC) and to analyze their correlations with clinicopathologic features and prognosis of ESCC.Methods The expressions of miR-126 and miR-7 in 116 ESCC samples and matched normal tissue samples were detected by real-time PCR.Statistical analysis was used to find the relationships among the expressions of miR-126 and miR-7,pathological characteristics and prognosis.Results Low expression,normal expression and high expression of miR-126 were found in 73 (62.9％),35 (30.2％) and 8 (6.9％) carcinoma samples respectively.Low expression,normal expression and high expression of miR-7 were found in 52 (44.8％),35 (30.2％) and 29 (25.0％) carcinoma samples respectively.The disease-free survival in patients with low expression of miR-126 and miR-7 was shorter than that in patients with non-low expression (x2 =4.268,P ＜0.05 ; x2 =4.993,P ＜0.05).The low expression of miR-126 was correlated with tumor location,family history and drinking (x2 =14.564,P ＜ 0.05 ; x2 =5.691,P ＜ 0.05 ; x2 =4.971,P ＜ 0.05),but was uncorrelated with gender,age,diferentiation,infiltration,lymphatic metastasis and smoking (all P ＞ 0.05).The low expression of miR-7 was uncorrelated with pathological characteristics of ESCC (all P ＞ 0.05).Conclusion The low expressions of miR-126 and miR-7 may be related to the prognosis of patients with ESCC,and have a certain clinical detection significance.