5,723 research outputs found
miR-127 protects proximal tubule cells against ischemia/reperfusion : identification of Kinesin family member 3B as miR-127 target
No full textIschemia/reperfusion (I/R) is at the basis of renal transplantation and acute kidney injury. Molecular mechanisms underlying proximal tubule response to I/R will allow the identification of new therapeutic targets for both clinical settings. microRNAs have emerged as crucial and tight regulators of the cellular response to insults including hypoxia. Here, we have identified several miRNAs involved in the response of the proximal tubule cell to I/R. Microarrays and RT-PCR analysis of proximal tubule cells submitted to I/R mimicking conditions in vitro demonstrated that miR-127 is induced during ischemia and also during reperfusion. miR-127 is also modulated in a rat model of renal I/R. Interference approaches demonstrated that ischemic induction of miR-127 is mediated by Hypoxia Inducible Factor-1alpha (HIF-1α) stabilization. Moreover, miR-127 is involved in cell-matrix and cell-cell adhesion maintenance, since overexpression of miR-127 maintains focal adhesion complex assembly and the integrity of tight junctions. miR-127 also regulates intracellular trafficking since miR-127 interference promotes dextran-FITC uptake. In fact, we have identified the Kinesin Family Member 3B (KIF3B), involved in cell trafficking, as a target of miR-127 in rat proximal tubule cells. In summary, we have described a novel role of miR-127 in cell adhesion and its regulation by HIF-1α. We also identified for the first time KIF3B as a miR-127 target. Both, miR-127 and KIF3B appear as key mediators of proximal epithelial tubule cell response to I/R with potential al application in renal ischemic damage management
miR-132/212 knockout mice reveal roles for these miRNAs in regulating cortical synaptic transmission and plasticity
No full textmiR-132 and miR-212 are two closely related miRNAs encoded in the same intron of a small non-coding gene, which have been suggested to play roles in both immune and neuronal function. We describe here the generation and initial characterisation of a miR-132/212 double knockout mouse. These mice were viable and fertile with no overt adverse phenotype. Analysis of innate immune responses, including TLR-induced cytokine production and IFNβ induction in response to viral infection of primary fibroblasts did not reveal any phenotype in the knockouts. In contrast, the loss of miR-132 and miR-212, while not overtly affecting neuronal morphology, did affect synaptic function. In both hippocampal and neocortical slices miR-132/212 knockout reduced basal synaptic transmission, without affecting paired-pulse facilitation. Hippocampal long-term potentiation (LTP) induced by tetanic stimulation was not affected by miR-132/212 deletion, whilst theta burst LTP was enhanced. In contrast, neocortical theta burst-induced LTP was inhibited by loss of miR-132/212. Together these results indicate that miR-132 and/or miR-212 play a significant role in synaptic function, possibly by regulating the number of postsynaptic AMPA receptors under basal conditions and during activity-dependent synaptic plasticity
Bta-miR-615 analysis in blastocysts.
