Expression Levels Of miR-125b Research Articles

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

MicroRNA-10b expression predicts long-term survival in patients with solid tumor.

Numerous studies have evaluated the significance of the microRNA-10b (miR-10b) in the development and progression of many cancers. Their findings revealed that increased expression of miR-10b is associated with unfavorable prognosis in patients with cancer. A total of 1,834 patients from 19 studies were included in this study. A significantly shorter overall survival was observed in patients with increased expression of miR-10b (hazard ratio [HR] = 1.99, 95% confidence interval [CI]: 1.51-2.61). Statistical significance was also observed in subgroup meta-analysis stratified by the cancer type, cutoff value, analysis type, and sample size. Also, patients with a high expression level of miR-10b had a poorer disease-free survival rate (HR = 1.18, 95% CI: 1.05-1.33). In addition, the pooled odds ratios (ORs) showed that increased miR-10b was also associated with positive lymph node metastasis (OR = 2.09, 95% CI: 1.45-3.03), distant metastasis (OR = 2.40, 95% CI: 1.57-3.67), tumor size (OR = 3.86, 95% CI: 2.25-6.64), and poor clinical stage (OR = 5.02, 95% CI: 3.37-7.47). A systematic literature search was conducted on a number of electronic databases, including PubMed, Embase, Web of Science, China National Knowledge Infrastructure, Springer, Google Scholar, and Gene expression omnibus. We retrieved the relevant articles to examine the association between the miR-10b expression levels and patients' prognosis. The meta-analysis was conducted using the RevMan 5.2 software and Stata SE12.0 software. High miR-10b expression was correlated with poor clinical outcome, which indicated the potential clinical use of miR-10b as a molecular biomarker for cancer, particularly in assessing prognosis for patients with cancers. Further studies should be performed to verify the clinical utility of miR-10b in human solid tumors.

OncomiR-10b hijacks the small molecule inhibitor linifanib in human cancers

The pervasive role of microRNAs (miRNAs) in cancer pathobiology drives the introduction of new drug development approaches such as miRNA inhibition. In order to advance miRNA-therapeutics, meticulous screening strategies addressing specific tumor targets are needed. Small molecule inhibitors represent an attractive goal for these strategies. In this study, we devised a strategy to screen for small molecule inhibitors that specifically inhibit, directly or indirectly, miR-10b (SMIRs) which is overexpressed in metastatic tumors. We found that the multi-tyrosine kinase inhibitor linifanib could significantly inhibit miR-10b and reverse its oncogenic function in breast cancer and liver cancer both in vitro and in vivo. In addition, we showed that the efficacy of linifanib to inhibit tyrosine kinases was reduced by high miR-10b levels. When the level of miR-10b is high, it can “hijack” the linifanib and reduce its kinase inhibitory effects in cancer resulting in reduced anti-tumor efficacy. In conclusion, our study describes an effective strategy to screen for small molecule inhibitors of miRNAs. We further propose that miR-10b expression levels, due to the newly described “hijacking” effect, may be used as a biomarker to select patients for linifanib treatment.

Expression and Function of miR-181b in Diffuse Large B Cell Lymphoma

To investigate the expression and function of miR-181b in diffuse large B cell lymphoma (DLBCL). The lymphoid tissues of 124 patients with DLBCL examined and treated in our hospital from March 2010 to March 2012 were used as the DLBCL group. The healthy lympaid dissases in another 64 patients with non lymphaoma were selected as the control group. The expression levels of miR-181b in two groups were detected, and the expression levels of miR-181b in lymphoid tissues of DLBCL patients with different clinical features were compared. The survival of DLBCL patients with different levels of miR-181b expression was compared. The relative expression level of miR-181b in lymph tissues of DLBCL patients was significantly higher than that in healthy lymphoid tissues (P<0.05). The higher the Ann Arbor staging was, the higher the relative expression of miR-181b was in lymphoid tissues (P<0.05), and the higher the IPI score was, the higher the relative expression of miR-181b was in lymphoid tissues (P<0.05). The 5-year survival rate of the patients with miR-181b low expression was significantly higher than that of patients with miR-181b high expression (P<0.05). There are differences in the expression of miR-181b in lymphoid tissues of DLBCL patients with different clinical stages and prognosis. It may become an effective indicator for the diagnosis and prognosis evaluation of DLBCL.

