Expression Levels Of miR-125b Research Articles

The effects of microRNA-181b on proliferation and invasion of pancreatic cancer cells

Objective To investigate the effects of microRNA-181b (miR-181b) on cell proliferation and invasion of pancreatic cancer cells. Methods MiR-181b expression levels in 3 pancreatic cancer cell lines PANC-1, BXPC-3, ASPC-1 and normal pancreatic HTERT-HPNE cells were observed by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). PANC-1 cells were transfected with 50 nmol/L miR-181b inhibitor or negative control using Lipofectamine 2000. After transfection, the expression of miR-181b was measured by RT-qPCR. Cell proliferation was tested by methylthiazol tetrazolium (MTT) assay. Cell invasion was detected by transwell. Bioinformatics prediction and luciferase activity tests were to verify phosphate and tension homology deleted on chromsome ten (PTEN) was the targeted gene of miR-181b. The expression levels of PTEN, matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) were measured by RT-qPCR. Results Compared with the HTERT-HPNE cells (1.00±0.06), the expression level of miR-181b in PANC-1 (2.84±0.16), BXPC-3 (1.74±0.18), ASPC-1 (2.21±0.20) was significantly increased. After transfection with 50 nmol/L miR-181b inhibitor, there was no obvious change in PANC-1 cell proliferation (P>0.05), and cell invasive ability was inhibited (P<0.01). Bioinformatics predicted PTEN might be the targeted gene of miR-181b. Luciferase experiments showed that luciferase activity of wild-type PTEN group fell by 39% after transfection of miR-181b mimics, but there was no statistical difference in the mutant group. After transfection with miR-181b inhibitor, PTEN was significantly increased (1.85±0.06 vs. 1.01±0.08, P<0.05), and MMP-2 (0.59±0.02 vs. 1.01±0.04) and MMP-9 (0.66±0.02 vs. 0.99±0.07) were significantly decreased (P<0.05). Conclusion These results demonstrate that miR-181b promotes proliferation and invasion in pancreatic cancer cells, which may be partly related to effecting MMP-2 and MMP-9 expression by PTEN. Therefore, it may be a new target for the biologic therapy for pancreatic cancer. Key words: Pancreatic cancer; MicroRNA-181b; Proliferation; Invasion

The effect of decitabine on the expression and methylation of the PPP1CA, BTG2, and PTEN in association with changes in miR-125b, miR-17, and miR-181b in NALM6 cell line.

Precursor B-cell acute lymphoblastic leukemia (B-ALL) is the most prevalent pediatric cancer. DNA methylation and changes in the microRNAs (miRNAs) expression are known to be important causes of B-ALL. Decitabine as a DNA methyltransferase inhibitor agent is able to induce hypomethylation in several tumor suppressor genes. Much evidence has proven BTG2, PPP1CA, and PTEN act as tumor suppressor genes in many malignancies. In this case control study, the messenger RNA (mRNA) expression of PPP1CA, BTG2, and PTEN genes using quantitative real-time polymerase chain reaction (rRT-PCR) in Nalm6 cell line and five patients suffer from ALL with mean age 5.6 years were determined in compare with seven normal healthy donors age and sex matched. qRT-PCR analysis revealed that the expression levels of PPP1CA, BTG2, and PTEN genes were significantly decreased in Nalm6 ([FC] = 0.46, [FC] = 0.046, [FC] = 0.54) and according to the Methylation-specific PCR (MSP) analysis, these genes were hypermethylated in Nalm6. In next step, the effects of decitabine treatment on the methylation and expression of these genes in association with changes in miR-125b, miR-17, and miR-181b expression levels were evaluated in optimal concentration 2.5 µM of decitabine. Our data showed that decitabine is able to restore the expression levels of aforementioned genes and downregulate expression levels of oncomiRs; including miR-125b, miR-17, and miR-181b in Nalm6 cell line. Therefore, it seems that decitabine can be used as a potential drug for the first line treatment of patients with B-ALL, but further in vivo investigation is necessary.

