Expression Of miR-200a Research Articles

DNA methylation and miRNA expression in colon adenomas compared with matched normal colon mucosa and carcinomas.

Dysregulation of DNA methylation patterns and non-coding RNA, including miRNAs, has been implicated in colon cancer, and these changes may occur early in the development of carcinoma. In this study, the role of epigenetics as early changes in colon tumorigenesis was examined through paired sample analysis of patient-matched normal, adenoma and carcinoma samples. Global methylation was assessed by genomic 5-methyl cytosine (5-mC) and long interspersed nuclear element-1 (LINE-1) promoter methylation by pyrosequencing. KRAS mutations were also assessed by pyrosequencing. Expression of miRNA, specifically, two microRNA genes-miR-200a and let-7c-was analysed using RT-qPCR. Differences in global methylation in adenomas were not observed, compared with normal tissue. However, LINE-1 methylation was decreased in adenomas (p=.056) and carcinomas (p=.011) compared with normal tissue. Expressions of miRNA, miR-200a and let-7c were significantly higher in adenomas than normal tissues (p=.008 and p=.045 respectively). Thus the significant changes in LINE-1 methylation and microRNA expression in precancerous lesions support an early role for epigenetic changes in the carcinogenic process. Epigenetic characteristics in adenomas may provide potential diagnostic and prognostic therapeutic targets early in cancer development at the adenoma stage.

Differential Expression of Serum TUG1, LINC00657, miR-9, and miR-106a in Diabetic Patients With and Without Ischemic Stroke.

Background: Ischemic stroke is one of the serious complications of diabetes. Non-coding RNAs are established as promising biomarkers for diabetes and its complications. The present research investigated the expression profiles of serum TUG1, LINC00657, miR-9, and miR-106a in diabetic patients with and without stroke. Methods: A total of 75 diabetic patients without stroke, 77 patients with stroke, and 71 healthy controls were recruited in the current study. The serum expression levels of TUG1, LINC00657, miR-9, and miR-106a were assessed using quantitative real-time polymerase chain reaction assays. Results: We observed significant high expression levels of LINC00657 and miR-9 in the serum of diabetic patients without stroke compared to control participants. At the same time, we found marked increases of serum TUG1, LINC00657, and miR-9 and a marked decrease of serum miR-106a in diabetic patients who had stroke relative to those without stroke. Also, we revealed positive correlations between each of TUG1, LINC00657, and miR-9 and the National Institutes of Health Stroke Scale (NIHSS). However, there was a negative correlation between miR-106a and NIHSS. Finally, we demonstrated a negative correlation between LINC00657 and miR-106a in diabetic patients with stroke. Conclusion: Serum non-coding RNAs, TUG1, LINC00657, miR-9, and miR-106a displayed potential as novel molecular biomarkers for diabetes complicated with stroke, suggesting that they might be new therapeutic targets for the treatment of diabetic patients with stroke.

Research on Mechanism of miR-106a Nanoparticles Carrying Dexmedetomidine in Regulating Recovery and Metabolism of Nerve Cells in Hypoxia-Reoxygenation Injury.

We studied the mechanism of miR-106a nanoparticles carrying dexmedetomidine (DEX) in regulating the recovery and metabolism of nerve cells in hypoxia-reoxygenation injury. Hippocampus neuron model in hypoxia-reoxygenation injury was prepared in vitro. Study groups were randomly divided into control set, ischemic reperfusion (IR) set, dexmedetomidine (DEX) set, miR-106a-nanoparticles (NPs) set and set of dexmedetomidine (DEX) and miR-106a-NPs. We studied miR-106a expression, proliferative and apoptotic activity, secretion of IL-6 and tumor necrosis factor (TNF)-α, quantity of Phosphocreatine (PCr), adenosine triphosphate (ATP) and total adenine nucleotide, and also content of reactive oxygen species (ROS) and superoxide dismutases (SOD). Expressions of of Bax, Bcl-2 and NF-κB were also detected. Results showed that the expression of miR-106a in hippocampus neuron was reduced, while proliferation was reduced and apoptotic activity was increased. The secretions of IL-6 and TNF-α were increased, while the quantities of Phosphocreatine (PCr), adenosine triphosphate (ATP) and total adenine nucleotide were reduced. Bax expression was also increased and Bcl-2 expression was reduced. Moreover, ROS content was increased and SOD activity was reduced, while the NF-κB presentation was increased. The above-mentioned changes could be reversed in IR set, DEX set and miR-106a-NPs set. The action was more notable in the DEX and miR-106a-NPs sets. Finally, the proliferation in hippocampus neuron in hypoxia-reoxygenation injury could be prompted and apoptosis could be restrained by DEX and miR-106a-NPs. The secretion of inflammatory factors could be restrained through restraining the inflammatory pathway and oxidative stress. The energy metabolism could therefore be improved effectively and recovery of nerve cells in HBI could be improved.

