AIM:To identify molecular biologic differences between two gastric adenocarcinoma subgroups presenting different prognoses through the analysis of microRNA and protein expression.METHODS:Array technologies were used t...AIM:To identify molecular biologic differences between two gastric adenocarcinoma subgroups presenting different prognoses through the analysis of microRNA and protein expression.METHODS:Array technologies were used to generate1146 microRNAs and 124 proteins expression profiles of samples from 60 patients with gastric cancer.For the integrative analysis,we used established mRNA expression data published in our previous study.Whole mRNA expression levels were acquired from microarray data for 60 identical gastric cancer patients.Two gastric adenocarcinoma subgroups with distinct mRNA expression profiles presented distinctly different prognoses.MicroRNA and protein expression patterns were compared between gastric cancer tissue and normal gastric tissue and between two different prognostic groups.Aberrantly expressed microRNA,associated mRNA,and protein in patients with poor-prognosis gastric cancer were validated by quantitative reverse transcription polymerase chain reaction and immunochemistry in independent patients.RESULTS:We obtained the expression data of 1146microRNAs and 124 cancer-related proteins.Four microRNAs were aberrantly expressed in the two prognostic groups and in cancer vs non-cancer tissues(P<0.05).In the poor-prognosis group,miR-196b,miR-135b,and miR-93 were up-regulated and miR-29c*was down-regulated.miR-196b expression positively correlated with Homeobox A10(HOXA10)expression(r=0.726,P<0.001),which was significantly increased in poor-prognosis patients(P<0.001).Comparing gastric cancer with non-cancer tissues,46/124 proteins showed differential expression(P<0.05);COX2(P<0.001)and cyclin B1(P=0.017)were clearly overexpressed in the poor-prognosis group.CONCLUSION:Co-activation of miR-196b and HOXA10characterized a poor-prognosis subgroup of patients with gastric cancer.Elucidation of the biologic function of miR-196b and HOXA10 is warranted.展开更多
The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the r...The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the repair of damaged nerves.Previous studies showed that miR-221-3p promotes the proliferation and migration of SCs,and miR-338-3p promotes the myelination of SCs.In this study,we established rat models of sciatic nerve injury by bridging the transected sciatic nerve with a silicone tube.We injected a miR-221 lentiviral vector system together with a doxycycline-inducible Tet-On miR-338 lentiviral vector system into the cavity of nerve conduits of nerve stumps to sequentially regulate the biological function of endogenous SCs at different stages of nerve regeneration.We found that the biological function of SCs was sequentially regulated,the diameter and density of myelinated axons were increased,the expression levels of NF200 and myelin basic protein were increased,and the function of injured peripheral nerve was improved using this system.miRNA Target Prediction Database prediction,Nanopore whole transcriptome sequencing,quantitative PCR,and dual luciferase reporter gene assay results predicted and verified Cdkn1b and Nrp1 as target genes of miR-221-3p and miR-338-3p,respectively,and their regulatory effects on SCs were confirmed in vitro.In conclusion,here we established a new method to enhance nerve regeneration through sequential regulation of biological functions of endogenous SCs,which establishes a new concept and model for the treatment of peripheral nerve injury.The findings from this study will provide direct guiding significance for clinical treatment of sciatic nerve injury.展开更多
BACKGROUND Altered miR-188-3p expression has been observed in various human cancers.AIM To investigate the miR-188-3p expression,its roles,and underlying molecular events in gastric cancer.METHODS Fifty gastric cancer...BACKGROUND Altered miR-188-3p expression has been observed in various human cancers.AIM To investigate the miR-188-3p expression,its roles,and underlying molecular events in gastric cancer.METHODS Fifty gastric cancer and paired normal tissues were collected to analyze miR-188-3p and CBL expression.Normal and gastric cancer cells were used to manipulate miR-188-3p and CBL expression through different assays.The relationship between miR-188-3p and CBL was predicted bioinformatically