Matrine suppresses lipopolysaccharide-induced fibrosis in human peritoneal mesothelial cells by inhibiting the epithelial-mesenchymal transition.

Background:Peritoneal fibrosis is the primary reason that patients with end-stage renal disease (ESRD) have to cease peritoneal dialysis. Peritonitis caused by Gram-negative bacteria such as Escherichia coli (E. coli) were on the rise. We had previously shown that matrine inhibited the formation of biofilm by E. coli. However, the role of matrine on the epithelial-mesenchymal transition (EMT) in peritoneal mesothelial cells under chronic inflammatory conditions is still unknown.Methods:We cultured human peritoneal mesothelial cells (HPMCs) with lipopolysaccharide (LPS) to induce an environment that mimicked peritonitis and investigated whether matrine could inhibit LPS-induced EMT in these cells. In addition, we investigated the change in expression levels of the miR-29b and miR-129-5p.Results:We found that 10 μg/ml of LPS induced EMT in HPMCs. Matrine inhibited LPS-induced EMT in HPMCs in a dose-dependent manner. We observed that treatment with matrine increased the expression of E-cadherin (F = 50.993, P < 0.01), and decreased the expression of alpha-smooth muscle actin (F = 32.913, P < 0.01). Furthermore, we found that LPS reduced the expression levels of miR-29b and miR-129-5P in HPMCs, while matrine promoted the expression levels of miR-29b and miR-129-5P.Conclusions:Matrine could inhibit LPS-induced EMT in HPMCs and reverse LPS inhibited expressions of miR-29 b and miR-129-5P in HPMCs, ultimately reduce peritoneal fibrosis. These findings provide a potential theoretical basis for using matrine in the prevention and treatment of peritoneal fibrosis.

Circulating microRNA-133, microRNA-17 and microRNA-25 in serum and its potential diagnostic value in gastric cancer.

Gastric cancer is one of the most common malignancies in the world and is considered as the most lethal gastrointestinal cancer. microRNAs (miRNAs) can be very important in detecting a disease at an early stage. The aim of this study was to investigate the microRNA-17 (miR-17), miR-25, and miR-133b in the serum of gastric cancer subjects. Serum samples were obtained from 120 gastric cancers and 102 healthy subjects. We evaluated expression levels of miR-17, miR-25 and miR-133b by quantitative real-time polymerase chain reaction. Our results showed that in the patients with gastric cancer, the expression level of miR-17 and miR-25 were significantly increased compared with the control group (P < 0.5), while the expression level of miR-133b was significantly decreased in patient groups compared with control cases (P < 0.5). It seems that expression of miRNAs in Iranian patients with gastric cancer is similar to other patients in other populations. These findings suggested that miR-17, miR-25 and miR-133b could be introduced as potential diagnostic candidates for the detection in gastric cancer patients in the early stage.

Expression of miR-26b in ovarian carcinoma tissues and its correlation with clinicopathology.

The expression of microRNA (miR)-26b in ovarian carcinoma tissues, its correlation with clinicopathology, and its effect on diagnostic value and prognosis of ovarian cancer was investigated. A total of 74 patients with ovarian cancer (the study group) and 30 patients with benign ovarian tumors (the control group) in the Affiliated Hospital of Inner Mongolia Medical University from July 2011 to June 2013 were retrospectively analyzed. The expression of miR-26b in ovarian carcinoma tissues was detected by fluorescence reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and the correlation between the expression of miR-26b and the pathological features of ovarian carcinoma tissues and prognosis of patients was analyzed. The expression level of miR-26b in the study group (0.28±0.07) was significantly lower than that in the control group (0.54±0.11; P<0.050). There was no significant correlation between miR-26b expression and age, tumor type, exercise habit, smoking habit of patients with ovarian cancer (P>0.050), but there was close correlation between the miR-26b expression and lymph node metastasis, differentiation degree and pathological stage of patients with ovarian cancer (P<0.001). ROC curve showed that the area under curve (AUC) was 0.839, and when the maximum cut-off value was 0.815, the sensitivity and specificity of miR-26b in diagnosing the ovary was 84.932% and 77.936%, respectively. The 5-year overall survival rate in the low-expression group (61.54%) was significantly lower than that in the high-expression group (84.85; P=0.028). miR-26b is under-expressed in the ovary and has a close relationship with pathological stage, differentiation degree, and lymph node metastasis of ovarian cancer, which indicates that miR-26b is involved in the occurrence and development of ovarian cancer and is expected to be an effective indicator for treatment and diagnosis of ovarian cancer and the prognosis of patients.

