miR-155 Research Articles

Expression and function mechanism of microRNA-126 in esophageal squamous cell carcinoma

Objective To investigated microRNA(miR)-126 expression and its potential targets in esophageal squamous cell carcinoma(ESCC). Methods The expression level of miR-126 was detected in cancerous and paired paracancerous tissues from 75 patients with ESCC, association between miR-126 level and clinicopathologic characteristics were analysised.Target analysis of miR-126 was predicted using online tools.The effect of miR-126 expression on target proteins was assessed through up/ down miR-126 level in ESCC cell lines. Results The relative expression of miR-126 in ESCC tissues was 0. 28±0. 32 and 0. 45±0. 47 in normal tissues(P< 0. 01).Low level of miR-126 was associated with tumor differentiation, lymph nodes metastasis, tumor in-depth, and TNM stages (P< 0. 05).Insulin receptor substrate-1(IRS-1) was over-expressed and associated with cell differentiation(P< 0. 01).In ESCC cell line Eca9706, Eca109 and TE-1, miR-126 mimics down-regulated the expression of IRS-1 protein(0. 785±0. 337, 1. 873±0. 684, 1. 938±1. 081) compared with non-treated control(1. 188±0. 336, 2. 756±1. 097,3. 028±0. 789, P< 0. 01), whereas miR-126 inhibitors lead to the opposite results(2. 543±0. 610, 5. 182±1. 897, 5. 940±0. 997, P<0. 01). Conclusion This study revealed that miR-126 is low-expressed in ESCC and acts as a tumor suppressor in the carcinogenesis of ESCC.IRS-1 was downstream targets of miR-126 in ESCC. Key words: Esophageal squamous cell carcinoma; MicroRNA-126; Insulin receptor substrate-1

Correlation between microRNA-155 and WEE1 expression and their role in colon cancer

Objective To investigate the role of microRNA(miR)-155 and WEE1 in the tumorigenesis of colon cancer by detecting their expression level in tumor and normal tissue. Methods The expression level of miR-155 and WEE1 from tumor and normal tissues of 187 patients were detected by real-time fluorescent quantitative polymerase chain reaction(FQ-PCR).Correlation between variables was analyzed by Kendall'tau method.Survival curves were plotted using Kaplan-Meier method with Log-rank test.Cyclooxygenase(COX) proportional hazard model was used to evaluate the independent prognostic factors. Results miR-155 expression negatively and statistically significantly correlates with WEE1 expression in colon cancer tissue(r=-0. 555, P< 0. 01). miR-155 and WEE1 expression level was closely related with differentiation, TNM stage and lymphovascular metastasis(P< 0. 01).miR-155 and WEE1 expression level, lymphovascular metastasis were independent prognostic factors in colon cancer patients.The median overall survival difference between patients with increased level of miR-155 expression and normal levels of miR-155 expression had statistical significance(23 months vs.85 months,P<0. 05).The median overall survival difference between patients with decreased level of WEE1 expression and normal levels of WEE1 expression had statistical significance(25 months vs.70 months, P< 0. 05). Conclusion miR-155 and WEE1 expression level were independent prognostic factors of colon cancer patients. Decreased miR-155 expression and increased WEE1 expression predict a good prognosis. Key words: MicroRNA-155; WEE1; Colon cancer; Real-time polymerase chain reaction

Transcriptomics of psoriasis

Psoriasis is a chronic inflammatory multigenic dermatosis of unclear etiology. Recently, more and more studies have been carried out to investigate into its physiopathologic mechanism at transcriptome level. The methods used in these studies mainly include expressed sequence tag(EST)analysis, GeneChip, second-generation sequencing technology, etc. Some non-coding RNAs, including miR-21, miR-203 and miR-31, have been found to be expressed abnormally in psoriatic lesions, and closely related to the differentiation and proliferation of keratinocytes. These findings provide important evidences for the pathogenesis of psoriasis. Key words: Psoriasis; Microchip analytical procedures; Multilocus sequence typing