Up-regulation of miR-20a ameliorates sevoflurane anesthesia-induced cognitive impairment in rats by targeting EphA4


 
 
 
 Purpose: To evaluate the role of miR-20a in sevoflurane (SEV)-induced cognitive impairment in rats and to elucidate the mechanism of action.
 Methods: A SEV-induced cognitive impairment rat model was developed. The Morris water maze test and fear assay were carried out to assess impaired learning and memory. A cellular SEV-impaired model was developed and the miR-20a level was measured in the animal and cellular models. TUNEL staining and immunoblot assay were performed to determine the SEV effect on apoptosis. Bioinformatic analysis and luciferase assay were conducted to identify the target of miR-20a action. A rescue assay involving miR-20a overexpression in cellular and animal models was developed and used to evaluate function of miR-20a in cognitive defects.
 Results: The rats showed significant cognitive impairment upon SEV treatment, which inhibited the expression of miR-20a and promoted neuronal apoptosis. Further findings identified EphA4 as a target of miR-20a, which regulates its expression. Overexpression of miR-20a in rats effectively reduced cognitive dysfunction and apoptosis of hippocampus somatic cells caused by SEV treatment.
 Conclusion: Evidently, miR-20a ameliorates SEV anesthesia-induced cognitive impairment in rats and thus has the potential to serve as a therapeutic target for the treatment of post-operative cognitive dysfunction.
 
 
 

Regulation of retinal amacrine cell generation by miR-216b and Foxn3.

The mammalian retina contains a complex mixture of different types of neurons. We find that microRNA miR-216b is preferentially expressed in postmitotic retinal amacrine cells in the mouse retina, and expression of miR-216a/b and miR-217 in retina depend in part on Ptf1a, a transcription factor required for amacrine cell differentiation. Surprisingly, ectopic expression of miR-216b directed the formation of additional amacrine cells and reduced bipolar neurons in the developing retina. We identify the Foxn3 mRNA as a retinal target of miR-216b by Argonaute PAR-CLIP and reporter analysis. Inhibition of Foxn3, a transcription factor, in the postnatal developing retina by RNAi increased the formation of amacrine cells and reduced bipolar cell formation. Foxn3 disruption by CRISPR in embryonic retinal explants also increased amacrine cell formation, whereas Foxn3 overexpression inhibited amacrine cell formation prior to Ptf1a expression. Co-expression of Foxn3 partially reversed the effects of ectopic miR-216b on retinal cell formation. Our results identify Foxn3 as a novel regulator of interneuron formation in the developing retina and suggest that miR-216b likely regulates Foxn3 and other genes in amacrine cells.

Regular Intake of Green Tea Polyphenols Suppresses the Development of Nonmelanoma Skin Cancer through miR-29-Mediated Epigenetic Modifications.