MicroRNA-26b suppresses tumorigenicity and promotes apoptosis in small cell lung cancer cells by targeting myeloid cell leukemia 1 protein

The aim of this study was to investigate the role of microRNA-26b (miR-26b) in regulating the proliferation, migration, and apoptosis of small cell lung cancer (SCLC) cells. First, we examined the expression level of miR-26b in human normal fetal lung fibroblasts (NFLFs) and three SCLC cell lines NCI-H466, NCI-H1688, and NCI-H196. In the following experiments, the three SCLC cell lines were transfected with miR-26b mimic and inhibitor. Cell growth and survival, as well as migration and invasion capacities were determined by MTT, colony formation, Transwell migration and invasion, and wound healing assays. Cell apoptosis, production of reactive oxygen species, and mitochondrial membrane potential were also measured in the three cell lines following various treatments. As a result, we found that the level of miR-26b was significantly lower in SCLC cells than in NFLFs. Additionally, transfection with miR-26b mimic could inhibit proliferation, colony formation, and migration, as well as induce apoptosis in these SCLC cell lines; while miR-26b inhibitor showed the opposite effects. Further mechanistic experiment revealed that miR-26b could suppress the expression of myeloid cell leukemia 1 protein (Mcl-1) and the 3′-untranslated region (3′-UTR) of Mcl-1 may be the direct binding site of miR-26b, suggesting that the effect of miR-26b may be mediated by targeting Mcl-1. Collectively, our findings offer a new insight into the role of miR-26b in the pathogenesis of SCLC, and provide primary evidence supporting the potential of miR-26b-based therapy for the treatment of SCLC.

PO-343 The microRNA-146b-IRAK1 signalling axis in tumour recurrence of papillary thyroid cancer

IntroductionPapillary thyroid cancer (PTC) is the most common tumour subtype of thyroid cancer. However, not all PTCs are responsive to current surgical and radioiodine treatment. The well-established clinical prognostic factors include tumour size, lymph node/distal metastasis, and extrathyroidal invasion. The RET/PTC-RAS-BRAF linear molecular signalling cascade is known to mediate PTC pathogenesis. However, whether presence of BRAF mutation, the most common genetic alteration in PTC, can affect PTC behaviour and prognosis is controversial. MicroRNAs (miRNAs) have been labelled as promising molecular prognostic markers in several tumour types. However, the underlying mechanism of miR-146b in relation to its oncogenic behaviour in PTC and its putative targets remain to be fully elucidated.Material and methodsWe used genome-wide microarray, computational analysis and 3 UTR reporter gene assays to identify IRAK1 as a miR-146b target gene. In vitro gain/loss-of-function experiments were fur- ther performed to determine the effects of IRAK1 on proliferation, colony formation, and wound- healing in PTC cancer cell lines. Expression levels of miR-146b and IRAK1 of 50 cases of PTC and its adjacent normal thyroid specimens were assessed via quantitative real-time polymerase chain reaction.Results and discussionsMicroarray expression profile revealed that the mRNA level of IRAK1 gene was down- regulated by miR-146b. The 3 UTR of IRAK1 mRNA was found to be a molecular target of miR-146b posttranscriptional repression in BCPAP cells by reporter gene assays. MiR-146b promoted the migration and proliferation of PTC cells by downregulating IRAK1 expression, whereas restoration of IRAK1 expression reversed this effect. In addition, the expression of IRAK1 mRNA was significantly lower in PTC clinical tissue samples than normal adjacent thyroid specimens and showed a strong inverse correlation with the expression of miR-146b in PTC specimens. Our results demonstrated that IRAK1 is a direct target of miR-146b and has functional roles to inhibit various aggressive PTC cell activities.ConclusionIn conjunction with current therapeutic regimens, targeting the miR-146b-IRAK1 axis not only may provide a potential approach for PTC management but also serve as a unique supplemental tool for diagnosis and predicting prognosis for PTC in the near future.