No full textBta-miR-615 relative expression miRNA in blastocysts cultured conventionally (control) or in LM system. Statistics was performed using Student’s t test. Asterisk means statistical difference (P<0.05). Standard bars represents the SEM.</p
Colección: Perfil #3
No full textThis board-book version of LM turns out to be quite creative. Ratoncete comes from school every afternoon and goes through the forest looking for adventures. He apparently blasts a horn into the ear of the sleeping lion. Don Leon wants to spank him as a result, but Ratoncete offers an apology, not an offer of help. Later, he happens upon the lion in his trap of ropes. 8 pages, counting both covers. 6½" x 9".Language note: SpanishNo Autho
Identification and validation of oncologic miRNA biomarkers for Luminal A-like breast cancer
No full textIntroduction: Breast cancer is a common disease with distinct tumor subtypes phenotypically characterized by ER and HER2/neu receptor status. MiRNAs play regulatory roles in tumor initiation and progression, and altered miRNA expression has been demonstrated in a variety of cancer states presenting the potential for exploitation as cancer biomarkers. Blood provides an excellent medium for biomarker discovery. This study investigated systemic miRNAs differentially expressed in Luminal A-like (ER+PR+HER2/neu-) breast cancer and their effectiveness as oncologic biomarkers in the clinical setting. Methods: Blood samples were prospectively collected from patients with Luminal A-like breast cancer (n=54) and controls (n=56). RNA was extracted, reverse transcribed and subjected to microarray analysis (n=10 Luminal A-like; n=10 Control). Differentially expressed miRNAs were identified by artificial neural network (ANN) data-mining algorithms. Expression of specific miRNAs was validated by RQ-PCR (n=44 Luminal A; n=46 Control) and potential relationships between circulating miRNA levels and clinicopathological features of breast cancer were investigated. Results: Microarray analysis identified 76 differentially expressed miRNAs. ANN revealed 10 miRNAs for further analysis ( miR-19b, miR-29a, miR-93, miR-181a, miR-182, miR-223, miR-301a, miR-423-5p, miR-486-5 and miR-652 ). The biomarker potential of 4 miRNAs ( miR-29a, miR-181a , miR-223 and miR-652 ) was confirmed by RQ-PCR, with significantly reduced expression in blood of women with Luminal A-like breast tumors compared to healthy controls (p=0.001, 0.004, 0.009 and 0.004 respectively). Binary logistic regression confirmed that combination of 3 of these miRNAs ( miR-29a, miR-181a and miR-652 ) could reliably differentiate between cancers and controls with an AUC of 0.80. Conclusion: This study provides insight into the underlying molecular portrait of Luminal A-like breast cancer subtype. From an initial 76 miRNAs, 4 were validated with altered expression in the blood of women with Luminal A-like breast cancer. The expression profiles of these 3 miRNAs, in combination with mammography, has potential to facilitate accurate subtype- specific breast tumor detection
Could let-7f, miR-10b, miR-34a, miR-181b, and miR-181d Be Useful Tools as a Target Therapy for Uterine Leiomyosarcoma?
No full textBackground/Objectives: We have previously identified let-7f-5p, miR-10b-5p, miR-34a-5p, miR-181b-5p, and miR-181d-5p as differentially expressed between uterine leiomyoma (LM) and leiomyosarcoma (LMS) tissue samples. The present study aimed to characterize these miRNA expression profiles and to assess the functional role of miR-34a and miR-181b in uterine LM and LMS cells. Methods: All the selected miRNAs showed downregulation in LMS cells compared to LM cells, but only miR-34a and miR-181b expression patterns matched those of patient samples. Therefore, these two miRs were selected for further analyses. Results: Loss of function analysis demonstrated that miR-34a and miR-181b silencing inhibited LM cell proliferation and migration. MiR-34a silencing induced CCND1 and MDM4 expression and inhibited KMT2D, BCL2, and NOTCH2 in LM. Silencing of miR-181b promotes TIMP3 and FGFR1 expression in LM and diminishes BCL2, NOTCH2, ATM, IRS1, and PRLR. Gain of function analysis revealed that the introduction of miR-34a and miR-181b mimics suppressed proliferation and migration in malignant LMS cells. Additionally, transfection with a miR-34a mimic downregulated NOTCH2 