and confirmed using a luciferase gene reporter assay.A Kaplan-Meier analysis was used to associate miR-188-3p or CBL expression with patient survival.A nude mouse tumor cell xenograft assay was used to confirm the in vitro data.RESULTS MiR-188-3p was found to be lower in the plasma of gastric cancer patients,tissues,and cell lines compared to their healthy counterparts.It was associated with overall survival of gastric cancer patients(P<0.001),tumor differentiation(P<0.001),lymph node metastasis(P=0.033),tumor node metastasis stage(I/II vs III/IV,P=0.024),and American Joint Committee on Cancer stage(I/II vs III/IV,P=0.03).Transfection with miR-188-3p mimics reduced tumor cell growth and invasion while inducing apoptosis and autophagy.CBL was identified as a direct target of miR-188-3p,with its expression antagonizing the effects of miR-188-3p on gastric cancer(GC)cell proliferation by inducing tumor cell apoptosis and autophagy through the inactivation of the Akt/mTOR signaling pathway.The in vivo data confirmed antitumor activity via CBL downregulation in gastric cancer.CONCLUSION The current data provides ex vivo,in vitro,and in vivo evidence that miR-188-3p acts as a tumor suppressor gene or possesses antitumor activity in GC.展开更多
Objective:To investigate the expression of microRNA-155(miR-155)in nasopharyngeal carcinoma and its effect on the proliferation of nasopharyngeal carcinoma cells.Methods:Patients with nasopharyngeal carcinoma who unde...Objective:To investigate the expression of microRNA-155(miR-155)in nasopharyngeal carcinoma and its effect on the proliferation of nasopharyngeal carcinoma cells.Methods:Patients with nasopharyngeal carcinoma who underwent surgical resection in our hospital from September 2016 to September 2018 were selected.miR-155 was detected by real-time fluorescence quantitative PCR in nasopharyngeal carcinoma tissues,normal tissues and nasopharyngeal carcinoma cell lines(CEN1),which were resected in our hospital from September 2016 to September 2018.The expression of NP69 in normal nasopharyngeal epithelial cells was detected by silencing miR-155,and its effect on the proliferation of human nasopharyngeal carcinoma cells was detected.Results:The expression of miR-155 in nasopharyngeal carcinoma tissue and cell line CEN1 was significantly higher than that in normal tissues.The expression level of NP69 in nasopharyngeal epithelial cells was significantly higher than that in nasopharyngeal epithelial cells(t=8.560,P=0.000;t=42.386,P=0.000).The expression of miR-155 was correlated with TNM stage,pathological grade,extent of invasion and lymph node metastasis in nasopharyngeal carcinoma(P<0.05),but not with age and gender(P>0.05).The expression of miR-155 after transfection of miR-155-inhibitor was significantly lower than that of the control group and the blank control group(F=35.63,P=0.003).The growth rate of nasopharyngeal carcinoma cells silenced by miR-155 was significantly slower than that of control group and blank control group(P<0.05).Conclusion:The high expression of miR-155 in human nasopharyngeal carcinoma tissues and cells can inhibit the proliferation of nasopharyngeal carcinoma cells,and it is of great significance for the treatment of nasopharyngeal carcinoma.展开更多
MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We in...MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We investigated miR-16 expression and their potential roles in a rat model of hypertrophy induced by abdominal artery constriction (AAC).miR-16 expression was significantly decreased, and CCND1 and CCND2 protein were markedly increased without obvious change of its mRNA level after hypertrophy induction.CCND1 and CCND2 levels were increased without changing their transcript levels in neonatal rat ventricular cardiomyocytes(NRVC) induced by PE,and miR-16 was down-regulated in this process with significantly up-regulatedβ-MHC,ANF and MLC-2 expression.Conversely,introduction of functional miR-16,CCND1 siRNA or CCND2 siRNA into NRVCs could repress cardiomyocyte hypertrophy.These results implicate that miR-16 is involved in contributing to cardiac hypertrophy,one of the mechanisms may be resulted from post-transcriptional regulation of CCND1 and CCND2.展开更多