Regulation of tamoxifen resistance in estrogen receptor positive MCF-7 breast cancer cells by microRNA-10b targeting histone deacetylase 4

Objective To investigate the effect of microRNA-10b (miR-10b) targeting histone deacetylase 4 (HDAC4) on the resistance of estrogen receptor positive (ER+ ) MCF-7 breast cancer cells to tamoxifen. Methods ER+ and tamoxifen resistant MCF-7 cell line MCF-7-TR was established, and miR-10b expression level in MCF-7 cells and MCF-7-TR cells was detected by reverse transcription real-time quantitative polymerase chain reaction (Real-time PCR). Methyl thiazol tetrazolium (MTT) and trypan blue were used to detect the proliferation and activity of MCF-7 cells and MCF-7-TR cells transfected with pre-miR-10b, miR-10b inhibitor and siHDAC4 at different tamoxifen concentrations (0, 5, 10, 15, 20, 30 μmol/L), respectively. The expression of HDAC4 protein was detected by Western blotting. Results Taken expression level and invasive ability of miR-10b in MCF-7 breast cancer cells as the standard, the miR-10b expression level and invasion ability of MCF-7-TR cell breast cancer cells were 7.2±1.1 and 5.3±1.3 respectively. With the increase of the concentration of tamoxifen, decreased the rate of cell proliferation in MCF-7 cells, which is most obvious, followed by MCF-7-TR+ miR-10b inhibitor + siHDAC4 and MCF-7+ pre-miR-10b+ HDAC4 cells, and the proliferation of MCF-7-TR+ siHDAC4 cells and MCF-7-TR cells decreased the rate of the lowest. The migration ability of MCF-7 cells and MCF-7-TR+ miR-10b cells of the inhibitor decreased most, MCF-7+ pre-miR-10b+ and tamoxifen (10 μmol/L) MCF-7-TR+ cells and tamoxifen (10 μmol/L) had no significant difference in the ability of cell migration (t=1.116, P=0.272), but was significantly higher than that of MCF-7 cells and MCF-7-TR+ miR-10b cells (t=0.124, 10.769, 14.569, 11.577, P=0.000). The expression level of HDAC4 protein in MCF-7+ pre-miR-10b cells was lower than that in MCF-7 cells, MCF-7-TR cells, HDAC4 protein expression level, MCF-7-TR+ miR-10b inhibitor cells, MCF-7+ siHDAC4 cells HDAC4 protein expression level was lower than MCF-7 cells. Conclusion The miR-10b-HDAC4 signaling pathway may be a molecular mechanism of tamoxifen resistance in breast cancer. Key words: Breast cancer; Estrogen receptor positive; Tamoxifen resistance; MicroRNA-10b; Histone deacetylase 4

Clinicopathological significance of miR-26, miR-107, miR-125b, and miR-203 in head and neck carcinomas.

MicroRNAs play a role in the development and progression of head and neck squamous cell carcinomas (HNSCC). Our aim was to study the expression of miR-26, miR-107, miR-125b, and miR-203 in primary HNSCC with and without lymph node metastasis and their clinicopathological significance. The expression of microRNAs in primary HNSCC with lymph node metastasis (n=16) and their matched lymph node, as well as primary tumors without metastasis (n=16), were determined by quantitative RT-PCR and analyzed with clinicopathological features and survival. The expression levels of miR-26 (p<.05) and miR-125b (p<.01) were higher in metastatic primary HNSCC, while levels of miR-203 (p<.01) were lower. The expression of the microRNAs was associated with clinicopathological features, including miR-26 high expression and N stage (p=.04), poor differentiation (p=.005) and recurrence (p=.007), miR-125b high expression and N stage (p=.0005) and death (p=.02), and low levels of miR-203 and N stage (p=.04). The high expression of miR-26 was associated with shortened disease-free survival, and high miR-125b expression was an independent risk factor for poor disease-specific survival. These findings suggest that miR-26 and miR-125b may be associated with the progression and metastasis of HNSCC and that miR-203 is associated with a more favorable prognosis.