The differential expression profiling of microRNAs in cutaneous wounds between diabetic rats and nor-mal rats

Objective To explore the differential expression profiling of microRNAs(miRNAs)in cutaneous wounds between diabetic rats and normal rats, and further investigate its significance in wound healing. Methods The diabetic rat models were induced by intraperitoneal injection of 65 mg/kg of streptozotocin(STZ); after four weeks, create full-thickness excisional dorsal wounds in 6 established diabetic model rats and normal rats respectively; then wound tissues were harvested on day 3.Use Trizol to extract total RNA from the sample; purify it after quality inspection qualified; then to the procedures of sample labeling and hybridization. Now hybrid images can be obtained through scanning the washed microarray slides by Agilent Scanner. Analyze these images and extract the data by agilent feature extraction software; then normalize and analyze the data by agilent genespring software. The miRNA profiling results were validated by real- time reverse transcriptase polymerase chain reaction(Real-time PCR). Results A total of 83 miRNAs were differentially expressed in cutaneous wounds of diabetic rats compared with normal rats, among which 18 were up-regulated while 65 are down-regulated. The significantly up-regulated miRNAs were rno-miR-496-5p, rno-miR-105, rno-miR-122-5p,etc;and the significantly down-regulated miRNAs were rno-miR-196a-5p, rno-miR-134-5p, rno-miR-31-3p, etc.The results of Real-time PCR have good consistency with the microarray results.KEGG pathways enrichment analysis shows that miR-31, miR- 222, miR-449a and other significant differential expressed miRNAs were respectively involved in mitogen-activated protein kinase(MAPK), Wnt, Notch and other signaling pathways which link to wound healing. Conclusion There was significant differential miRNAs expression in cutaneous wounds between diabetic rats and normal rats, which may be closely related to the mechanisms of diabetic wound healing. Key words: Diabetes mellitus; Wound healing; MicroRNAs; Differential expression

Preliminary study of impact of steroids on expression profile and transcriptome of bone microvascular endothelial cells

To explore the differential expression profiles and transcriptomes of human bone microvascular endothelial cells via microarray and elucidate the molecular mechanisms underlying steroids-induced osteonecrosis. Bone microvascular endothelial cells in cancellous bone of femoral head were harvested by type I collagenase and trypsin digestion and identified by immunofluorescent labeling with primary antibodies specific for vascular endothelial cells' biomarkers such as von Willebrand factor, platelet endothelial cell adhesion molecule 1, vascular endothelial cadherin and vascular cell adhesion molecule 1. Their differential expression profiles and transcriptomes were tested by double-labeled lncRNAs + mRNAs microarrays and single-labeledd microRNAs microarrays. The data were analyzed by bioinformatical software with database checking. The authors spatial correlations and expression relevance were established among lncRNAs, microRNAs and mRNAs. Steroids incurred specific changes in mRNAs, microRNAs and lncRNAs transcribed in human bone microvascular endothelial cells of femoral head in vitro cultured. Thus a model of coexpression network was constructed among transcript modules with bioinformatical tools such as target gene prediction. Steroids induced significant changes in transcripts of bone microvascular endothelial cells such as miR-100, miR-222, miR-933, miR-339 and miR-23. They exert considerable influences on osseous tissues and circulatory systems in cancellous bone by targeting cytokines and enzymes such catalase, fibroblast growth factor and nerve growth factor. Through further explorations of these transcripts, we can not only elucidate the underlying pathogenesis, but also discover specific biomarkers and therapeutic targets for corresponding diseases.