Previously, we and others have shown that the regular intake of green tea polyphenols (GTPs) reduces ultraviolet B (UVB) radiation-induced skin cancer by targeting multiple signaling pathways, including DNA damage, DNA repair, immunosuppression, and inflammation. Here, we determine the effect of GTPs on UVB-induced epigenetic changes, emphasizing DNA hypermethylation in UV-exposed skin and tumors and their association with miR-29, a key regulator of DNA methyltransferases (DNMTs). Skin cancer was induced in SKH-1 hairless mice following repeated exposures of UVB radiation (180 mJ/cm2, three times/week, 24 weeks) with or without GTPs supplementation (0.2%) in drinking water. Regular intake of GTPs inhibited tumor growth by hindering the cascade of DNA hypermethylation events. GTPs supplementation significantly blocked UVB-induced DNA hypermethylation in the skin (up to 35%; p < 0.0001) and in tumors (up to 50%; p < 0.0001). Experimental results showed that the levels of DNA hypermethylation were higher in GTPs-treated mice than in the control group. The expressions of miR-29a, miR-29b, and miR-29c were markedly decreased in UV-induced skin tumors, and GTPs administration blocked UVB-induced miR-29s depletion. Furthermore, these observations were verified using the in vitro approach in human skin cancer cells (A431) followed by treatment with GTPs or mimics of miR-29c. Increased levels of miR-29 were observed in GTPs-treated A431 cells, resulting in increased TET activity and decreased DNA hypermethylation. In conclusion, UVB-mediated miR-29 depletion promotes DNA hypermethylation and leads to enhanced tumor growth by silencing tumor suppressors. Regular intake of GTPs rescued UVB-induced miR-29 depletion and prevented tumor growth by maintaining reduced DNA hypermethylation and activating tumor suppressors. Our observations suggest that miR-based strategies and regular consumption of GTPs could minimize the risk of UVB-induced skin cancers and contribute to better management of NMSCs.

Dynamic MicroRNA Expression Profiles During Embryonic Development Provide Novel Insights Into Cardiac Sinus Venosus/Inflow Tract Differentiation.

MicroRNAs have been explored in different organisms and are involved as molecular switches modulating cellular specification and differentiation during the embryonic development, including the cardiovascular system. In this study, we analyze the expression profiles of different microRNAs during early cardiac development. By using whole mount in situ hybridization in developing chick embryos, with microRNA-specific LNA probes, we carried out a detailed study of miR-23b, miR-130a, miR-106a, and miR-100 expression during early stages of embryogenesis (HH3 to HH17). We also correlated those findings with putative microRNA target genes by means of mirWalk and TargetScan analyses. Our results demonstrate a dynamic expression pattern in cardiac precursor cells from the primitive streak to the cardiac looping stages for miR-23b, miR-130a, and miR-106a. Additionally, miR-100 is later detectable during cardiac looping stages (HH15-17). Interestingly, the sinus venosus/inflow tract was shown to be the most representative cardiac area for the convergent expression of the four microRNAs. Through in silico analysis we revealed that distinct Hox family members are predicted to be targeted by the above microRNAs. We also identified expression of several Hox genes in the sinus venosus at stages HH11 and HH15. In addition, by means of gain-of-function experiments both in cardiomyoblasts and sinus venosus explants, we demonstrated the modulation of the different Hox clusters, Hoxa, Hoxb, Hoxc, and Hoxd genes, by these microRNAs. Furthermore, we correlated the negative modulation of several Hox genes, such as Hoxa3, Hoxa4, Hoxa5, Hoxc6, or Hoxd4. Finally, we demonstrated through a dual luciferase assay that Hoxa1 is targeted by miR-130a and Hoxa4 is targeted by both miR-23b and miR-106a, supporting a possible role of these microRNAs in Hox gene modulation during differentiation and compartmentalization of the posterior structures of the developing venous pole of the heart.

Diagnostic and Prognostic Value of miRNAs in Hepatoblastoma: A Systematic Review With Meta-Analysis.