Objective:To analyze the expression and clinical significance of miRNA-495 in the peripheral blood of patients with acute myeloid leukemia.Methods:Fifty-six patients with acute myeloid leukemia and 56 healthy controls...Objective:To analyze the expression and clinical significance of miRNA-495 in the peripheral blood of patients with acute myeloid leukemia.Methods:Fifty-six patients with acute myeloid leukemia and 56 healthy controls were selected.Fasting venous blood was drawn and centrifuged,and the plasma was collected.The target miRNA was directly amplified and reverse transcribed into cDNA.The expression of plasma miR-495 was detected by qRT-PCR.Results:The expression level of miRNA-495 in newly diagnosed acute myeloid leukemia(AML[ND])and relapsed/refractory acute myeloid leukemia(AML[RR])was significantly lower than that in complete remission(AML[CR])and normal control group(Control)(p<0.0001).There was no significant difference between AML(ND)group and AML(RR)group(p>0.05).The area under the ROC curve(AUC)of miRNA-495 was 0.9503,the 95%confidence interval was 0.9113–0.9892(p<0.0001),the standard error was 0.020,the sensitivity and specificity were 91.1%and 92.9%,respectively,and the Jordan index was 0.857.There was no significant difference between the expression level of miRNA-495 and gender,age,leukocyte count,hemoglobin,and platelet count(p>0.05).However,it was found related to the proportion of primitive bone marrow cells in patients(p=0.017).Conclusion:The decreased expression of plasma miR-495 in AML patients can be used as a new indicator for the diagnosis and prognosis of AML.展开更多
AIM: To determine if mir-30 d inhibits the autophagy response to Helicobacter pylori(H. pylori) invasion and increases H. pylori intracellular survival.METHODS: The expression of mir-30 d was detected by quantitative ...AIM: To determine if mir-30 d inhibits the autophagy response to Helicobacter pylori(H. pylori) invasion and increases H. pylori intracellular survival.METHODS: The expression of mir-30 d was detected by quantitative polymerase chain reaction(PCR), and autophagy level was examined by transmission electron microscopy, western blot, and GFP-LC3 puncta assay in human AGS cells and GES-1 cells. Luciferase reporter assay was applied to confirm the specificity of mir-30 d regulation on the expression of several core molecules involved in autophagy pathway. The expression of multiple core proteins were analyzed at both the m RNA and protein level, and the intracellular survival of H. pylori after different treatments was detected by gentamicin protection assay.RESULTS: Autophagy level was increased in AGS and GES-1 cells in response to H. pylori infection, which was accompanied by upregulation of mir-30 d expression(P < 0.05, vs no H. pylori infection). In the two gastric epithelial cell lines, mimic mir-30 d was found to repress the autophagy process, whereas mir-30 d inhibitor increased autophagy responseto H. pylori invasion. mir-30 d mimic decreased the luciferase activity of wild type reporter plasmids carrying the 3′ untranslated region(UTR) of all five tested genes(ATG2B, ATG5, ATG12, BECN1, and BNIP3L), whereas it had no effect on the mutant reporter plasmids. These five genes are core genes of autophagy pathway, and their expression was reduced significantly after mir-30 d mimic transfection(P < 0.05, vs control cells without mir-30 d mimic treatment). Mir-30 d mimic transfection and direct inhibition of autophagy increased the intracellular survival of H. pylori in AGS cells.CONCLUSION: Mir-30 d increases intracellular survival of H. pylori in gastric epithelial cells through inhibition of multiple core proteins in the autophagy pathway.展开更多
MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the ...MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically(P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased(P<0.001). In addition, the mRNA expression of miR-10b was negatively(P<0.01) correlated with DAZAP1 mRNA expression(r=–0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated(P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted(P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8(CCK-8) assay and the 5-Ethynyl-2′-deoxyuridine(EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect(P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted(P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.展开更多