Cell proliferation and invasion in microRNA-143 and microRNA-29a transfected bladder cancer cells and its relationship of ras association domain family 1A methylation

Objective To evaluate the relationship among bladder cancer cell proliferation and invasion,microRNA-143 and 29a,and ras association domain family 1A (RASSF1A) methylation.Methods MiR-143 and miR-29a mimics were transfected to T24 cell lines.Transfection efficiency was confirmed by reverse transcription polymerase chain reaction (RT-PCR).All cell lines were divided into four groups:WT group (wild T24 cell lines without any treatment),NC group (transfected by negative control mimics),M1 group (transfected by miR-143 mimics) and M2 group (transfected by miR-29a mimics).Apoptosis rates,cell cycle,cell invasive inhibitor rates,RASSF1 A gene expression and methylation were detected.Results Transfection was successful confirmed by RT-PCR.Apoptosis rates of WT,NC,M1 and M2 group were 2.5％,3.4％,12.4％ and 13.3％ respectively.S-phase cells in M1 and M2 group increased significantly.Cell invasive inhibitor rates of NC,M1 and M2 group were 2.0％,16.5％ and 13.2％ respectively which showed the rates in M1 and M2 group were higher than NC group (P ＜0.05).RASSF1A gene expression was upgraded in M2 group and no changes were found in M1 group.What's more,MSP results showed that RASSF1 A gene was partial methylated in M2 group while it was completely methylated in other groups.Conclusion T24 bladder cancer cell proliferation and invasion decreased after miR-143 and miR-29a transfected.miR-29a transfection may inhibit RASSF1 A gene methylation and lead to gene expression uploaded. Key words: MicroRNA; Bladder cancer; Ras association domain family 1A; Methylation; Transfection

Study on the levels of microRNA-146b, microRNA-155 and microRNA-30b in human milk and their correlation with maternal and infant health

Objective To explore the expression levels of immune-related microRNA-146b(miR-146b), microRNA-155(miR-155) and microRNA-30b(miR-30b) in human breast milk and its relationship with maternal and infant's health. Methods One hundred and thirty-four mothers and their infants from obstetrical department were recruited in the study after delivery.The subjects were divided into 2 groups, breast feeding group(n=86) and formula-fee-ding group(n=48), and were followed up 3 months after delivery.Breast milk samples were collected at 2-5 days after delivery(colostrum) and 3 months after delivery(mature milk). The expression levels of microRNAs in milk samples were detected by real-time PCR.The relationship between levels of microRNAs and maternal and infant-related factors was analyzed. Results 1.MiR-146b, miR-155 and miR-30b expressions were abundant both in human colostrums(5.950±0.823, 3.899±0.920, 4.057±0.604) and mature milk(4.840±0.805, 2.128±0.969, 4.929±0.566). The levels of miR-146b and miR-155 were higher in colostrum than that of mature milk(t=7.716, 10.215, all P<0.01), while the level of miR-30b was higher in mature milk than that of colostrums(t=-8.626, P<0.01).2.Additionally, the level of miR-30b was negatively correlated with maternal pre-pregnancy body mass index(r=-0.298, P<0.01).3.The levels of miR-146b and miR-30b were higher in mothers giving birth by vaginal delivery than those who underwent cesarean section(t=2.356, 3.108, all P<0.05).4.The levels of miR-146b and miR-155 were higher in colostrum-fed girls than boys(t=-2.204, -2.985, all P<0.05).5.The level of miR-146b in mature milk was negatively correlated with 3-month-old infant's Z score of body weight(r=-0.425, P<0.05) and body length(r=-0.569, P<0.01).6.During follow-up, the incidence of baby eczema in breast feeding group(8.82%, 3/34 cases) was lower than that in formula milk feeding group(29.17%, 14/48 cases) (χ2 = 5.012, P=0.025). Conclusions The levels of immunocompetent microRNAs in human milk are influenced by the lactation period, maternal pre-pregnancy body mass index, mode of delivery and infant sex.The immune-related microRNAs in human milk could be involved in the regulation of infant's immunity and growth. Key words: Human milk; Immunity; MicroRNA-146b; MicroRNA-155; MicroRNA-30b; Maternal and infant health