Background and aim: Increasing evidence has revealed the valuable diagnostic and prognostic applications of dysregulated microRNAs (miRNAs) in hepatoblastoma (HB), the most common hepatic malignancy during childhood. However, these results are inconsistent and remain to be elucidated. In the present study, we aimed to systematically compile up-to-date information regarding the clinical value of miRNAs in HB. Methods: Articles concerning the diagnostic and prognostic value of single miRNAs for HB were searched from databases. The sensitivity (SEN), specificity (SPE), positive and negative likelihood ratios (PLR and NLR), diagnostic odds ratio (DOR), area under the curve (AUC), and hazard ratios (HRs) were separately pooled to explore the diagnostic and prognostic performance of miRNA. Subgroup and meta-regression analyses were further carried out only in the event of heterogeneity. Results: In all, 20 studies, involving 264 HB patients and 206 healthy individuals, met the inclusion criteria in the 6 included literature articles. For the diagnostic analysis of miRNAs in HB, the pooled SEN and SPE were 0.76 (95% CI: 0.72-0.80) and 0.75 (95% CI: 0.70-0.80), respectively. Moreover, the pooled PLR was 2.79 (95% CI: 2.12-3.66), NLR was 0.34 (95% CI: 0.26-0.45), DOR was 10.24 (95% CI: 6.55-16.00), and AUC was 0.83, indicating that miRNAs had moderate diagnostic value in HB. For the prognostic analysis of miRNAs in HB, the abnormal expressions of miR-21, miR-34a, miR-34b, miR-34c, miR-492, miR-193, miR-222, and miR-224 in patients were confirmed to be associated with a worse prognosis. The pooled HR was 1.74 (95% CI: 1.20-2.29) for overall survival and 1.74 (95% CI: 1.31-2.18) for event-free survival, suggesting its potential as a prognostic indicator for HB. Conclusion: To the best of our knowledge, this is the first comprehensive systematic review and meta-analysis that examines the diagnostic and prognostic role of dysregulated miRNAs in HB patients. The combined meta-analysis results supported the previous individual finds that miRNAs might provide a new, noninvasive method for the diagnostic and prognostic analyses of HB.

Expression of miR-571 and miR-20a in Breast Cancer Patients as Diagnostic Biomarkers

Expression of miR-571 and miR-20a in Breast Cancer Patients as Diagnostic Biomarkers

Integration of exosomal miR-106a and mesothelial cells facilitates gastric cancer peritoneal dissemination

BackgroundMetastatic organotropism is considered the end stage of malignancy with the mechanism still have some mysteries that have not been disclosed. Although the role of miR-106a is well studied, its involvement in the formation of peritoneal metastasis transported by exosomes is less discussed. MethodsGastric cancer (GC)-derived exosomes were identified by transmission electron microscopy and the integration with peritoneal mesothelial cells (PMC) was confirmed by PKH-26 staining. Cell phenotype was assessed by EdU and flow cytometry. MiR-106a and its targets were authenticated by luciferase reporter assay. The mesothelial-to-mesenchymal transition (MMT) and extracellular matrix (ECM) degeneration were determined and morphological transformation was measured by transwell assay. Immunodeficient mouse was conducted to investigate the tumorigenesis and tumor growth. The final was tissue analysis. ResultsMiR-106a enrichment in GC-exosomes can be delivered and integrated into PMC to establish a proper pre-metastatic niche (PMN). We found that PMC could internalize exosomal-miR-106a and lead to increased apoptotic rate and decreased proliferative vitality. The direct target Smad7 and TIMP2 were proved to be effectively reduced by translocation of exosomal miR-106a. We confirmed that exosomal miR-106a was able to induce MMT and accelerate ECM through targeting Smad7 and TIMP2 to activate TGF-β pathway. Animal model investigated that the tumorigenesis and tumor growth could be influenced by exosomes, and exosomal-miR-106a could promote the formation of xenograft tumor. Tissue analysis verified the ectopic miR-106a expression in gastric cancer metastasis. ConclusionOur data suggest that exosomal miR-106a facilitates gastric cancer peritoneal dissemination by integrating PMC to destroy mesothelial barrier.

LncRNA cardiac autophagy inhibitory factor is downregulated in rheumatoid arthritis and suppresses the apoptosis of fibroblast-like synoviocytes by promoting the maturation of miRNA-20a.

In this study, we aimed to investigate the effects of LncRNA cardiac autophagy inhibitory factor (CAIF) and miR-20a on the apoptosis of synovial cells in rheumatoid arthritis (RA) and the regulatory mechanism. Between May 2018 and March 2020, a total of 62 RA patients (24 males, 38 females; mean age: 55.2±4.9 years; range, 42 to 68 years) and 62 controls (24 males, 38 females; mean age: 55.3±4.8 years; range, 41 to 68 years) were included in this study. Plasma samples were collected from all participants. The expression levels of CAIF, mature miR-20a, and miR-20a precursor in these plasma samples were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Correlations were analyzed using linear regression analysis. Overexpression of CAIF was achieved in human fibroblast-like synoviocytes (HFLSs) and the expression levels of mature miR-20a and miR-20a precursor were determined using RT-qPCR. Cell apoptosis was analyzed by cell apoptosis assay. The CAIF was downregulated in RA and positively correlated with the expression of mature miR-20a. In HFLSs, LPS treatment resulted in downregulation of both CAIF and miR-20a in a dose-dependent manner. In HFLSs, overexpression of CAIF did not affect the expression of miR-20a precursor, but upregulated the expression of mature miR-20a. Cell apoptosis analysis showed that overexpression of CAIF and miR-20a inhibited the apoptosis of HFLSs induced by LPS. The combination of overexpression of CAIF and miR-20a showed a stronger effect. The CAIF may suppress the apoptosis of HFLSs in RA by promoting the maturation of miR-20a.