MicroRNAs(miRNAs) can regulate the modulation of the phenotype of Schwann cells. Numerous novel miRNAs have been discovered and identified in rat sciatic nerve segments, including miR-3099. In the current study, miR-3...MicroRNAs(miRNAs) can regulate the modulation of the phenotype of Schwann cells. Numerous novel miRNAs have been discovered and identified in rat sciatic nerve segments, including miR-3099. In the current study, miR-3099 expression levels following peripheral nerve injury were measured in the proximal stumps of rat sciatic nerves after surgical crush. Real-time reverse transcription-polymerase chain reaction was used to determine miR-3099 expression in the crushed nerve segment at 0, 1, 4, 7, and 14 days post sciatic nerve injury, which was consistent with Solexa sequencing outcomes. Expression of miR-3099 was up-regulated following peripheral nerve injury. EdU and transwell chamber assays were used to observe the effect of miR-3099 on Schwann cell proliferation and migration. The results showed that increased miR-3099 expression promoted the proliferation and migration of Schwann cells. However, reduced miR-3099 expression suppressed the proliferation and migration of Schwann cells. The potential target genes of miR-3099 were also investigated by bioinformatic tools and high-throughput outcomes. miR-3099 targets genes Aqp4, St8 sia2, Tnfsf15, and Zbtb16 and affects the proliferation and migration of Schwann cells. This study examined the levels of miR-3099 at different time points following peripheral nerve injury. Our results confirmed that increased miR-3099 level induced by peripheral nerve injury can promote the proliferation and migration of Schwann cells.展开更多
BACKGROUND： The deficiency of liver regeneration needs to be addressed in the fields of liver surgery, split liver transplan- tation and living donor liver transplantation. Researches of microRNAs would broaden our u...BACKGROUND： The deficiency of liver regeneration needs to be addressed in the fields of liver surgery, split liver transplan- tation and living donor liver transplantation. Researches of microRNAs would broaden our understandings on the mecha- nisms of various diseases. Our previous research confirmed that miR-26a regulated liver regeneration in mice; however, the relationship between miR-26a and its target, directly or in- directly, remains unclear. Therefore, the present study further investigated the mechanism of miR-26a in regulating mouse hepatocyte proliferation. METHODS： An established mouse liver cell line, Nctc-1469, was transfected with Ad5-miR-26a-EGFP, Ad5-anti-miR-26a- EGFP or AdS-EGFP vector. Cell proliferation was assessed by MTS, cell apoptosis and cell cycle by flow cytometry, and gene expression by Western blotting and quantitative real-time PCR. Dual-luciferase reporter assays were used to test targets of miR-26a. RESULTS： Compared with the Ad5-EGFP group, Ad5-anti- miR-26a-EGFP down-regulated miR-26a and increased prolif- eration of hepatocytes, with more cells entering the G1 phase of cell cycle （82.70%＋1.45% vs 75.80%＋_3.92%）, and decreased apoptosis （5.50%＋0.35% vs 6.73%_＋0.42%）. CCND2 and CCNE2 were the direct targeted genes of miR-26a, miR-26a down- regulation up-regulated CCND2 and CCNE2 expressions and down-regulated p53 expression in Nctc-1469 cells. On the con- trary, miR-26a over-expression showed the opposite results. CONCLUSIONS： miR-26a regulated mouse hepatocyte pro- liferation by directly targeting the 3＇ untranslated regions of cyclin D2/cyclin E2; miR-26a also regulated p53-mediated apoptosis. Our data suggested that miR-26a may be a promis- ing regulator in liver regeneration.展开更多
MicroRNAs(miRNA)are recently discovered endogenous,small noncoding RNAs(of 22 nucleotides)that play pivotal roles in gene regulation.They are involved in post-transcriptional control of gene expression.miRNAs are emer...MicroRNAs(miRNA)are recently discovered endogenous,small noncoding RNAs(of 22 nucleotides)that play pivotal roles in gene regulation.They are involved in post-transcriptional control of gene expression.miRNAs are emerging as important regulators of cell proliferation,development,cancer formation,stress responses,cell death and physiological conditions.Increasing evidence has demonstrated the human miRNAs bind to their target mRNA sequences with perfect or near-perfect sequence complementarily.This provides a powerful strategy for discovering potential type 2 diabetes mellitus(T2DM)targets and gives the probability to exploit them for diagnostic and therapeutic causes.About 6%of the world population is affected by T2DM,and it is recognized as a global epidemic by the World Health Organization.At present there is no valid biomarker to control or manage T2DM.Therefore,the present study applied a mature sequence of miRNAs from publicly accessible databases to identify the miRNA from T2DM expressed sequence tags,and the results are detailed and discussed below.展开更多