Expression and effect of microRNA-214 in advanced hypopharyngeal carcinoma

To investigate the expression of microRNA-214(miR-214) in advanced hypopharyngeal carcinoma tissues and its effects on the invasion, migration and colone formation of FaDu cells. miR-214 expression in 30 cases of advanced hypopharyngeal carcinoma tissues and normal hypopharyngeal mucosa tissues was detected by real-time quantitative polymerase chain reaction (RT-qPCR). miR-214 was upregulated through transfecting the overexpression vector hsa-mir-214 into FaDu cells. The influences of miR-214 upregulation on the invasion, migration, clone formation and Twist expression were measured by Transwell invasion, Transwell migration, plate clone formation and Western blot assays, respectively. The expression of miR-214 in advanced hypopharyngeal carcinoma tissues (0.311 ± 0.206) was significantly less than normal hypopharyngeal mucosa tissues (1.620 ± 1.394; t = 5.09, P < 0.05) . The expression of miR-214 was notably upregulated after tranfected with hsa-mir-214 compared with the negative control group (t = 6.347, P < 0.05). The migration and invasion ability of FaDu cells transfeced with hsa-mir-214 was decreased by comparison with negative control cells (t = 11.6, P < 0.01; t = 6.499, P < 0.05). There was no significant difference of the average clony number and the cloning efficiency between the experimental and negative control groups (t = 0.592, P > 0.05). of Western blot assay showed that, Twist expression in the miR-214-overexpressed group was apparently decreased compared with that in the control group (t = 6.545, P < 0.05). miR-214 is expressed at a low level in advanced hypopharyngeal carcinoma tissues, and can obviously inhibit the invasion and migration abilities of FaDu cells, possibly because of its inhibiting effect on Twist expression. Additionally, miR-214 plays no significant role in the proliferation of FaDu cells.

Melatonin regulating the expression of miRNAs involved in hair follicle cycle of cashmere goats skin.

Melatonin and microRNAs (miRNAs) play important roles in regulating hair follicle development. However, the effect of melatonin on the expression pattern of miRNAs in skin and follicle of cashmere goats remain largely undefined. To explore the mechanism of melatonin affecting cashmere growth mediated by miRNAs, the effect of melatonin implants administered in Nei Mongol cashmere goats was assessed. In the experiment, five yearling does were implanted with melatonin, with the remaining other five females as control group. The expression of six candidate miRNAs was quantified by reverse transcription-real time polymerase chain reaction (RT-qPCR). The results indicated that melatonin significantly altered the expression pattern of miRNAs. Except for let-7a, the expression levels of miR-203, miR-205, miR-96, miR-183 and miR-199a occur three transitions during a cashmere cycle; melatonin changed the co-expression pattern of miRNAs. The correlation coefficient between miRNAs is 0.87-0.99 in control group(P<0.01). Compared with the control group, the correlation coefficient between some miRNAs (let-7a and miR-96, miR-199a, miR-205;miR-203 and miR-96, miR-199a; miR-96 and miR-183; miR-183 and miR-199a) was significantly weakened in melatonin group. Melatonin might induce second growth of cashmere mediated by down-regulating the expression level of some miRNAs in June in melatonin implanted group.

Genomics analysis of miRNA in colon cancer tissues

To examine the differential expression of miRNAs in colon cancer tissues and matched tumor adjacent tissues. Differential expression of microRNAs in twenty paired human colon cancer tissues and adjacent tissues were detected by QPCR. miRNAs with significantly differential expression (fold change >2.4 and P<0.01) were screened to analyze their accumulation by Cluster analysis. Thus correlation of miRNA and other colon cancer-associated proteins was examined. Expressions of 17 miRNAs were significantly reduced in colon cancer tissues. Cluster analysis showed that miR763-3, miR451 and miR99a had similar expression. Plasma CK20 level was negatively correlated with miR100 (r=-0.948), miR152a-5p (r=-0.948), miR125b (r=-0.949), miR145 (r=-0.949) and miR145*(r=-0.949) (all P<0.05). miR145 and other 16 miRNAs may be used as diagnostic molecular markers of colon cancer, and miR100, miR125a-5p, miR125b, miR145 and miR145* may become the molecular markers of colon cancer lymph node metastasis.