Effect of miR-29b on rats with gestational diabetes mellitus by targeting PI3K/Akt signal.

To investigate the role of micro-ribonucleic acid-29b (miR-29b) in rats with gestational diabetes mellitus (GDM) through the phosphatidylinositol 3-kinase (PI3K)/serine/threonine kinase (Akt) signal and its mechanism by establishing rat models of GDM. Rat models of GDM were constructed, and then the expression levels of miR-29b, total PI3K, phosphorylated PI3K (p-PI3K), total Akt and phosphorylated Akt (p-Akt) in the model group and control group were measured via Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting assays, and the association between miR-29b expression and total PI3K expression was analyzed. In addition, miR-29b mimics and inhibitors were used to further explore the regulatory pathway, and the influences of miR-29b mimics and inhibitors on PI3K and Akt phosphorylation in GDM rats, characteristic indicators of oxidative stress such as superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in liver tissues of GDM rats, and fasting blood glucose in GDM rats were studied. Compared with those in the control group, miR-29b expression was lowered in rat models of GDM, while PI3K/Akt signal expression was increased. In rats with GDM, miR-29b expression was prominently negatively correlated with total PI3K expression (r=-0.777, p=0.007, p<0.01). MiR-29b mimics could reduce PI3K and Akt phosphorylation, increase SOD and CAT expression levels and decrease MDA content (p<0.05). Moreover, miR-29b mimics significantly lowered the blood glucose level in rats with GDM (p<0.05). MiR-29b mimics can alleviate oxidative stress and reduce blood glucose by inhibiting the PI3K/Akt signal transduction.

MiR-29b suppresses proliferation and mobility by targeting SOX12 and DNMT3b in pancreatic cancer.

Pancreatic cancer is one of the leading causes of solid carcinoma with the worst survival rate. The reasons for the worst survival rate include the lack of biomarkers for early detection, diagnosis at a late stage, and the limitation of the current therapy. Further study to investigate the underlying molecular mechanism in pancreatic cancer patients is necessary. A previous study showed that the miR-29b expression level is dysregulated, suggesting that it may serve an important function in pancreatic cancer. The CCK8 assay and the colony formation assay were used to detect the proliferation ability of the treated pancreatic cancer cells; a wound-healing assay and a transwell assay were used to test the migration and invasion ability and the interactive action of miR-29b and SOX12 or DNMT3b was examined by a luciferase assay. Cell proliferation, migration, and invasion were attenuated by miR-29b, whereas knockdown of SOX12 and DNMT3b could block SW1990 malignant activity. Further, the double luciferase assay showed that miR-29b can target SOX12 and DNMT3b directly by binding to their 3'-untranslated region. Finally, a rescue experiment was conducted by transfecting miR-29b and SOX12 overexpressed plasmid into cells. Cell proliferation, migration, and invasion inhibition induced by miR-29b were reversed by SOX12 overexpression, and revail of the expression of DNMT3b. MiR-29b suppressed proliferation, migration, and invasion by directly targeting SOX12 and DNMT3b in pancreatic cancer cells, and DNMT3b might be a target gene of SOX12.

MiR-29b Inhibits Ventricular Remodeling By Activating Notch Signaling Pathway in the Rat Myocardial Infarction Model.