Tumor Expression of miR-10b, miR-21, miR-143 and miR-145 Is Related to Clinicopathological Features of Gastric Cancer in a Central European Population.

In Western countries, most patients with gastric cancer (GC) present in advanced stages. Therefore, there is imminent clinical need for novel diagnostic and prognostic biomarkers. Deregulation of microRNAs has been reported as a frequent event in GC development in a number of studies. Our study validated the potential of microRNAs to serve as diagnostic and prognostic biomarkers in patients with GC from the Central European population. Using quantitative real-time polymerase chain reaction, expression levels of six microRNAs (miR-10b, -21, -93, -107, - 143, and -145) were examined in 67 tumor tissues and 67 paired adjacent gastric tissues, and correlated with clinicopathological features of GC patients. Expression levels of miR-10b, miR-21, miR-93, and miR-107 were significantly higher in GC samples compared to non-tumor tissue. Furthermore, the expression levels of miR-10b, miR-143, and miR-145 positively correlated with advanced stages, and increased expression of miR-10b, miR-21 and miR-145 was significantly associated with worse prognosis of gastric cancer patients. Our results indicate that selected tissue microRNAs have the potential to serve as relevant diagnostic and prognostic biomarkers of GC in a central European population.

Higher expression of miR-133b is associated with better efficacy of erlotinib as the second or third line in non-small cell lung cancer patients.

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (gefitinib, erlotinib and afatinib) are indicated as first-line therapy in patients with non-small cell lung cancer (NSCLC) whose tumors harbor activating mutations in the EGFR gene. Erlotinib is also used in second and third-line therapy for patients whose tumors have wild type EGFR but to date there are no validated biomarkers useful to identify which patients may benefit from this treatment. The expression level of four miRNAs: miR-133b, -146a, -7 and -21 which target EGFR was investigated by real-time PCR in tumor specimens from NSCLC patients treated with erlotinib administered as the second or third line. We found that miR-133b expression level better discriminated responder from non-responder patients to erlotinib. Higher levels of miR-133b in NSCLCs were associated with longer progression-free survival time of patients. Functional analyses on miR-133b through transfection of a miR-133b mimic in A549 and H1299 NSCLC cell lines indicated that increasing miR-133b expression level led to a decreased cell growth and altered morphology but did not affect sensitivity to erlotinib. The detection of miR-133b expression levels in tumors help in the identification of NSCLC patients with a better prognosis and who are likely to benefit from second and third-line therapy with erlotinib.

PD46-12 SETD2-MEDIATED LOSS OF MIR-10B DETERMINES MAP4K4 ACTIVATION IN RENAL CELL CARCINOMA

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology I1 Apr 2018PD46-12 SETD2-MEDIATED LOSS OF MIR-10B DETERMINES MAP4K4 ACTIVATION IN RENAL CELL CARCINOMA libin Yan, heng Li, yucong Zhang, hua Xu, and zhangqun Ye libin Yanlibin Yan More articles by this author , heng Liheng Li More articles by this author , yucong Zhangyucong Zhang More articles by this author , hua Xuhua Xu More articles by this author , and zhangqun Yezhangqun Ye More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.2160AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To investigate the potential function and mechanism of SETD2/H3K36me3-miRNA-10b-MAPKs pathway in renal cell carcinoma(RCC). METHODS The expression level of miR-10b was evaluated through the meta-analysis of eligible studies reported miR-10b expression in multiple cancers and miRNA probe test in RCC tissue. The function of miR-10b and its down-stream molecular MAPKs was investigated in RCC cell lines via Transwell, MTS and EdU assays. The mechanism of SETD2-miR-10b-MAPKs was predicted by TCGA and Targetscan database. After knocking down SETD2 or miR-10b, this pathway was verified by chromatin immunoprecipitation(ChIP), luciferase assays et al. In the xenograft assay, three groups of Balb/c nude mices were treated with shSETD2, shSETD2+mimics-miR-10b and sh-LacZ in 786-O cell line, respectively. RESULTS SETD2 and miR-10b were downregulated in RCC. After up regulation of miR-10b by RNAi interference, the ability of proliferation, invasion and migration of RCC cells was suppressed. In addition, the result showed that the expression level of miR-10b was down-regulated after knock-down of SETD2. miR-10b can suppress the expression of MAP4K4,MAP3K7 and CDK6 directly and further influence phosphorylation of JNK. In xenograft assay, there was a dramatic decrease in tumor volume and weight in the shSETD2+mimics-miR-10b and sh-LacZ group compared with sh-SETD2 group. CONCLUSIONS miR-10b was the direct target of SETD2 and miR-10b can inhibit the expression of MAP4K4, MAP3K2 and MAP3K7, thus affecting the phosphorylation of JNK protein in MAPK signaling pathway and playing the role of tumor suppression; Targeting the SETD2/H3K36me3-miRNA-10b-MAPKs pathway might be an effective molecular targeted therapy to renal clear cell carcinoma. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e895-e896 Advertisement Copyright & Permissions© 2018MetricsAuthor Information libin Yan More articles by this author heng Li More articles by this author yucong Zhang More articles by this author hua Xu More articles by this author zhangqun Ye More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Involvement of methylation of MicroRNA-122, \u2212125b and -106b in regulation of Cyclin G1, CAT-1 and STAT3 target genes in isoniazid-induced liver injury