and BCL2 expression and enhanced the expression of CCND1, KMT2D, and TP53 in LMS cells. Moreover, miR-181b overexpression decreased TIMP3, NOTCH2, ATM, and IRS1 expression and increased the expression of FGFR1 in this cell. Importantly, the single introduction of either a miR-34a or miR-181b mimic was able to decrease the invasion capacity of LMS cells. Conclusions: Our studies demonstrated that miR-34a or miR-181b may play an anti-oncogenic role in uterine tumors; further studies are needed to better understand the role and regulatory mechanism of these miRNAs in LMS cancer development, which will help provide prognostic and therapeutic options for patients with LMS
miR-21, miR-155, miR-192, and miR-375 Deregulations Related to NF-kappaB Activation in Gastroduodenal Fluid–Induced Early Preneoplastic Lesions of Laryngeal Mucosa In Vivo
No full textGastroduodenal refluxate found in the upper aerodigestive tract is not clinically uncommon. We recently demonstrated the neoplastic potential of gastroduodenal fluids (GDF) on hypopharyngeal mucosa, via NF-κB, using in vitro and in vivo models. Here we will explore the in vivo effect of GDF on laryngeal mucosa (LM) to induce early preneoplastic lesions related to NF-κB activation, along with deregulation of specific microRNA (miRNA) markers previously linked to laryngeal cancer. We used histological, immunohistochemical, automated quantitative analysis and quantitative polymerase chain reaction to examine LM from 35 C57Bl/6J mice previously treated with topical GDF against corresponding controls (4 experimental and 3 control groups; 5 mice/group). Our analysis showed that GDF produced early preneoplastic lesions in treated LM related to NF-κB activation. LM treated by acid and bile combination demonstrated significantly higher expression of the analyzed cell proliferation markers (Ki67, CK14, ∆Np63), oncogenic p-STAT3, and changes of cell adhesion molecules (E-cadherin, ϐ-catenin) versus untreated LM or LM exposed to acid alone (P < .0005). Furthermore, acidic bile but not neutral bile appeared to accelerate the expression of “oncomirs” miR-21, miR-155, and miR-192 (acidic bile versus neutral bile, P < .0001), while reducing tumor suppressor miR-375 (acidic bile versus neutral bile, P = .0137), previously linked to NF-κB and laryngeal cancer. Finally, acidic bile induced reduction of miR-34a, miR-375, and miR-451a, exhibiting an inverse correlation with NF-κB activation. SIGNIFICANCE: Bile in combination with acid has a selective tumorigenic effect on LM, inducing deregulation of “oncomirs” and tumor suppressor miRNAs, produced by NF-κB activation with molecular and early histopathological alterations linked to neoplastic transformation. Systematic acid suppression may in part convey a protective role
Functional analysis of let-7f-5p, miR-10b-5p, miR-34a-5p, miR-181b-5p and miR-181d-5p miRNAs in uterine leiomyoma and leiomyosarcoma cells
No full textA expressão anormal dos miRNAs está diretamente associada ao desenvolvimento de neoplasias por regularem a expressão de genes supressores tumorais e oncogenes. Assim, sua desregulação está intimamente relacionada a um pior prognóstico e comprometimento da resposta terapêutica. Embora esteja bem estabelecido o papel dos miRNAs em diversos tipos de cânceres, pouco se sabe sobre sua função nos tumores de músculo liso uterinos. Os leiomiossarcomas (LMS) são tumores raros com altas taxas de mortalidade e recorrência, enquanto os leiomiomas (LM) são tumores frequentes com ampla variabilidade histológica. A origem desses tumores é incerta, sendo controversa a possível malignização de um LM, e seu diagnóstico diferencial ainda representa grande desafio na pratica médica. Nosso grupo, em trabalho anterior, identificou uma série de miRNAs associados ao desenvolvimento de tumores com expressão diferencial entre LM e LMS uterinos. O principal objetivo desta tese foi selecionar e caracterizar miRNAs que pudessem ser aplicáveis para fins de diagnóstico diferencial, predição de prognóstico e resposta à terapêutica, ou ainda potenciais alvos para terapia específica nesses tumores. Cinco miRNAs (let-7f-5p, miR-10b-5p, miR-34a-5p, miR-181b-5p e miR181d-5p) foram selecionados para análise e caraterização em amostras de pacientes e linhagens celulares. A partir desses resultados, miR-34a e miR-181b foram selecionados para análise