To study the effect of miR-29b on myocardial infarction via Notch signaling pathway in rats. The rat acute myocardial infarction (AMI) models were established and were divided into AMI group, sham group and normal group (N = 10 in each group). HE (Hemotoxylin and eosin) staining was used to detect whether the model was constructed successfully. MiR-29b mimics, inhibitors, mimics negative control (NC) were transfected into H9c2 (2-1) cells. Then, cells were divided into a mimics group, inhibitor group, NC group, and blank group. The relative expression levels of miR-29b, Notch1, DII4 and Hesl were detected by qRT-PCR. The expression of NICD1 was detected by Western blotting. The rat AMI model was successfully constructed. Compared with normal and sham groups, the miR-29b expression was down-regulated, while the expression of Notch1, DII4 and Hesl was increased, and the NICD1 protein expression was increased in the myocardial infarction area of the AMI group (P < .05). Compared with the NC and blank groups, the relative expression of Notch1, DII4, Hesl and NICD1 were upregulated in the mimics group (P < .05), whereas the expression of Notch1, DII4, Hesl and NICD1 in the inhibitor group was decreased (P < .05). MiR-29b inhibited myocardial fibrosis and cardiac hypertrophy by activating the Notch signaling pathway and protected myocardium against myocardial infarction.

MicroRNA-26b acts as an antioncogene and prognostic factor in cervical cancer.

Cervical cancer is the second most frequent malignant neoplasm in women all over the world. MicroRNA-26b (miR-26b) has been reported to be downregulated and play a great role in many malignancies, nevertheless, there are scarce studies on cervical cancer. The purpose of the present study was to detect how miR-26b is involved in cervical carcinoma. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was utilized to detect the expression levels of miR-26b and Jagged1 (JAG1) mRNA. Transwell assay was applied to calculate the cell migration and invasion capacity. Luciferase reporter assay was employed to determine JAG1 as a target of miR-26b. The results revealed that miR-26b is downregulated in cervical cancer tissues and cells compared with paracancerous tissues and normal cervical epithelial cells. The low expression of miR-26b in cervical cancer demonstrated that miR-26b inhibits cell migration and invasion, as measured by Transwell assay. JAG1 was verified to be a target of miR-26b and have a negative correlation with miR-26b, as detected by luciferase reporter assay. In addition, miR-26b was found to suppress cell migration and invasion via mediating JAG1 expression, which impact is partially reversed by JAG1. In conclusion, miR-26b suppresses cell migration and invasion of cervical cancer through directly targeting JAG1. It is suggested that miR-26b/JAG1 axis may present a new target for the treatment of cervical cancer.

Decreased miR-29b expression is associated with airway inflammation in chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is a common chronic airway inflammatory disease. MicroRNAs are shown to be involved in the regulation of inflammation. We investigated the role of microRNA-29b (miR-29b) in the airway inflammation in COPD. The expression of miR-29b in the lung and plasma was examined. The target of miR-29b, bromodomain protein 4 (BRD4), was predicted by online algorithms and verified in human bronchial epithelial (HBE) cells. The expression of BRD4, interleukin (IL)-8, and IL-6 in the lung was also examined. The role of miR-29b in the inflammatory cytokine expression of airway epithelial cells was studied using an in vitro model system. In total, 60 subjects were recruited, including 10 nonsmokers, 24 smokers, and 26 patients with COPD. Both lung and plasma miR-29b are decreased in patients with COPD, and miR-29b expression levels are correlated with pulmonary function and inflammation. BRD4 is increased in the lung of patients with COPD and is correlated with miR-29b and IL-8 expression. miR-29b regulates cigarette smoke extract (CSE)-induced IL-8 expression by targeting BRD4 in HBE cells. The antioxidant N-acetylcysteine prevents CSE-induced miR-29b downregulation and BRD4 and IL-8 upregulation. Our findings indicate that miR-29b may participate in the airway inflammation in COPD by regulating inflammatory cytokine expression through targeting BRD4, plasma miR-29b may serve as a biomarker for disease severity in COPD, and oxidative stress may contribute to the decrease of miR-29b induced by cigarette smoke.