Expression patterns of miR-34a, miR-125b, miR-221 and antioxidant gene NRF2 in plasma samples of patients with atherosclerosis

Atherosclerosis is one of the main reasons of cardiovascular diseases, the most common cause of death in the world. NRF2 is a critical transcriptional factor that regulates oxidative stress response and contributes to the pathogenesis of atherosclerosis. This study aims to compare the expression levels of miR-34a, miR-125b, miR- 221 and NRF2 in blood samples of patients with atherosclerosis and of controls to find a novel link between microRNAs, oxidative stress and atherosclerosis. miR-34a, miR-125b, miR-221 and NRF2 relative expressions were analysed in 26 atherosclerosis patients and 25 healthy controls by q-RT-PCR assay. The receiver operating characteristic curve (ROC) was used to assess the diagnostic values of miR-34a, miR-125b, miR-221 and NRF2 by comparing the area under the curves (AUC) to differentiate between atherosclerosis and control samples. miR-34a and NRF2 were significantly upregulated, whereas miR-125b and miR-221 were significantly downregulated in patients compared with healthy controls. The ROC curves suggested high diagnostic value of miR-221 (AUC: 0.7477), miR-125b (AUC: 0.8523) and NRF2 (AUC: 0.838) for detection of atherosclerosis. Our results suggest that circulating miR-34a, miR-125b and miR-221 levels can be used as novel biomarkers for detection of atherosclerosis by targeting NRF2.

Multi-Omics Reveal the Immunological Role and the Theragnostic Value of miR-216a/GDF15 Axis in Human Colon Adenocarcinoma.

Colon adenocarcinoma (COAD) is the most common type of gastrointestinal cancer and is still the third leading cause of cancer-related mortality worldwide. Accurate screening tools for early diagnosis and prediction of prognosis and precision treatment strategies are urgently required to accommodate the unmet medical needs of COAD management. We herein aimed to explore the significance of the microRNA (miR)-216a/growth differentiation factor 15 (GDF15) axis in terms of clinical value, tumor immunity, and potential mechanisms in COAD by using multi-omic analysis. The gene expression levels of miR-216a and GDF15 showed an increase in the COAD group compared to those of the normal group. The expression of miR-216a presented a negative correlation with GDF15 in COAD tumor tissue. The use of an in vitro luciferase reporter assay and bioinformatic prediction revealed that miR-216a-3p acted toward translational inhibition on GDF15 by targeting its 3′untranslated region (UTR) site. High miR-216a expression was associated with decreased overall survival (OS), while the high expression of GDF15 was associated with increased OS. Enriched type 1 T-helper (Th1), enriched regulatory T (Treg), enriched eosinophils, and decreased nature killer T-cells (NKTs) in COAD tumor tissue may play counteracting factors on the tumor-regulatory effects of miR-216a and GDF15. In addition, high GDF15 expression had associations with suppressed immunoinhibitory genes and negative correlations with the infiltration of macrophages and endothelial cells. The enrichment analysis revealed that GDF15 and its co-expression network may be implicated in mitochondrial organization, apoptosis signaling, and endoplasmic reticulum (ER) stress response. The Genomics of Drug Sensitivity in Cancer (GDSC) and Cancer Therapeutics Response Portal (CTRP) analysis identified that Gemcitabine acted as a precision treatment for COAD when GDF15 expression was low. This study supports the miR-216a/GDF15 axis as a diagnostic/prognostic panel for COAD, identifies Th1, Treg, eosinophils, and NKTs as counteracting factors, indicates potential relationships underlying immunomodulation, mitochondrial organization, apoptotic signaling, and ER stress and unveil Gemcitabine as a potential drug for the development of treatment strategy when combined with targeting GDF15.