Role of AT1R-associated microRNAs in thyrotoxic heart desease

MicroRNAs (miRNAs) are small endogenous non-coding RNA molecules.They regulate negatively the expression of target genes at post-transcriptional stages by means of affecting the stability of target mRNA or interfering with the transcriptional process.Recently,there is evidence demonstrating that miRNAs play important roles in the gene expressions of thyrotoxic heart diseases.Elevated levels of thyroid hormones profoundly influence the cardiovascular system through the renin-angiotensin system (RAS).As a principal active component of RAS,angiotensin Ⅱ receptor 1 (AT1 R) interacts with miRNAs in promoting or extenuating the progress of thyrotoxic heart disease.In this article,the roles of AT1R-associated miRNAs,miR-21,miR-155,miR-208a/b,and miR-499 in thyrotoxic heart disease were reviewed. Key words: microRNA; Renin-angiotensin system; Angiotensin Ⅱ receptor 1 ; Thyrotoxic heart disease

Progress of microRNAs in pathogenesis of sepsis

MicroRNAs are small, single-stranded and non-coding RNAs.MicroRNAs modulates gene expression by binding themselves to complementary sites of target genes.It had been discovered that microRNAs modulate sepsis in multiple levels.Through targeting Ca2+ /calmodulin-dependent protein kinase Ⅱ, miR-152 and miR-148 impair innate response and antigen presentation, and also suppress proliferation of antigen specific CD4+ T cells; miR-29 family regulate cellular immunity through setting the threshold in thymic involution; MicroRNA-17-92 plays a causative role in B cell lineage; miR-146 and miR-155 down-regulate proinflammatory cytokines through suppressing components of pattern recognition receptors, while miR-132 through acetylcholinesterase degeneration.miR-124 degenerates mRNA of glucocorticoid receptorαand results in glucocorticoid resistant in treatment of sepsis.Immune regulation of microRNAs in sepsis is briefly described in this paper. Key words: MicroRNA; Sepsis; Immunomodulation

The expression of microRNAs in serum of patients with colorectal cancer and its clinical significance

Objective To explore the expression characteristic of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 in colorectal cancer, evaluate the clinical significance of candidate miRNAs in CRC diagnosis. Methods Case-control study was used. The expression levels of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 from 50 CRC patients and 27 normal controls were detected by real-time PCR, the levels of miRNAs expression in serum of 8 CRC patients 1 day before and 7 days after radical surgery were analyzed. The sensitivity and specificity of serum miRNAs expression for the diagnosis of colorectal cancer were evaluated using the receiver operating characteristic (ROC) curve. Results The expression level of mir-16, miR-21, mir-92a in serum of patients with CRC were (75.55±37.73), (35.96±23.81), (24.79±8.97) fmol/ml, significantly higher than that of the healthy control group (32.73±18.94), (24.36±13.27), (16.36±5.58) fmol/ml (tmir-16=2.77, tmir-21=2.34, tmir-92a=3.85,P 0.05). In addition, serum miR-16 and miR-92a levels of CRC patients 7 days after tumor resection were (36.02 ± 19.95), (14.82 ± 7.78) fmol/ml, significantly lower than that before operation (62.18 ± 34.17), (24.06 ± 12.99) fmol/ml (tmir-16=3.59, tmir-92a=2.60,P<0.05). Area under the ROC curve (AUC) of combined detection of mir-16, miR-21and mir-92a was 0.877, the sensitivity and specificity were 88% and 85%, respectively, which was higher than any of 3 miRNAs alone and the conventional tumor marker CEA. Conclusion Combination of mir-16, mir-21 and mir-92a in the diagnosis of CRC shows a better sensitivity and specificity, which would be expected as new tumor biomarkers for noninvasive diagnosis and monitoring progression of colorectal cancer. （Chin J Lab Med,2014,37:691-695） Key words: Colorectal neoplasms; microRNAs; Tumor markers, biological