Correlation Between miR-125b Expression and Liver Fibrosis in Patients with Chronic Hepatitis C

Objectives: The study aimed to investigate the role of miR-125b as a non-invasive biomarker in chronic hepatitis C. Methods: An observational study was conducted on 94 treatment-naive HCV-infected patients (mean age 49.8 ± 11.5 years, 59.6% females). Liver fibrosis was assessed by transient elastography (TE) and the expression of miR-125b in plasma was quantified by real-time PCR. Results: All patients were infected with HCV genotype 1b and had active viral replication, 42.6% had significant cytolysis, and 73.4% had increased serum gamma-glutamyl transferase (GGT) values. Significant fibrosis (liver stiffness measured by TE of > 7.1 kPa) was present in 61.7% of the patients. No significant associations were found between miR-125b expression and baseline HCV viral load (P = 0.56), IL28B polymorphisms (P = 0.5), alpha-fetoprotein levels (P = 0.27), and patients’ gender (P = 0.13) or age (P = 0.5). In a univariate analysis, the miR-125b expression level was significantly correlated with ALT (P = 0.001) and GGT levels (P < 0.0001). An up-regulated expression of miR-125b was found in plasma samples from patients with advanced liver fibrosis as compared to those with mild/moderate fibrosis [mean miR-125b value = 0.002 versus 0.001 (P = 0.02)]. In a multiple regression analysis, an upregulated miR-125b expression level remained independently in association only with significant fibrosis and increased GGT level (P = 0.026; R2 = 0.242). Conclusions: An up-regulated miR-125b expression might be an indicator of severe liver fibrosis in patients with chronic hepatitis C, independent of the viral replication level.

Aging-Related Changes in Cognition and Cortical Integrity are Associated With Serum Expression of Candidate MicroRNAs for Alzheimer Disease.

Evidence has shown that microRNAs (miRNAs) are involved in molecular pathways responsible for aging and prevalent aging-related chronic diseases. However, the lack of research linking circulating levels of miRNAs to changes in the aging brain hampers clinical translation. Here, we have investigated if serum expression of brain-enriched miRNAs that have been proposed as potential biomarkers in Alzheimer's disease (AD) (miR-9, miR-29b, miR-34a, miR-125b, and miR-146a) are also associated with cognitive functioning and changes of the cerebral cortex in normal elderly subjects. Results revealed that candidate miRNAs were linked to changes in cortical thickness (miR-9, miR-29b, miR-34a, and miR-125b), cortical glucose metabolism (miR-29b, miR-125b, and miR-146a), and cognitive performance (miR-9, miR-34a, and miR-125b). While both miR-29b and miR-125b were related to aging-related structural and metabolic cortical changes, only expression levels of miR-125b were associated with patterns of glucose consumption shown by cortical regions that correlated with executive function. Together, these findings suggest that serum expression of AD-related miRNAs are biologically meaningful in aging and may play a role as biomarkers of cerebral vulnerability in late life.

Correlation between estrogen plasma level and miRNAs in muscle of Piedmontese cattle

A loss-of-function mutation of the myostatin gene has a very high prevalence in the Piedmontese cattle breed. The effect of such mutation is a double-muscle phenotype because of hypertrophy. However, differences in muscle mass development can still be detected in individuals of this breed. Such differences must be generated by other factors controlling skeletal muscle development. MicroRNAs are short noncoding RNA molecules that modulate gene expression at a post-transcriptional level. MicroRNAs have been demonstrated to be involved in skeletal muscle development, and some of them are controlled by steroid hormone signaling. Data on estrogen signaling are lacking, whereas more studies have been carried out on the effect of androgens. We aimed at identifying putative estrogen responsive miRNAs that might be involved in skeletal muscle development. At a slaughterhouse, we collected muscle samples from longissimus dorsi and blood samples. Blood 17β-estradiol concentration was assessed, and RNA was extracted from muscle samples. The animals we sampled were divided into groups according to estrogen blood concentration, and through qPCR expression, levels of 7 muscle-related miRNAs were evaluated. We found that miR-26b (P < 0.01), miR-27a-5p (P < 0.05), miR-27b (P < 0.05), and miR-199a-3p (P < 0.01) were differentially expressed among experimental groups. Expression levels of miR-26b were reduced approximately 50% in samples with a low blood estrogen concentrations, and the other miRNAs showed a tendency to increase their expression levels when blood estrogen levels were higher. The variations of the circulating concentrations of estrogen in Piedmontese cattle might influence muscle mass development through miRNAs and thus contribute to individual variability in a breed with a high prevalence of a myostatin point mutation.