MicroRNA-20a Suppresses Tumor Proliferation and Metastasis in Hepatocellular Carcinoma by Directly Targeting EZH1.

PurposeHepatocellular carcinoma (HCC), a worldwide leading cause of morbidity and mortality, is the most frequent primary liver tumor. Most HCC patients are diagnosed with advanced liver cancer, resulting in a very low 5-year survival rate. Thus, there is an urgent need for the development of targeted therapies. In this study, we aimed to investigate the effect and mechanism of the miR-20a/EZH1 axis on the proliferation and metastasis of HCC and the inhibitory effect of the EZH1/EZH2 inhibitor UNC1999 on HCC.Materials and MethodsThe expression of miR-20a in human HCC tissues and cell lines was detected using quantitative real-time PCR (qRT-PCR). The expressions of proteins were analyzed with immunohistochemistry and Western blotting. Luciferase assay was used to verify whether miR-20a targets EZH1 or EZH2. The effect of miR-20a on HCC progression was studied in vivo and in vitro. The tumor inhibitory effect of UNC1999 was confirmed in vivo. CCK8 assay, wound healing assay, cell migration and invasion assay were used to evaluate the synergistic effect of UNC1999 with sorafenib. RNA sequencing (RNA-seq) was performed to screen the differentially expressed genes in the Huh7 and SMMC7721 cell lines after UNC1999, sorafenib, and combination treatments.ResultsIn this study, miR-20a showed a lower expression in both HCC tissues and cell lines. MiR-20a inhibited the proliferation and migration of SMMC7721 and Huh7 cells. The results of the luciferase assay and Western blot analysis revealed that miR-20a directly targeted EZH1, a histone methyltransferase. We demonstrated that miR-20a negatively regulated the expression of EZH1 and inhibited the proliferation and metastasis of HCC by reducing H3K27 methylation. We found UNC1999 inhibited tumor cells proliferation and enhanced the inhibitory effect of sorafenib.ConclusionWe demonstrated that miR-20a suppresses the tumor proliferation and metastasis in HCC by directly targeting EZH1. UNC1999 can inhibit tumor proliferation in vivo and increase the sensitivity of hepatoma cell lines to sorafenib.

Effect of Xiaoyutang Combined with Intraperitoneal Heat Perfusion Chemotherapy on Immune Function, Circulating mir, Prognosis, and Survival of Postoperative Patients with Colorectal Cancer.

Objective To explore the effects of Xiaoyutang combined with intraperitoneal heat perfusion chemotherapy on immune function, circulating Mir, and prognosis and survival of postoperative patients with colorectal cancer. Methods A total of 96 patients with colorectal cancer who were treated in our hospital from May 2018 to August 2019 and followed up to August 2021 were selected as the study subjects. The patients were randomly divided into a control group and study group by a 1 : 1 random number table method, 48 cases in each group. Patients in the control group were given intraperitoneal thermal perfusion chemotherapy after surgery, and patients in the research group were treated with Xiaoyutang on this basis. The treatment cycle was 21 days, and all patients were treated for 3 consecutive cycles. The therapeutic efficacy, immune function (CD3+, CD4+, and CD4+/CD8+), circulating mir (mir-29a, mir-145, and mir-92a), prognosis, and survival of the two groups were compared. Results After 3 cycles of treatment, ORR and DCR in the study group were higher than those in the control group (60.42% vs. 37.50%) and 85.42% vs. 66.67%, respectively, with statistical significance (P < 0.05). There were statistically significant differences in CD3+, CD4+, CD4+/CD8+, mir-29a, mir-145, and mir-92a time points and intergroup and intergroup interactions between the two groups (P < 0.05); the levels of CD3+, CD4+, and CD4+/CD8+ in the study group were higher than those in the control group after 1, 2, and 3 cycles of treatment (P < 0.05); the expressions of mir-29a, mir-145, and mir-92a were significantly lower than those in the control group (P < 0.05). By the end of follow-up, 3 cases were lost to follow-up in the study group and 5 cases in the control group. The recurrence rate and mortality of the study group were lower than those of the control group at 1- and 2-year follow-up (P > 0.05), and the mean survival time of patients in the study group was higher than that in the control group; the differences were statistically significant (χ2 = 5.151, P = 0.023). Conclusion Xiaoyutang combined with peritoneal heat perfusion chemotherapy has a good postoperative effect on patients with colorectal cancer, which can effectively improve the immune function and circulating Mir of patients with colorectal cancer, reduce tumor recurrence, and improve the prognosis of patients.