dos efeitos de sua indução (Mimic) e inibição (siRNA) in vitro. Após a manipulação molecular desses miRNAs, foram avaliadas a proliferação e migração celular, bem como a expressão de alguns de seus genes alvo (CCND1, MDM4, TP53 e BCL2 para o miR-34a; e TIMP3, FGFR1, ESR1 e BCL2 para o miR-181b). Nossos resultados mostraram associação entre perda de expressão de let-7f, miR-10b, miR-34a e miR-181d e uma pior sobrevida global das pacientes. A expressão de let-7f e miR-181d foi associada a menor sobrevida livre de doença. A transfecção das células com miR-34a-Mimic e -siRNA resultou em menor proliferação e migração das duas linhagens. miR-181b-Mimic induziu menor proliferação e migração no LM, enquanto o siRNA afetou somente a proliferação. No LMS, a proliferação foi inibida somente com o siRNA, enquanto a migração foi menor após transfecção com o miR-181b-Mimic e siRNA. Adicionalmente, miR-34a parece regular a expressão de CCND1, MDM4, TP53 e BCL2, nos LMS, uma vez que sua maior expressão coincidiu com menores níveis de CCND1 e sua inibição resultou no aumento de MDM4, TP53 e BCL2. A inibição do miR-181b levou a maior expressão de ESR1, no LM, e de BCL2, no LMS. Embora estudos mais detalhados sejam necessários para definir a utilização de miR-34a e miR-181b para fins de diagnóstico, prognostico e alvo terapêutico nesses tumores, nosso estudo demonstrou um potencial promissor dessas moléculas como biomarcadores moleculares no LMS uterino.Abnormal expression of miRNAs is directly associated with neoplasias development because they regulate the expression of tumor suppressor and oncogenes. Thereby, their deregulation are closely related to a worse prognosis and impairment of treatment response. Although the role of the miRNAs is well established in several cancer types, little is known about their function in uterine smooth muscle tumors. Leiomyosarcomas (LMS) are rare tumors with high rates of mortality and recurrence, while the leiomyoma (LM) are frequent tumors with wide histological variability. The origin of these tumors is uncertain, being controversial the possibility of malignancy of a LM, and its differential diagnosis represents a great challenge in medical practice yet. Our group, in a previous study identified several miRNAs associated with tumors development with differential expression between the LM and LMS. The main aim of this thesis was to select and characterize miRNAs that might be useful for differential diagnosis, prognostic prediction, and therapeutic response, or potential targets for specific therapy in these tumors. Five miRNAs (let-7f-5p, miR-10b-5p, miR-34a-5p, miR-181b-5p and miR-181d-5p) were selected for characterization and assessment in patients samples and cell lines. Based on these results, the miR-34a and miR-181b were selected for in vitro analysis of induction (Mimic) and inhibition (siRNA) of their expressions. After the molecular manipulation of these miRNAs, the cell proliferation and migration were evaluated, as well as the expression of some target-genes (CCND1, MDM4, TP53 and BCL2 for miR-34a; and TIMP3, FGFR1, ESR1 and BCL2 for miR-181b). Our results showed an association between loss expression of let-7f, miR-10b, miR-34a, and miR-181d and a worse overall survival of patients. Expression of let-7f and miR-181d were associated with the lowest disease-free survival. Cell tranfection with miR-34a Mimic and siRNA led to a decrease of proliferation and migration in both cell lines. miR-181b Mimic induced a decrease of proliferation and migration in LM, while the siRNA affected only the cell proliferation. In the LMS, the proliferation was inhibited only with the siRNA, while the cell migration was lower after miR-181b Mimic and siRNA. Additionally, miR-34a seems to regulate the CCND1, MDM4, TP53 and BCL2 expressions in LMS, since its greater expression corresponded with the lowest levels of CCND1, and its inhibition led to the increased of MDM4, TP53, and BCL2. Inhibition of miR-181b led to the greater expression of ESR1 in LM, and BCL2 in LMS. Although more detailed studies are necessary to define the use of miR-34a and miR-181b for diagnostic purposes, prognostic and therapeutic target in these tumors, our study demonstrated the promising potential of these molecules as molecular biomarkers in uterine LMS
<i>In vitro</i> and <i>in vivo</i> studies for miR-106b.