Down-Regulation of MiR-125b Reverses Drug Resistance of Doxorubicin-Resistant Leukemia Cells

To investigate whether the down-regulation of miR-125b can reverse the drug-resistence of doxorubicine-resistant leukemia cell lines or not, so as to explore a new method for treatment of drug-resistant leukemia patients. The expression levels of miR125b in doxorubicine drug-sensitive and doxorubicine drug-resistant leukemia cell lines.HL-60, K562 and HL-60/Dox, the K562/Dox were detected by using RT-qPCR; the up-regulation or inhibition of miR-1256 expression in HL-60/Dox were performed by electroporation transfection, then the viability of cells treated with doxorubicine of different concentration was detected by CCK-8 method, the proliferation inhibition curve of cells was drawed, and the IC50 was calculated. The miR-125b expression was obviously up-regulated in drug-resistant cell lines HL-60/DOX and K562/DOX, as compared with HL-60 and K562 cell lines. The miR-125b expression level in HL-60/DOX and K562/DOX cells was 15 times and 5 times higher than that in HL-60 and K562 cells, respectively. The up-regulating or inhibiting expression of miR-125b in HL-60/DOX cells found that the proliferation inhibition rate in cells transfected with miR-125b mimic significantly decreased, compared with control group (P<0.01), while the proliferation inhibition rate in cells transfected with miR-125b inhibitor significantly increased, compared with control group(P<0.01). The miR-125b expression in HL-60/Dox and K562/Dox cells has been up-regulated, down-regulation of miR-125b expression can reverse the drug resistance of leukemia cells to doxorubicine.

Steroid hormone calcitriol and its analog tacalcitol inhibit miR-125b expression in a human breast cancer MCF-7 cell line

MiR-125b belongs to the class of microRNAs, which are short endogenous non-coding RNAs that negatively regulate gene expression at the post-transcriptional level. Recently, it was reported that miR-125b was found to promote migration and invasion of MCF-7 cells and was involved in chemotherapeutic resistance. Decreasing miR-125b expression would have potential therapeutic significance in preventing dissemination of breast cancer cells. The objective of this study was to evaluate miR-125b expression levels in MCF-7 cells following treatment with 1,25-dihydroxyvitamin D3 (calcitriol) and 1,24-dihydroxyvitamin D3 (tacalcitol), active metabolite and synthetic analog of vitamin D3, respectively. We found that treatment with both calcitriol and tacalcitol caused a decrease in miR-125b expression. In addition, treatment with calcitriol and tacalcitol resulted in an increase in the level of pro-apoptotic BAK1 protein encoded by the target gene of miR-125b. We are discussing the putative mechanism of inhibition of the miR-125b expression by vitamin D receptor (VDR) agonists and we suggest that calcitriol and tacalcitol may be used as a miR-125b inhibitor in breast cancer cells expressing VDR.

The deregulation of miR-133b is associated with poor prognosis in bladder cancer

BackgroundMicroRNAs (miRNAs) are single-stranded, endogenous, non-coding RNAs that are increased or decreased in almost all cancer types, and they paly crucial roles in the tumorigenesis as well as development. Materials and methods90 patients diagnosed with bladder cancer were enrolled in the present study. The bladder cancer tissues or adjacent normal tissues were obtained from the tumor area or adjacent normal zone. The expression level of miR-133b was examined by quantitative real-time polymerase chain reaction assay (qRT-PCR). Survival curves were displayed by the Kaplan-Meier method, and differences between two survival curves were calculated by the log-rank test. ResultsThe expression levels of miR-133b in bladder tissues were significantly decreased when compared with the matched adjacent normal bladder tissues (P < 0.05). Moreover, miR-133b expression levels are significantly associated with lymphatic invasion (P = 0.026), distant metastasis (P = 0.025), tumor grade (P = 0.038), as well as the muscle invasion status (P < 0.001). The log-rank test indicated that patients with decreased miR-133b expression underwent poorer overall survival (P = 0.007). Furthermore, multivariate Cox regression analysis showed that the expression level of miR-133b (P = 0.024) was an independent factor for predicting the overall survival in patients with bladder cancer. ConclusionsThe present study showed that miR-133b might be associated with bladder cancer progression, and its down-regulation might be a biomarker for poor prognosis of bladder cancer.