基金Supported by The Faculty Research Grant of Yonsei University College of Medicine(6-2011-0113)the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education,Science and Technology,No.2010-0024248
文摘AIM:To identify molecular biologic differences between two gastric adenocarcinoma subgroups presenting different prognoses through the analysis of microRNA and protein expression.METHODS:Array technologies were used to generate1146 microRNAs and 124 proteins expression profiles of samples from 60 patients with gastric cancer.For the integrative analysis,we used established mRNA expression data published in our previous study.Whole mRNA expression levels were acquired from microarray data for 60 identical gastric cancer patients.Two gastric adenocarcinoma subgroups with distinct mRNA expression profiles presented distinctly different prognoses.MicroRNA and protein expression patterns were compared between gastric cancer tissue and normal gastric tissue and between two different prognostic groups.Aberrantly expressed microRNA,associated mRNA,and protein in patients with poor-prognosis gastric cancer were validated by quantitative reverse transcription polymerase chain reaction and immunochemistry in independent patients.RESULTS:We obtained the expression data of 1146microRNAs and 124 cancer-related proteins.Four microRNAs were aberrantly expressed in the two prognostic groups and in cancer vs non-cancer tissues(P<0.05).In the poor-prognosis group,miR-196b,miR-135b,and miR-93 were up-regulated and miR-29c*was down-regulated.miR-196b expression positively correlated with Homeobox A10(HOXA10)expression(r=0.726,P<0.001),which was significantly increased in poor-prognosis patients(P<0.001).Comparing gastric cancer with non-cancer tissues,46/124 proteins showed differential expression(P<0.05);COX2(P<0.001)and cyclin B1(P=0.017)were clearly overexpressed in the poor-prognosis group.CONCLUSION:Co-activation of miR-196b and HOXA10characterized a poor-prognosis subgroup of patients with gastric cancer.Elucidation of the biologic function of miR-196b and HOXA10 is warranted.
基金supported by the National Natural Science Foundation of China,No.81771351the National Key R&D Program of China,No.2017YFA0105802+1 种基金the Joint Research Fund Liaoning-Shenyang National Laboratory for Materials Science,No.2019JH3/30100022the National Science Foundation for Post-doctoral Scientists of China,No.2018M641732(all to QA and LLW)。
文摘The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the repair of damaged nerves.Previous studies showed that miR-221-3p promotes the proliferation and migration of SCs,and miR-338-3p promotes the myelination of SCs.In this study,we established rat models of sciatic nerve injury by bridging the transected sciatic nerve with a silicone tube.We injected a miR-221 lentiviral vector system together with a doxycycline-inducible Tet-On miR-338 lentiviral vector system into the cavity of nerve conduits of nerve stumps to sequentially regulate the biological function of endogenous SCs at different stages of nerve regeneration.We found that the biological function of SCs was sequentially regulated,the diameter and density of myelinated axons were increased,the expression levels of NF200 and myelin basic protein were increased,and the function of injured peripheral nerve was improved using this system.miRNA Target Prediction Database prediction,Nanopore whole transcriptome sequencing,quantitative PCR,and dual luciferase reporter gene assay results predicted and verified Cdkn1b and Nrp1 as target genes of miR-221-3p and miR-338-3p,respectively,and their regulatory effects on SCs were confirmed in vitro.In conclusion,here we established a new method to enhance nerve regeneration through sequential regulation of biological functions of endogenous SCs,which establishes a new concept and model for the treatment of peripheral nerve injury.The findings from this study will provide direct guiding significance for clinical treatment of sciatic nerve injury.
基金Supported by the National Natural Science Funds of China,No.81974448Guangdong Medical Research Foundation,No.B2019126Shenzhen Science and Technology Innovation Commission,No.JCYJ20210324135005013.