No full text<p>miR-106b was over-expressed in PLC-PT and knock-down in PLC-LM cells. (A) Wound-healing assay for miR-106b knock-down using PLC-LM cells and (B) miR-106b over-expression using PLC-PT cells. Higher miR-106b expressing cells, PLC-LM/Scr and PLC-PT/106b+, can promote cell migration <i>in vitro</i>. (C) Stress fiber (white arrows) protruding out from cell surface in high miR-106b expressing cells, PLC-LM/LNA-Scr, but not in the miR-106b knock-down cells, PLC-LM/LNA-106b. (D) Similar founding was observed in miR-106b over-expressing experiment. (E) H&E staining for <i>in vivo</i> studies showing lung metastatic nodules were present in miR-106b over-expressing cell, PLC-PT/106b+.</p
Int J Cancer
No full textBladder cancer is the fourth most common cancer among men in the United States and more than half of patients experience recurrences within 5 years after initial diagnosis. Additional clinically informative and actionable biomarkers of the recurrent bladder cancer phenotypes are needed to improve screening and molecular therapeutic approaches for recurrence prevention. MicroRNA-34a (miR-34a) is a short noncoding regulatory RNA with tumor suppressive attributes. We leveraged our unique, large, population-based prognostic study of bladder cancer in New Hampshire, United States to evaluate miR-34a expression levels in individual tumor cells to assess prognostic value. We collected detailed exposure and medical history data, as well as tumor tissue specimens from bladder patients and followed them long-term for recurrence, progression and survival. Fluorescence-based in situ hybridization assays were performed on urothelial carcinoma tissue specimens (n\u2009=\u2009229). A larger proportion of the nonmuscle invasive tumors had high levels of miR-34a within the carcinoma cells compared to those tumors that were muscle invasive. Patients with high miR-34a levels in their baseline nonmuscle invasive tumors experienced lower risks of recurrence (adjusted hazard ratio 0.57, 95% confidence interval 0.34-0.93). Consistent with these observations, we demonstrated a functional tumor suppressive role for miR-34a in cultured urothelial cells, including reduced matrigel invasion and growth in soft agar. Our results highlight the need for further clinical studies of miR-34a as a guide for recurrence screening and as a possible candidate therapeutic target in the bladder.LM009012/LM/NLM NIH HHSUnited States/R01 CA057494/CA/NCI NIH HHSUnited States/P20 GM103534/GM/NIGMS NIH HHSUnited States/P20 GM103506/GM/NIGMS NIH HHSUnited States/R21 CA141017/CA/NCI NIH HHSUnited States/ES00002/ES/NIEHS NIH HHSUnited States/ES07373/ES/NIEHS NIH HHSUnited States/CA82354/CA/NCI NIH HHSUnited States/5P42ES05947/ES/NIEHS NIH HHSUnited States/CA121382/CA/NCI NIH HHSUnited States/P30 ES000002/ES/NIEHS NIH HHSUnited States/P42 ES007373/ES/NIEHS NIH HHSUnited States/U58 DP000798/DP/NCCDPHP CDC HHSUnited States/P20 RR018787/RR/NCRR NIH HHSUnited States/R03 CA099500/CA/NCI NIH HHSUnited States/R01 CA078609/CA/NCI NIH HHSUnited States/RR028309/RR/NCRR NIH HHSUnited States/CA102327/CA/NCI NIH HHSUnited States/R03 CA121382/CA/NCI NIH HHSUnited States/CA078609/CA/NCI NIH HHSUnited States/P30 CA023108/CA/NCI NIH HHSUnited States/RR018787/RR/NCRR NIH HHSUnited States/K07CA102327/CA/NCI NIH HHSUnited States/P42 ES005947/ES/NIEHS NIH HHSUnited States/K07 CA102327/CA/NCI NIH HHSUnited States/CA141017/CA/NCI NIH HHSUnited States/U58/DP000798/DP/NCCDPHP CDC HHSUnited States/R21 CA182659/CA/NCI NIH HHSUnited States/CA182659/CA/NCI NIH HHSUnited States/GM103506/GM/NIGMS NIH HHSUnited States/R01 LM009012/LM/NLM NIH HHSUnited States/GM103534/GM/NIGMS NIH HHSUnited States/CA099500/CA/NCI NIH HHSUnited States/RR024475/RR/NCRR NIH HHSUnited States/R01 CA082354/CA/NCI NIH HHSUnited States/P20 RR024475/RR/NCRR NIH HHSUnited States
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