MiR-10b regulates the proliferation and apoptosis of pediatric acute myeloid leukemia through targeting HOXD10.

To investigate the role of miR-10b in the proliferation and apoptosis of acute myeloid leukemia (AML), and to explore the underlying mechanism. The expression level of miR-10b in clinical AML cases and cell lines was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The interaction between miR-10b and homeobox D10 (HOXD10) was confirmed by qRT-PCR, Western blotting and Luciferase assay. The effect of miR-10b on biological functions of AML cell line (HL60) was analyzed in vitro. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and colony formation assay were used to detect the proliferation and colony formation ability of AML cells, respectively. Meanwhile, flow cytometry and TUNEL staining were applied to measure cell cycle and apoptosis of AML cells, respectively. miR-10b was significantly up-regulated in AML cases and cell lines. The potential target genes of miR-10b were analyzed by three public databases. Results showed that HOXD10 was a direct target of miR-10b. QRT-PCR, Western blotting and luciferase assay confirmed the regulatory effect of miR-10b on HOXD10. Overexpression of miR-10b accelerated the proliferation and colony formation ability of AML cells. Meanwhile, miR-10b overexpression decreased the percentage of AML cells in the G0/G1 phase when compared with S phase, and suppressed the apoptosis of AML cells. However, the addition of HOXD10 could reverse the effects of miR-10b. MiR-10b could regulate the proliferation, colony formation, cell cycle and apoptosis of AML cells through targeting HOXD10, indicating that miR-10b might be a potential therapeutic target for the treatment of AML.

Retraction: MiR-125b Increases Nasopharyngeal Carcinoma Radioresistance By Targeting A20/NF-κB Signaling Pathway.

Radioresistance poses a major challenge in nasopharyngeal carcinoma (NPC) treatment, but little is known about how miRNA regulates this phenomenon. In this study, we investigated the function and mechanism of miR-125b in NPC radioresistance, one of upregulated miRNAs in the radioresistant NPC cells identified by our previous microarray analysis. We observed that miR-125b was frequently upregulated in the radioresistant NPCs, and its increment was significantly correlated with NPC radioresistance, and was an independent predictor for poor patient survival. In vitro radioresponse assays showed that miR-125b inhibitor decreased, whereas miR-125b mimic increased NPC cell radioresistance. In a mouse model, therapeutic administration of miR-125b antagomir dramatically sensitized NPC xenografts to irradiation. Mechanistically, we confirmed that A20 was a direct target of miR-125b and found that miR-125b regulated NPC cell radioresponse by targeting A20/NF-κB signaling. With a combination of loss-of-function and gain-of-function approaches, we further showed that A20 overexpression decreased while A20 knockdown increased NPC cell radioresistance both in vitro and in vivo Moreover, A20 was significantly downregulated while p-p65 (RelA) significantly upregulated in the radioresistant NPCs relative to radiosensitive NPCs, and miR-125b expression level was negatively associated with A20 expression level, whereas positively associated with p-p65 (RelA) level. Our data demonstrate that miR-125b and A20 are critical regulators of NPC radioresponse, and high miR-125b expression enhances NPC radioresistance through targeting A20 and then activating the NF-κB signaling pathway, highlighting the therapeutic potential of the miR-125b/A20/NF-κB axis in clinical NPC radiosensitization. Mol Cancer Ther; 16(10); 2094-106. ©2017 AACR.