文摘BACKGROUND Altered miR-188-3p expression has been observed in various human cancers.AIM To investigate the miR-188-3p expression,its roles,and underlying molecular events in gastric cancer.METHODS Fifty gastric cancer and paired normal tissues were collected to analyze miR-188-3p and CBL expression.Normal and gastric cancer cells were used to manipulate miR-188-3p and CBL expression through different assays.The relationship between miR-188-3p and CBL was predicted bioinformatically and confirmed using a luciferase gene reporter assay.A Kaplan-Meier analysis was used to associate miR-188-3p or CBL expression with patient survival.A nude mouse tumor cell xenograft assay was used to confirm the in vitro data.RESULTS MiR-188-3p was found to be lower in the plasma of gastric cancer patients,tissues,and cell lines compared to their healthy counterparts.It was associated with overall survival of gastric cancer patients(P<0.001),tumor differentiation(P<0.001),lymph node metastasis(P=0.033),tumor node metastasis stage(I/II vs III/IV,P=0.024),and American Joint Committee on Cancer stage(I/II vs III/IV,P=0.03).Transfection with miR-188-3p mimics reduced tumor cell growth and invasion while inducing apoptosis and autophagy.CBL was identified as a direct target of miR-188-3p,with its expression antagonizing the effects of miR-188-3p on gastric cancer(GC)cell proliferation by inducing tumor cell apoptosis and autophagy through the inactivation of the Akt/mTOR signaling pathway.The in vivo data confirmed antitumor activity via CBL downregulation in gastric cancer.CONCLUSION The current data provides ex vivo,in vitro,and in vivo evidence that miR-188-3p acts as a tumor suppressor gene or possesses antitumor activity in GC.
基金This study was supported by Guangdong Medical Research Fund Project(Grant No.A2014870).
文摘Objective:To investigate the expression of microRNA-155(miR-155)in nasopharyngeal carcinoma and its effect on the proliferation of nasopharyngeal carcinoma cells.Methods:Patients with nasopharyngeal carcinoma who underwent surgical resection in our hospital from September 2016 to September 2018 were selected.miR-155 was detected by real-time fluorescence quantitative PCR in nasopharyngeal carcinoma tissues,normal tissues and nasopharyngeal carcinoma cell lines(CEN1),which were resected in our hospital from September 2016 to September 2018.The expression of NP69 in normal nasopharyngeal epithelial cells was detected by silencing miR-155,and its effect on the proliferation of human nasopharyngeal carcinoma cells was detected.Results:The expression of miR-155 in nasopharyngeal carcinoma tissue and cell line CEN1 was significantly higher than that in normal tissues.The expression level of NP69 in nasopharyngeal epithelial cells was significantly higher than that in nasopharyngeal epithelial cells(t=8.560,P=0.000;t=42.386,P=0.000).The expression of miR-155 was correlated with TNM stage,pathological grade,extent of invasion and lymph node metastasis in nasopharyngeal carcinoma(P<0.05),but not with age and gender(P>0.05).The expression of miR-155 after transfection of miR-155-inhibitor was significantly lower than that of the control group and the blank control group(F=35.63,P=0.003).The growth rate of nasopharyngeal carcinoma cells silenced by miR-155 was significantly slower than that of control group and blank control group(P<0.05).Conclusion:The high expression of miR-155 in human nasopharyngeal carcinoma tissues and cells can inhibit the proliferation of nasopharyngeal carcinoma cells,and it is of great significance for the treatment of nasopharyngeal carcinoma.
文摘MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We investigated miR-16 expression and their potential roles in a rat model of hypertrophy induced by abdominal artery constriction (AAC).miR-16 expression was significantly decreased, and CCND1 and CCND2 protein were markedly increased without obvious change of its mRNA level after hypertrophy induction.CCND1 and CCND2 levels were increased without changing their transcript levels in neonatal rat ventricular cardiomyocytes(NRVC) induced by PE,and miR-16 was down-regulated in this process with significantly up-regulatedβ-MHC,ANF and MLC-2 expression.Conversely,introduction of functional miR-16,CCND1 siRNA or CCND2 siRNA into NRVCs could repress cardiomyocyte hypertrophy.These results implicate that miR-16 is involved in contributing to cardiac hypertrophy,one of the mechanisms may be resulted from post-transcriptional regulation of CCND1 and CCND2.
基金supported by the Department of Science and Technology of Shaanxi Province(Grant Number:2017JM8160).