Effect of smoking on the microRNAs expression in pneumoconiosis patients

To investigate the effect of smoking on the microRNAs (miRNAs) expression in pneumoconiosis patients. Real-time qPCR was used to measure the expression levels of miR-21, miR-200c, miR-16, miR-204, miR-206, miR-155, let-7g, miR-30b, and miR-192 in 36 non-smoking patients with pneumoconiosis and 38 smoking patients with pneumoconiosis, and the differences in expression levels between the two groups were evaluated by two-independent samples t-test. The expression of miR-192 in serum showed a significant difference between non-smoking and smoking pneumoconiosis patients (P < 0.05), and it decreased gradually in smoking patients with stage I and II pneumoconiosis. In the serum of all pneumoconiosis patients, the expression level of miR-16 was the highest, while the expression level of miR-204 was the lowest. Pneumoconiosis patients have differential expression of miRNAs in serum, and smoking has an effect on the miRNAs expression in pneumoconiosis patients.

Partial microRNA profiling differences between peripheral plasma and mononuclear leucocytes in schizophrenia patients

Objective To explore the differences of microRNA expression as specific blood biomarker between peripheral plasma and mononuclear leucocytes in schizophrenia patients. Methods A total of 60 schizophrenia patients and 72 healthy controls were enrolled into our study, and nine most reported schizophrenia-associated microRNAs (miR-30e, miR-34a, miR-181b, miR-195, miR-346, miR-432, miR-7, miR-132 and miR-212) expression levels were tested in peripheral plasma and mononuclear leucocytes using real-time PCR. Results MiRNA-132, miRNA-195, miRNA-30e and miRNA-7 (5.06±1.16, 2.99±1.27, 1.88±1.62, 9.59±1.85) showed significantly higher expression in plasma samples(5.91±0.93, 4.45±1.06, 3.56±1.30, 11.8±2.53)(P<0.05, P<0.001); miRNA-212, miRNA-34a and miRNA-30e(0.28±0.54, 2.69±1.10, -4.72±1.01) also showed significantly higher expression in mononuclear leucocytes samples (0.84±0.64, 3.97±1.01, -3.55±1.26)(P<0.05, P<0.001). ROC curve analysis suggested that the AUC of microRNA-30e in plasma was 0.767 (95%CI: 0.608~0.926) with sensitivity of 90.90% and specificity of 60.00%, and the AUC of microRNA-30e in mononuclear leucocytes was 0.756 (95%CI: 0.584-0.929) with sensitivity of 81.80% and specificity of 68.00%. Conclusion The microRNA expression in plasma varies greatly with that in mononuclear leucocytes. MicroRNA-30e can serve as specific blood-based biomarker for schizophrenia diagnosis. Key words: Schizophrenia; MicroRNA; Biomarker

DNA methyltrans-ferase 3b regulating the Cyclin D1 gene expression by microRNA-145 in the human hepatocellular carcinoma cell line

Objective To discuss the mechanism of the DNA methyltrans-ferase 3b (DNMT3b) regulating the Cyclin D1 gene expression in SMMC7721 cell line.Methods DNMT3b small interfering RNA (siRNA) was transfected into SMMC-7721 cells.Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of the microRNA (miR)-145 and Cyclin D1 mRNA.MSP was used to detect whether the promoter of miR-145 was methylated.Results DNMT3b has been successfully suppressed in the culture cells transfected by miR-145 with a transfection efficiency over 90％ ; The methylation status of miR-145 gene showed no significant differences between two groups (P ＞0.05) ; The expression of miR-145 was significantly higher than in control group (P ＜ 0.01) and the change between two groups had no difference (P ＞ 0.05).Conclusion DNMT3b can regulate the expression of Cyclin D1 by miR-145,affect the cell cycle and cause a large number of apoptosis of SMMC-7721 cells. Key words: Carcinoma, hepatocellular; DNA methyltrans-ferase 3b; Cyclin D1; Small interfering RNA; MicroRNA-145

Functions of miR126 and innate immune response.