BackgroundThis investigation aimed to evaluate the role of methylation in the regulation of microRNA (miR)-122, miR-125b and miR-106b gene expression and the expression of their target genes during isoniazid (INH)-induced liver injury.MethodsRats were given INH 50 mg kg− 1·d− 1 once per day for 3, 7, 10, 14, 21 and 28 days and were sacrificed. Samples of blood and liver were obtained.ResultsWe analysed the methylation and expression levels of miR-122, miR-125b and miR-106b and their potential gene targets in livers. Liver tissue pathologies, histological scores and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities changed, indicating the occurrence of liver injury. Relative expression levels of miR-122, miR-125b and miR-106b genes in the liver decreased after INH administration and correlated with the scores of liver pathology and serum AST and ALT activities, suggesting that miR-122, miR-125b and miR-106b are associated with INH-induced liver injury. The amount of methylated miR-122, miR-125b and miR-106b in the liver increased after INH administration and correlated with their expression levels, suggesting the role of methylation in regulating miRNA gene expression. Two miR-122 gene targets, cell cycle protein G1 (Cyclin G1) and cationic amino acid transporter-1 (CAT-1), also increased at the mRNA and protein levels, which suggests that lower levels of miR-122 contribute to the upregulation of Cyclin G1 and CAT-1 and might play a role in INH-induced liver injury. Signal transducer and activator of transcription 3 (STAT3) was a common target gene of miR-125b and miR-106b, and its expression levels of mRNA and protein increased after INH administration. The protein expression of phosphorylated (p)-STAT3 and the mRNA expression of RAR-related orphan receptor gamma (RORγt) regulated by p-STAT3 also increased. Meanwhile, the mRNA and protein expression of interleukin (IL)-17 regulated by RORγt, and the mRNA and protein expression of CXCL1 and MIP-2 regulated by IL-17 increased after INH administration. These results demonstrate that lower levels of hepatic miR-125b and miR-106b contribute to the upregulation of STAT3 in stimulating the secretion of inflammatory factors during INH-induced liver injury.ConclusionsOur results suggested that DNA methylation probably regulates the expression of miRNA genes (miR-122, miR-125b, and miR-106b), affecting the expression of their gene targets (Cyclin G1, CAT-1, and STAT3) and participating in the process of INH-induced liver injury.

MicroRNA-10b expression in breast cancer and its clinical association.

MicroRNAs (miRNAs) are short non-coding RNA molecules that play a significant role in many types of cancers including breast cancer. In the current study, we evaluated the expression levels of microR-10b (miR-10b) in 115 breast cancer patients from Sichuan Cancer Center. Real time reverse transcription-PCR was used to assess miR-10b expression. Clinical data including disease stage, survival status, age, ER/PR/HER2 status, molecular subtypes, tumor size, lymph node status and Ki-67 expression levels were correlated with miR-10b expression levels. Our data showed that the miR-10b expression is correlated with disease stage, living status and tumor sizes. We also found that miR-10b expression levels are higher in the lymph node positive group and the Ki-67 higher scoring group (score > 20). No statistically significant differences were observed based on age or molecular sub-type grouping. In conclusion, miR-10b may be a biomarker for breast cancer and is a potential treatment target.