Effect of miRNA-200a on radiosensitivity of osteosarcoma cells by targeting Bone morphogenetic protein receptor 2

ABSTRACT To study the effect of miR-200a on radiosensitivity of osteosarcoma cells and its mechanism. NC (normal cell) group, mimic-NC group, mimic-miR-200a group, inhibitor-NC group, inhibitor-miR-200a group, si-NC group, si-BMPR2 (Bone morphogenetic protein receptor 2) group, mimic-miR-200a+vector-NC group, and mimic-miR-200a+vector-BMPR2 group were set; the cells of the above groups were irradiated with different radiation intensities (0, 1, 2, 3, and 4 Gy). The expression of miR-200a and BMPR2 mRNA was detected by qRT-PCR; the expression of BMPR2 protein was detected by Western blot; cell viability was detected by MMT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide); apoptosis rate was detected by flow cytometry. Cell clone formation experiment was used to detect cell radiosensitivity. Dual-luciferase reporter gene test was used to detect cell fluorescence activity. The expression of BMPR2 was high and the expression of miR-200a was low in osteosarcoma tissues after radiotherapy and in osteosarcoma cells after irradiation. Overexpression of miR-200a and interference with BMPR2 expression inhibits osteosarcoma cell proliferation, promotes apoptosis, and increases cellular radiosensitivity, miR-200a targets expression of BMPR2, and overexpression of BMPR2 reverses the radiosensitizing and apoptotic effects of miR-200a expression on osteosarcoma cells. Overexpression of miR-200a inhibits osteosarcoma cell proliferation, promotes apoptosis, and increases cellular radiosensitivity. The mechanism may be related to the regulation of BMPR2, which may provide new targets and new ideas for osteosarcoma treatment.

MicroRNA-141 and miR-200a induce the chondrogenic cell fate in human periodontal ligament cells by targeting TWIST2 and KLF12

Differentiation of mesenchymal stem cells (MSCs) in human periodontal ligaments (HPDLs) is induced by specific transcription factors that support periodontal tissue regeneration. The maintaining characteristics for HPDL cells could be regulated by two kinds of transcription factors, such as Twist family basic helix-loop-helix (bHLH) transcription factor 2 (TWIST2) and Krüppel-like factor 12 (KLF12). Because TWIST2 and KLF12 may be controlled by microRNAs (miRNAs), we investigated their 3′-untranslated regions (3′-UTRs) that contain seed sequences for miR-141 and miR-200a. HPDL cells were co-transfected with luciferase (LUC) reporter plasmids with or without 3′-UTRs for TWIST2 or KLF12, and miR-141 or miR-200a expression vectors. MiR-141 and miR-200a suppressed LUC activities via the TWIST2 or KLF12-3′-UTRs and reduced the mRNA and protein levels of TWIST2 and KLF12. These effects of miR-141 and miR-200a were related to reduced and increased expressions of the collagen Type I (COL1A1) and aggrecan (ACAN), while increasing the mRNAs for the chondrogenic factors SRY-Box transcription factor 5 (SOX5) SOX6, and Tricho-rhino-phalangeal syndrome type 1 (TRPS1) genes. We validated that these effects also reduce SOX5 and SOX6 proteins levels. Expression of miR-141 and miR-200a was significantly higher in HCS2/8 chondrogenic cells than human gingival fibroblasts (HGF), or HPDL and Saos2 cells. Furthermore, transfection of HPDL cells with miR-141 or miR-200a produced alcian blue staining. Thus, miR-141 and miR-200a directly target TWIST2 and KLF12 and induce chondrogenic differentiation of HPDL cells.