文摘Objective:To analyze the expression and clinical significance of miRNA-495 in the peripheral blood of patients with acute myeloid leukemia.Methods:Fifty-six patients with acute myeloid leukemia and 56 healthy controls were selected.Fasting venous blood was drawn and centrifuged,and the plasma was collected.The target miRNA was directly amplified and reverse transcribed into cDNA.The expression of plasma miR-495 was detected by qRT-PCR.Results:The expression level of miRNA-495 in newly diagnosed acute myeloid leukemia(AML[ND])and relapsed/refractory acute myeloid leukemia(AML[RR])was significantly lower than that in complete remission(AML[CR])and normal control group(Control)(p<0.0001).There was no significant difference between AML(ND)group and AML(RR)group(p>0.05).The area under the ROC curve(AUC)of miRNA-495 was 0.9503,the 95%confidence interval was 0.9113–0.9892(p<0.0001),the standard error was 0.020,the sensitivity and specificity were 91.1%and 92.9%,respectively,and the Jordan index was 0.857.There was no significant difference between the expression level of miRNA-495 and gender,age,leukocyte count,hemoglobin,and platelet count(p>0.05).However,it was found related to the proportion of primitive bone marrow cells in patients(p=0.017).Conclusion:The decreased expression of plasma miR-495 in AML patients can be used as a new indicator for the diagnosis and prognosis of AML.
基金Supported by the National Natural Science Fund from China,No.81260326
文摘AIM: To determine if mir-30 d inhibits the autophagy response to Helicobacter pylori(H. pylori) invasion and increases H. pylori intracellular survival.METHODS: The expression of mir-30 d was detected by quantitative polymerase chain reaction(PCR), and autophagy level was examined by transmission electron microscopy, western blot, and GFP-LC3 puncta assay in human AGS cells and GES-1 cells. Luciferase reporter assay was applied to confirm the specificity of mir-30 d regulation on the expression of several core molecules involved in autophagy pathway. The expression of multiple core proteins were analyzed at both the m RNA and protein level, and the intracellular survival of H. pylori after different treatments was detected by gentamicin protection assay.RESULTS: Autophagy level was increased in AGS and GES-1 cells in response to H. pylori infection, which was accompanied by upregulation of mir-30 d expression(P < 0.05, vs no H. pylori infection). In the two gastric epithelial cell lines, mimic mir-30 d was found to repress the autophagy process, whereas mir-30 d inhibitor increased autophagy responseto H. pylori invasion. mir-30 d mimic decreased the luciferase activity of wild type reporter plasmids carrying the 3′ untranslated region(UTR) of all five tested genes(ATG2B, ATG5, ATG12, BECN1, and BNIP3L), whereas it had no effect on the mutant reporter plasmids. These five genes are core genes of autophagy pathway, and their expression was reduced significantly after mir-30 d mimic transfection(P < 0.05, vs control cells without mir-30 d mimic treatment). Mir-30 d mimic transfection and direct inhibition of autophagy increased the intracellular survival of H. pylori in AGS cells.CONCLUSION: Mir-30 d increases intracellular survival of H. pylori in gastric epithelial cells through inhibition of multiple core proteins in the autophagy pathway.
基金financially supported by the earmarked fund for China Agriculture Research System (CARS-36)the Hunan Provincial Natural Science Foundation of China (2018JJ2176 and 2018JJ3219)
文摘MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically(P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased(P<0.001). In addition, the mRNA expression of miR-10b was negatively(P<0.01) correlated with DAZAP1 mRNA expression(r=–0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated(P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted(P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8(CCK-8) assay and the 5-Ethynyl-2′-deoxyuridine(EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect(P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted(P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.