Expression of microRNA-181b in sepsis-induced acute lung injury rats and its correlation with the levels of inflammatory cytokines

Objective To investigate the differential expression of microRNA (miRNA, miR)-181b in sepsis-induced acute lung injury (ALI) and its correlation with interleukin (IL)-6 and tumor necrosis factor (TNF)-α. Methods Twenty-four SD male rats were randomly divided into 4 groups: normal control group, 3-h sepsis group, 6-h sepsis group, and 12-h sepsis group in equal number. Sepsis-induced ALI model was induced by intraperitoneal injection of lipopolysaccharide (LPS, 10 mg/kg). The injury of lung tissue was evaluated histopathologically. The expression of miR-181b was detected by reverse transcription polymerase chain reaction (RT-PCR), and IL-6 and TNF-α were measured by enzyme linked immunosorbent assay (ELISA) and the correlation analysis was carried out. Results The lung tissue hematoxylin -eosin staining indicated ALI model was successfully established in the sepsis group. The expression level of miR-181b in the lung tissue of the sepsis group (6 h) was 0.41±0.05 respectively, which was significantly lower than that in the normal control group (P=0.002). The expression of miR-181b in the lung tissue of the sepsis group (6 h) was negatively correlated with the level of IL-6 and TNF-α (r=-0.985, P=0.000; r=-0.990, P=0.000). Conclusion The expression level of miR-181b was significantly down-regulated in sepsis-induced ALI, and had a significantly negative correlation with IL-6 and TNF-α. Key words: MicroRNA-181b; Sepsis; Acute lung injury; Inflammatory cytokines

Expression of p53 and its effect on microRNA-29b expression in the process of normal human conjunctival fibroblasts transforming to myofibroblast

Objective To investigate the relationship between p53 and miR-29b in myofibroblasts (MFs) induced from human Tenon fibroblasts (HTFs). Methods Human Tenon tissues were obtained from strabismus surgery patients in the First Affiliated Hospital of Anhui Medical University from March to July 2016.The primary HTFs were cultured by tissue culture method and passaged to 2-4 generations.The cells were divided into 4 groups: HTFs group cells were normally cultured, the TGF-β1 activation group cells was activated with 10 ng/ml tumor necrosis factorβ1(TGF-β1), the siRNA transfection group and the negative control group were transfected with p53 siRNA and negative siRNA, respectively.The expression of p53 protein and cell migration ability of HTFs and MFs were detected by immunohistochemical and cell scratch assay.The relative expression level of miR-29b and p53 mRNA in each group was detected by real-time PCR.This study followed the Helsinki declaration and the patients or their guardians signed informed consent. Results The primary HTFs had long fusiform shape, and were arranged in a spiral shape.After activated by TGF-β1, the cells were transformed into MFs, and the cell morphology was irregular and the arrangement was disordered.The staining intensity of p53 protein in TGF-β1 activation group was significantly stronger than that in HTFs group(t=-10.384, P<0.05). The migration distance of TGF-β1 activation group cells was significantly larger than that of HTFs group (P<0.05). The relative expression level of p53 mRNA was 1.00±0.00, 3.95±2.61, 0.06±0.06 and 0.98±0.16 in HTFs group, TGF-β1 activation group, siRNA transfection group and negative control group, respectively; the relative expression levels of miR-29b was 1.00±0.00, 0.54±0.09, 5.10±2.31 and 1.03±0.09 in HTFs group, TGF-β1 activation group, siRNA transfection group and negative control group, respectively; the overall differences were statistically significant (p53 mRNA: F=6.688, P=0.006; miR-29b: F=13.640, P=0.000). The relative expression of p53 mRNA was significantly higher and the relative expression of miR-29b was significantly lower in the TGFβ1 activation groups than those in the HTFs group and siRNA transfection group, the differences were statistically significant (all at P<0.05). Conclusions During the process of HTFs transforming to MFs, the expression of p53 is upregulated and the expression of miR-29b is downregulated, Supressing the expression of p53 can upregulate the expression of miR-29b.There may exist interactions between p53 and miR-29b, which regulate the formation of fibrotic scars. Key words: Glaucoma; Scar; p53; MicroRNA-29b