MicroRNAs (miRNAs) are endogenous non-coding small RNAs that negatively regulate gene expression at the post-transcriptional level by base pairing with target mRNAs. Many miRNAs have been proven to regulate a broad range of processes, including proliferation, differentiation, development and apoptosis. miR126 is encoded by intron 7 of the EGF- like domain 7 (Egfl7) gene and highly expressed in human endothelial cells (ECs) and plasmacytoid dendritic cells (pDCs). Based on the current knowledge, miR126 is involved in angiogenesis and cancer. Also miR126 is the first miRNA which has been reported to involve in preconditioning the responsiveness of the host to pathogen infection in the steady state. This implicated that miR126 may be the potential therapeutic target for cancer and autoimmune diseases. In this review, we summarize the functions of miR126 in angiogenesis and cancer, and emphasize its relationship with innate immune response.

Role of spinal neuronal microRNA-132 in a mouse model of bone cancer pain

Objective To evaluate the role of spinal neuronal microRNA-132 (miR-132) in a mouse model of bone cancer pain (BCP).Methods Male C3H/HeJ mice,aged 4-6 weeks,weighing 20-25 g,were used in the study.The experiment was performed in 2 parts.In experiment Ⅰ,fifty-two male C3H/HeJ mice were randomly divided into 2 groups (n =26 each) using a random number table:sham operation group (S group) and BCP group.In experiment Ⅱ,thirty-two male C3H/HeJ mice were randomly divided into 4 groups (n =8 each) using a random number table:sham operation + intrathecal DNAse/RNAse-free water group (S + Veh group),BCP + intrathecal DNAse/RNAse-free water group (BCP + Veh group),BCP + intrathecal negative control locked nucleic acid (LNA) group (BCP + C group),and BCP + intrathecal miR-132 antisense LNA group (BCP + AS group).BCP was induced by injecting 2 x 105 NCTC 2472 cells into the distal medullary cavity of the right tibia.In BCP+ C and BCP + AS groups,negative control LNA and miR-132 antisense LNA 12 pmol/5 μl were intrathecally injected,respectively,once a day for 7 consecutive days starting from day 14 after inoculation.In S + Veh and BCP + Veh groups,DNAse/RNAse-free water 5 μl was intrathecally injected once a day for 7 consecutive days starting frowm day 14 after inoculation.In S and BCP groups,the paw withdrawal threshold to mechanical stimuli (MWT) and the number of spontaneous flinches (NSF) were measured,and the expression of miR-132 in spinal cord was detected before inoculation and on 4,7,10,14 and 21 days after inoculation.In S + Veh,BCP + Veh,BCP + C and BCP + AS groups,the parameters mentioned above were measured at 14 days before inoculation and 15,16,17,18,19,20 and 21 days after inoculation.Results Compared with S group,MWT was significantly decreased,NSF was increased and the expression of miR-132 was up-regulated on 7-21 days after inoculation in BCP group.Compared with S + Veh group,MWT was significantly decreased,and NSF was increased at each time point in the other three groups.Compared with BCP + Veh group,MWT was significantly increased,and NSF was decreased on 16-21 days after inoculation in BCP + AS group.Conclusion Spinal neuronal miR-132 is involved in the development and maintenance of BCP in mice. Key words: MicroRNAs ; Spinal cord ; Bone neoplasms ; Pain

Effects of KLF2 on ox-LDL induced expression of microRNA-146a and proinflammatory cytokines in human umbilical vein endothelial cells