RETRACTED ARTICLE: Effects of Panax notoginseng saponins on severe acute pancreatitis through the regulation of mTOR/Akt and caspase-3 signaling pathway by upregulating miR-181b expression in rats

BackgroundIn China, Panax notoginseng has been used to treat oxidative stress-related diseases for a long time. Panax notoginseng saponins is an extract from Panax notoginseng Ledeb. Its therapeutic potential is related to antioxidant activity, but related mechanisms are still unclear. The study aims to assess the protection effects of Panax notoginseng saponins in the taurocholate-induced rat model of acute pancreatitis (AP) and explore underlying mechanisms.MethodsA rat model of severe acute pancreatitis (SAP) was established in rats induced with taurocholate. Panax notoginseng saponins was firstly administered in the treatment group via intravenous injection. After 2 h, taurocholate administration was performed. After 24 h, the expression levels of miR-181b, Beclin1, LC3-II, Akt and mTOR from pancreas tissues were measured by Western Blotting and RT-PCR. Then the expression levels of Caspase-3 and Blc-2 were determined by immunohistochemistry. Apoptosis was assessed by the TUNEL assay. Amylase and lipase in serum were determined by ELISA and pancreatic water contents in pancreatic tissue were measured. After eosin and hematoxylin staining, the histologic analysis was performed.ResultsAfter SAP induction by taurocholate and the treatment with Panax notoginseng saponins for 24 h, we detected the up-regulated miR-181b, the reduced Bcl-2 expression, the increased activity of mTOR/Akt, the blocked Beclin1 and LC3-II expressions, and the enhanced Caspase-3 expression. Serum lipase and amylase levels were significantly decreased in the treatment group of Panax notoginseng saponins compared to the control group. Histological analysis results verified the attenuation effects of Panax notoginseng saponins on taurocholate-induced pancreas injury, apoptosis, and autophagy.ConclusionBy up-regulating the miR-181b expression level, Panax notoginseng saponins significantly reduced taurocholate-induced pancreas injury and autophagy and increased apoptosis. The significant protection effects of Panax notoginseng saponins suggested its potential in treating taurocholate induced-acute pancreatitis.

Overexpression of MicroRNA-29b Decreases Expression of DNA Methyltransferases and Improves Quality of the Blastocysts Derived from Somatic Cell Nuclear Transfer in Cattle.

MicroRNA (miR)-29b plays a crucial role during somatic cell reprogramming. The aim of the current study was to explore the effects of miR-29b on the developmental competence of bovine somatic cell nuclear transfer (SCNT) embryos, as well as the underlying mechanisms of action. The expression level of miR-29b was lower in bovine SCNT embryos at the pronuclear, 8-cell, and blastocyst stages compared with in vitro fertilized embryos. In addition, miR-29b regulates the expression of DNA methyltransferases (Dnmt3a/3b and Dnmt1) in bovine SCNT embryos. We further investigated SCNT embryo developmental competence and found that miR-29b overexpression during bovine SCNT embryonic development does not improve developmental potency and downregulation inhibits developmental potency. Nevertheless, the quality of bovine SCNT embryos at the blastocyst stage improved significantly. The expression of pluripotency factors and cellular proliferation were significantly higher in blastocysts from the miR-29b overexpression group than the control and downregulation groups. In addition, outgrowth potential in blastocysts after miR-29b overexpression was also significantly greater in the miR-29b overexpression group than in the control and downregulation groups. Taken together, these results demonstrated that miR-29b plays an important role in bovine SCNT embryo development.