MicroRNA-106a regulates autophagy-related cell death and EMT by targeting TP53INP1 in lung cancer with bone metastasis

Bone metastasis is one of the most serious complications in lung cancer patients. MicroRNAs (miRNAs) play important roles in tumour development, progression and metastasis. A previous study showed that miR-106a is highly expressed in the tissues of lung adenocarcinoma with bone metastasis, but its mechanism remains unclear. In this study, we showed that miR-106a expression is dramatically increased in lung cancer patients with bone metastasis (BM) by immunohistochemical analysis. MiR-106a promoted A549 and SPC-A1 cell proliferation, migration and invasion in vitro. The results of bioluminescence imaging (BLI), micro-CT and X-ray demonstrated that miR-106a promoted bone metastasis of lung adenocarcinoma in vivo. Mechanistic investigations revealed that miR-106a upregulation promoted metastasis by targeting tumour protein 53-induced nuclear protein 1 (TP53INP1)-mediated metastatic progression, including cell migration, autophagy-dependent death and epithelial–mesenchymal transition (EMT). Notably, autophagy partially attenuated the effects of miR-106a on promoting bone metastasis in lung adenocarcinoma. These findings demonstrated that restoring the expression of TP53INP1 by silencing miR-106a may be a novel therapeutic strategy for bone metastatic in lung adenocarcinoma.

Differential Expression of micro RNAs and their Association with the Inflammatory Markers in Familial Mediterranean Fever Patients

Background: Familial Mediterranean fever (FMF) is an auto inflammatory genetic disease resulted from the mutation of pyrin, which contributes to the formation of inflamma some complex. Therefore, activation of cytokines is one of the hallmarks of FMF pathogenesis. This study aimed to investigate the role of miRNAs as regulatory biomarkers for inflammation in patients with FMF. Methods: 50 FMF patients and 25 healthy subjects were included in this study. Q RT-PCR was used to determine plasma expressions of miR-181a and miR-125a, while IFN-γ and IL-17 were estimated using ELISA technique. Results: Our results indicated that, the expression of miR-181a was significantly decreased (p = 0.006) while miR-125a expression was insignificantly reduced (p = 0.101) also IL-17 levels were significantly higher(p = 0.003) and plasma IFN-γ levels were insignificantly increased (p = 0.322) in FMF patients than control group. Correlation analysis revealed a positive correlation between miR-181a expression and lymphocyte percentages (p = 0.048),while a significant negative association was observed between miR-125a and C-reactive protein (CRP) (p = 0.005) in FMF patients. However, there were no associations between miR-125a and miR-181a with IFN-γ and IL-17 in FMF patients. Conclusion: miR-181a and miR-125a could be used as regulatory biomarkers for inflammation in FMF patients.

Long non-coding RNA KRT8P41/miR-193a-3p/FUBP1 axis modulates the proliferation and invasion of chordoma cells

Chordomas are low-grade malignancies accounting for 1–4% of primary bone malignancies. Moreover, local recurrences increase the rate of metastasis. Our previous study identified the far upstream element (FUSE)-binding protein 1 (FUBP1) as a biomarker and potential therapeutic target for chordoma. In this study, lncRNA KRT8P41 was identified as a lncRNA positively correlated with FUBP1. In chordoma patients, higher lncRNA KRT8P41 expression was correlated with a poorer prognosis. LncRNA KRT8P41 silencing significantly inhibited chordoma cell proliferation and invasion. miR-193a was negatively correlated with lncRNA KRT8P41 and FUBP1; lncRNA KRT8P41 inhibited miR-193a expression, and miR-193a inhibited FUBP1 expression. Furthermore, miR-193a directly bound to lncRNA KRT8P41 and FUBP1 and lncRNA KRT8P41 competed with FUBP1 for miR-193a binding and relieved miR-193a-mediated FUBP1 inhibition. LncRNA KRT8P41 silencing inhibited, whereas miR-193a inhibition promoted chordoma cell proliferation and invasion; the inhibition of miR-193a attenuated the roles of lncRNA KRT8P41. Within chordoma tissues, the expression of miR-193a was decreased, and the expression of FUBP1 increased compared to normal control tissues. LncRNA KRT8P41 exhibited a positive correlation with FUBP1 and a negative correlation with miR-193a in vivo. Therefore, it was concluded that lncRNA KRT8P41, miR-193a-3p, and FUBP1 form a lncRNA-miRNA-mRNA axis, modulating the proliferation and invasion of chordoma cells.