Objective To investigate the effects of Kr uppel-like factor 2 （ KLF2 ） on oxidized low density lipoprotein （ox-LDL） induced expression of microRNA-146a （miR-146a） and proinflammatory cyto-kines （MCP-1 and IL-6） in human umbilical vein endothelial cells （HUVECs）.Methods Human umbili-cal vein endothelial cells （HUVECs） were cultured and then stimulated with 50μg/ml ox-LDL for 24 hours. HUVECs were infected with adenovirus vectors over-expressing human KLF2 at an appropriate multiplicity of infection.KLF2-siRNA duplexes were transfected into HUVECs to silence the gene expression .HUVECs were collected at time points of 0 h, 3 h, 6 h, 12 h, 24 h and 48 h after infection.Real-time quantitative-PCR was performed to measure the expression of miR-146a at mRNA level.Silenced endogenous miR-146a using LNA-anti-miR-146 a was transfected into HUVECs with lipofectamine 2000 .The levels of MCP-1 and IL-6 in supernatants were detected by ELISA .Results KLF2 remarkably inhibited the expression of miR-146 a in unstimulated and ox-LDL-stimulated HUVECs in a time-dependent manner .The ox-LDL induced ex-pression of miR-146a, MCP-1 and IL-6 in HUVECs were significantly decreased by KLF2.Silenced expres-sion of miR-146a downr-egulated the ox -LDL induced expression of MCP-1 and IL-6 in HUVECs.Moreover, silenced miR-146 a could partly reverse the inhibitory effects of KLF 2 on ox-LDL induced expression of MCP-1 and IL-6 in HUVECs.Conclusion KLF2 inhibited ox-LDL induced expression of MCP-1 and IL-6 in HUVECs partly through down-regulating the expression of miR-146a. Key words: microRNA-146 a; Human umbilical vein endothelial cells; Oxidized low density lipoprotein; microRNA-146 a

Expression and clinical significance of plasma small RNA in patients with pancreatic cancer

The aim of this study was to identify six miRNAs expressed in plasma of patients with pancreatic cancer (PCa) and analyze their value as a diagnostic index of pancreatic cancer. Plasma total RNAs were extracted from 30 PCa patients and 26 normal controls, and the abundance of six microRNAs was measured using real-time PCR. The possibility to combine them with CA19-9 as diagnostic biomarkers was analyzed. The expression level of miR-21, miR-210, miR-155, miR-20a, miR-25 and miR-196a in plasma of patients with pancreatic cancer were 1.65×10(6), 5.98×10(4), 2.83×10(3), 3.47×10(6), 2.76×10(6), and 1.03×10(3) (copies/µl), while the normal controls were 4.08×10(5), 2.54×10(4), 8.55×10(2), 1.79×10(6), 9.32×10(5), and 4.67×10(2) (copies/µl), respectively, with a significant difference between the two groups (P < 0.05). The areas under the ROC curve of miR-21, miR-210, miR-155, miR-20a, miR-25 and miR-196a were 0.893, 0.810, 0.820, 0.766, 0.816 and 0.729, respectively. MiR-21 had the highest diagnostic value when it was used as diagnostic marker alone. The combination of miR-155 and miR-25 was more effective to distinguish PCa from normal than to be used alone, and the area under the ROC curve was 0.913 (95%CI 0.838-0.988) .When CA199 associated with miR -210 and miR-25, respectively, the areas under the ROC curves were 0.96 (95%CI was 0-1.0) and 0.942 (95% CI was 0.876-1.0), which were higher than CA199 alone (0.862, 95%CI was 0.748-0.975). There was a high improvement in diagnostic sensitivity and accuracy when miR-210 and miR-25 were combined with CA19-9, respectively. Plasma miR-21, miR-155, miR-25, miR-210 have diagnostic value for pancreatic cancer, and deserve further study.