MicroRNA-125b Affects Vascular Smooth Muscle Cell Function by Targeting Serum Response Factor

Background/Aims: Increasing evidence links microRNAs to the pathogenesis of peripheral vascular disease. We recently found microRNA-125b (miR-125b) to be one of the most significantly down‑regulated microRNAs in human arteries with arteriosclerosis obliterans (ASO) of the lower extremities. However, its function in the process of ASO remains unclear. This study aimed to investigate the expression, regulatory mechanisms, and functions of miR-125b in the process of ASO. Methods: Using the tissue explants adherent method, vascular smooth muscle cells (VSMCs) were prepared for this study. A rat carotid artery balloon injury model was constructed to simulate the development of vascular neointima, and a lentiviral transduction system was used to overexpress serum response factor (SRF) or miR-125b. Quantitative real‑time PCR (qRT‑PCR) was used to detect the expression levels of miR‑125b and SRF mRNA. Western blotting was performed to determine the expression levels of SRF and Ki67. In situ hybridization analysis was used to analyze the location and expression levels of miR-125b. CCK-8 and EdU assays were used to assess cell proliferation, and transwell and wound closure assays were performed to measure cell migration. Flow cytometry was used to evaluate cell apoptosis, and a dual-luciferase reporter assay was conducted to examine the effects of miR‑125b on SRF. Immunohistochemistry and immunofluorescence analyses were performed to analyze the location and expression levels of SRF and Ki67. Results: miR-125b expression was decreased in ASO arteries and platelet-derived growth factor (PDGF)-BB-stimulated VSMCs. miR-125b suppressed VSMC proliferation and migration but promoted VSMC apoptosis. SRF was determined to be a direct target of miR-125b. Exogenous miR-125b expression modulated SRF expression and inhibited vascular neointimal formation in balloon-injured rat carotid arteries. Conclusions: These findings demonstrate a specific role of the miR-125b/SRF pathway in regulating VSMC function and suggest that modulating miR-125b levels might be a novel approach for treating ASO.

32 MicroRNA-29b Improves the Quality and Developmental Potential of Blastocysts Derived from Somatic Cell Nuclear Transfer in Cattle

MicroRNAs (miRNAs) are small non-coding RNAs with important roles in diverse cellular processes. miR-29b plays a crucial role during somatic cell reprogramming. However, studies of the function of miR-29b in embryogenesis are limited. The aim of the current study was to explore the effects of miR-29b on the developmental competence of bovine somatic cell nuclear transfer (SCNT) embryos as well as the underlying mechanisms of action. The expression level of miR-29b was lower in bovine SCNT embryos at the pronuclear, 8-cell, and blastocyst stages compared with IVF embryos (P &lt; 0.05). To determine the function of miR-29b in the bovine SCNT embryo, we microinjected a miR-29b mimic and inhibitor into bovine SCNT zygotes. The results showed that miR-29b significantly decreased the expression of Dnmts (Dnmt3a/3b and Dnmt1) in bovine SCNT embryos (P &lt; 0.05). We further investigated SCNT embryo developmental competence and found that miR-29b overexpression during bovine SCNT embryonic development does not improve developmental potency (P &gt; 0.05) but down-regulation inhibits developmental potency (P &lt; 0.05). Although miR-29b overexpression does not improve the developmental potency of bovine SCNT embryos, the quality of bovine SCNT embryos at the blastocyst stage improved significantly (P &lt; 0.05). The expression of pluripotency factors (OCT4 and SOX2) and cellular proliferation rate were significantly higher in blastocysts from the miR-29b overexpression group than the control and down-regulation groups (P &lt; 0.05). In addition, outgrowth potential in blastocysts after miR-29b overexpression was also significantly greater in the miR-29b overexpression group than in the control and down-regulation groups (P &lt; 0.05). Taken together, these results demonstrated that miR-29b plays an important role in bovine SCNT embryo development.

MiR-125b regulates the drug-resistance of breast cancer cells to doxorubicin by targeting HAX-1.