基金supported by the Postgraduate Research&Practice Innovation Program of Jiangsu Province of China,No.KYCX17-1910(to QYL)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions of China(PAPD)
文摘MicroRNAs(miRNAs) can regulate the modulation of the phenotype of Schwann cells. Numerous novel miRNAs have been discovered and identified in rat sciatic nerve segments, including miR-3099. In the current study, miR-3099 expression levels following peripheral nerve injury were measured in the proximal stumps of rat sciatic nerves after surgical crush. Real-time reverse transcription-polymerase chain reaction was used to determine miR-3099 expression in the crushed nerve segment at 0, 1, 4, 7, and 14 days post sciatic nerve injury, which was consistent with Solexa sequencing outcomes. Expression of miR-3099 was up-regulated following peripheral nerve injury. EdU and transwell chamber assays were used to observe the effect of miR-3099 on Schwann cell proliferation and migration. The results showed that increased miR-3099 expression promoted the proliferation and migration of Schwann cells. However, reduced miR-3099 expression suppressed the proliferation and migration of Schwann cells. The potential target genes of miR-3099 were also investigated by bioinformatic tools and high-throughput outcomes. miR-3099 targets genes Aqp4, St8 sia2, Tnfsf15, and Zbtb16 and affects the proliferation and migration of Schwann cells. This study examined the levels of miR-3099 at different time points following peripheral nerve injury. Our results confirmed that increased miR-3099 level induced by peripheral nerve injury can promote the proliferation and migration of Schwann cells.
基金supported by grants from the Key Clinical Project from the Ministry of Health(159)the National Natural Science Foundation of China(30972951 and 81170448)+1 种基金Special Fund for Science Research by Ministry of Health(201002004)the PhD Programs Foundation of Ministry of Education of China(20130171120076)
文摘BACKGROUND： The deficiency of liver regeneration needs to be addressed in the fields of liver surgery, split liver transplan- tation and living donor liver transplantation. Researches of microRNAs would broaden our understandings on the mecha- nisms of various diseases. Our previous research confirmed that miR-26a regulated liver regeneration in mice; however, the relationship between miR-26a and its target, directly or in- directly, remains unclear. Therefore, the present study further investigated the mechanism of miR-26a in regulating mouse hepatocyte proliferation. METHODS： An established mouse liver cell line, Nctc-1469, was transfected with Ad5-miR-26a-EGFP, Ad5-anti-miR-26a- EGFP or AdS-EGFP vector. Cell proliferation was assessed by MTS, cell apoptosis and cell cycle by flow cytometry, and gene expression by Western blotting and quantitative real-time PCR. Dual-luciferase reporter assays were used to test targets of miR-26a. RESULTS： Compared with the Ad5-EGFP group, Ad5-anti- miR-26a-EGFP down-regulated miR-26a and increased prolif- eration of hepatocytes, with more cells entering the G1 phase of cell cycle （82.70%＋1.45% vs 75.80%＋_3.92%）, and decreased apoptosis （5.50%＋0.35% vs 6.73%_＋0.42%）. CCND2 and CCNE2 were the direct targeted genes of miR-26a, miR-26a down- regulation up-regulated CCND2 and CCNE2 expressions and down-regulated p53 expression in Nctc-1469 cells. On the con- trary, miR-26a over-expression showed the opposite results. CONCLUSIONS： miR-26a regulated mouse hepatocyte pro- liferation by directly targeting the 3＇ untranslated regions of cyclin D2/cyclin E2; miR-26a also regulated p53-mediated apoptosis. Our data suggested that miR-26a may be a promis- ing regulator in liver regeneration.
文摘MicroRNAs(miRNA)are recently discovered endogenous,small noncoding RNAs(of 22 nucleotides)that play pivotal roles in gene regulation.They are involved in post-transcriptional control of gene expression.miRNAs are emerging as important regulators of cell proliferation,development,cancer formation,stress responses,cell death and physiological conditions.Increasing evidence has demonstrated the human miRNAs bind to their target mRNA sequences with perfect or near-perfect sequence complementarily.This provides a powerful strategy for discovering potential type 2 diabetes mellitus(T2DM)targets and gives the probability to exploit them for diagnostic and therapeutic causes.About 6%of the world population is affected by T2DM,and it is recognized as a global epidemic by the World Health Organization.At present there is no valid biomarker to control or manage T2DM.Therefore,the present study applied a mature sequence of miRNAs from publicly accessible databases to identify the miRNA from T2DM expressed sequence tags,and the results are detailed and discussed below.