MircroRNAs (miRNAs) are considered as essential regulators in the tumorigenesis and chemoresistance of multiple cancer types. In the present study, it was demonstrated that the expression levels of miR-125b were significantly downregulated in the tissues of patients with breast cancer (BC), as well as the BC cell lines in vitro. To study the association between chemoresistance and miR-125b in BC, doxorubicin (DOX)-resistant MCF-7 (MCF-7/R) cells were established, and gain- and loss-of-function experiments were performed. It was demonstrated that the overexpression of miR-125b increased the sensitivity of MCF-7/R cells to DOX. Furthermore, it was revealed that the sensitization of miR-125b mimics to DOX-induced cell death was regulated by the hematopoietic cell-specific protein 1-associated protein X-1 (HAX-1) vector and HAX-1 small interfering RNA. These results emphasized the notable function of miR-125b and its target of HAX-1 in regulating DOX-resistance. In addition, it was demonstrated that the miR-125b mimics promoted the loss of the mitochondrial membrane potential and the generation of reactive oxygen species induced by DOX treatment in MCF-7/R cells. These data suggest that the miR-125b-HAX-1-mitochondria pathway has a notable function in the treatment of DOX-resistant BC cells, which may provide a novel target for the chemotherapy of BC.

Restoration of miR-29b exerts anti-cancer effects on glioblastoma

BackgroundGlioblastoma multiforme (GBM) is known as one of the most fatal forms of cancer. MicroRNAs have been widely implicated in the regulation of mammalian development and pathogenesis. The brain-enriched miR-29 subfamilies are known to be exclusively expressed in the developing brain, and they are aberrantly down-regulated in GBM. This study aims to elucidate the role of miR-29b in GBM development and the feasibility of therapeutic targeting using conjugated nanoparticles.MethodsAfter confirmation of miR-29b expression levels in GBM tissues by analysis of open source data, the anticancer effect of miR-29b was tested by the introduction of syn-hsa-miR-29b-3p in the A172 GBM cell line. In vitro studies of cell viability and apoptosis and ex vivo study using GBM tissue slice cultures from 3 patients and nanoparticle delivery of miR-29b were performed.ResultsWe discovered an increase in apoptotic cell populations with the introduction of miR-29b in the GBM cell line. An established human-derived GBM tissue slice culture system confirmed the anticancer effect of miR-29b-conjugated nanoparticles. Using PCR array, we found that exogenous miR-29b inhibits the expression of COL1A2, COL3A1, COL4A1, ELN, ITGA11, MMP24, and SPARC, which mediates an anticancer effect.ConclusionsmiR-29b may serve as a putative therapeutic molecule when its expression is restored using a nanoparticle delivery system in GBM.

MiR-133b induces chemoresistance of osteosarcoma cells to cisplatin treatment by promoting cell death, migration and invasion.

As an important chemotherapeutic agent for the treatment of osteosarcoma, the effectiveness of cisplatin is considered to be due to its unique properties, which allow it to penetrate the cell membrane and form various DNA-platinum adducts, resulting in genetic alterations or DNA damage. However, chemoresistance to cisplatin remains a major challenge for its use and chemotherapeutic effects. In the present study, an isogenic model of a cisplatin resistant osteosarcoma cell line, MG63-DDP, was generated from the original MG63 cell line. The expression level of microRNA (miR)-133b in the MG63-DDP cisplatin-resistant osteosarcoma cell line was analyzed by reverse transcription-quantitative polymerase chain reaction (PCR). Cisplatin-DNA adduct formation, cell death (carboxyfluorescein succinimidyl ester/propidium iodide staining) and clonogenic survival assays (crystal violet staining) were performed, comparing various cell types. The effect of miR-133b on migration (scratch wound assay) and invasion (Transwell assay) was also evaluated. Characterization studies have previously revealed an increased level of miR-133b in MG63-DDP cells compared with normal MG63 cells. Upregulation of miR-133b was associated with the accumulation of cisplatin-DNA adducts and an increase in cisplatin-induced cell death. Furthermore, increased miR-133b expression levels enhanced the migration and invasion of MG63 cells under cisplatin stress. Concordantly, in MG63-DDP cells the neutralization of miR-133b demonstrated opposite effects, as compared with the upregulation of miR-133b. To the best of our knowledge, the present study demonstrated for the first time that cisplatin-resistant MG63 cells exhibit an increased level of miR-133b expression. The endogenous expression level of miR-133b is sufficient for inducing cisplatin resistance, which suggests that miR-133b may be a biomarker for cisplatin resistance in osteosarcoma.