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15 publications mentioning hsa-mir-544a

Open access articles that are associated with the species Homo sapiens and mention the gene name mir-544a. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1
[+] score: 156
Under these experimental conditions, the 3’ UTR SELK segment 269–290, which is supposed to be targeted by miR-544a, inhibited significantly the expression of the reporter (Fig 1A); in particular, the activity registered for the reporter construct was found 26% lower than that obtained with the parental vector, indicating a direct interaction between miR-544a and the SELK sequence. [score:8]
This boost resulted in an additional luciferase reduction on the reporter construct WT (about 30%) in comparison to the inhibition due to the endogenously expressed miR-544a (see the results obtained by transfecting WT construct together with the negative control mimc, Ctrl-miRNA), leading to a total inhibition equal to 60% (Fig 3A). [score:7]
0156908.g005 Fig 5 Effect of sodium selenite on miR-544a expression and SELK (A) Cells were cultured for 24 h and 48 h in standard condition or in presence of increasing sodium selenite doses; miR-544a expression was determined by RT-qPCR and reported as fold change of expression in treated samples vs untreated ones. [score:7]
In particular, Mo et al. (2014) [37] reported that miR-544a down-regulates CDH1 (E-cadherin) and up-regulates vimentin in non-small cell lung cancer (NSCLC). [score:7]
In particular, it is very important to underline that, as shown in Fig 5, after the treatment of HCC cell lines with sodium selenite, the gene expression trend of SELK was towards increasing, while those of miR-544a towards a decrease, thus suggesting that the selenium is able to block the ability of miR-544a to negatively regulate the SELK expression. [score:6]
Overall, our data evidenced that: i) only miR-544a was able to silence, at physiological concentration, the reporter gene by interacting with the 3’ UTR SELK segment 269–290 (Fig 1A), ii) the observed silencing effect was specifically due to miR-544a, because it was modulated by miR-544a mimic and inhibitor (Fig 1B), iii) miR-544a is able to interfere with SELK translation through a naturally occurring interaction in hepatic cells (Figs 3 and 4). [score:5]
0156908.g003 Fig 3 HuH-7 and HepG2 cells were transfected with the luciferase -based reporter plasmid psiCheck-2 containing the SELK target sequence for miR-544a (WT) or a control DNA with inverted sequence (I), along with 50 nM miR-544a mimic (miR-544a, A and B) or 50 nM miR-544a inhibitor (anti-miR-544a, C and D). [score:5]
HepG2 and HuH-7 cells were transfected with 0.2 and 0.05 μg of reporter constructs, respectively; miScript miR-544a mimic, miScript miR-544a inhibitor, and their controls with unrelated sequences (AllStars Negative Control and miScript Inhibitor Negative Control, respectively; all from Qiagen) were transfected at 50nM along with 50ng of phRL-tk plasmid encoding Renilla luciferase to monitor transfection efficiency. [score:5]
This screening of target validation revealed that miR-544a was able to silence the SELK expression in HCC cell lines. [score:5]
48h after transfection, luciferase activities were recorded; the Renilla luciferase activity (Rl) was normalized to the firefly luciferase activity (luc) and the uninhibited activity relative to the parental vector psiCheck2 (V) was set to 1. (B) Expression of miRNAs were evaluated by qRT-PCR and reported as arbitrary units (A. U. ) (C) Following the same procedure as above (A), a control construct (I), containing the inverted sequence targeted by miR-544a, was tested. [score:5]
HuH-7 and HepG2 cells were transfected with the luciferase -based reporter plasmid psiCheck-2 containing the SELK target sequence for miR-544a (WT) or a control DNA with inverted sequence (I), along with 50 nM miR-544a mimic (miR-544a, A and B) or 50 nM miR-544a inhibitor (anti-miR-544a, C and D). [score:5]
In contrast, miR-181a and miR-181b had higher expression level than miR-544a and target the same reporter construct; however, no silencing effect was observed. [score:5]
In particular, 24 h after treatment, even at 0.25 μM sodium selenite exerts a repressive effect on the expression of miR-544a (approximately 3.5-fold inhibition), when compared to untreated samples. [score:4]
These results clearly indicate that miR-544a is able to interfere with the SELK translation providing direct evidence that the interaction between the selected miRNA and SELK occurs naturally in hepatic cells. [score:4]
These experiments were also performed in HepG2 cell line (Fig 3C and 3D) to generalize the results, also considering the comparable expression levels of miR-544a in HuH-7 and HepG2 cells (Fig 2), and to verify that this miRNA is not influenced from wild-type or mutated p53. [score:3]
Representative of protein extracts at 48 h after transfection with miR-544a mimic (A) or miR-544a inhibitor (B) and their relative control molecules at 50 nM. [score:3]
Western blot analyses revealed a reduced amount of SELK as a consequence of miR-544a mimic transfection and an increase of SELK after miR-544a inhibitor transfection (Fig 4). [score:3]
Furthermore, the role of miR-544a was tested also in a hypoxic breast cancer mo del because it was able to silence the mammalian target of rapamycin (mTOR) [39]. [score:3]
Small Molecule Inhibition of miR-544 Biogenesis Disrupts Adaptive Responses to Hypoxia by Modulating ATM-mTOR Signaling. [score:3]
The(se) same Authors have also shown that miR-544a, while decreasing the CDH1 and AXIN2 expression, at the same time induces the nuclear import of β-catenin and stabilizes β-catenin in the nucleus by means of Wnt signaling pathway activation [34]. [score:3]
In particular, these studies could also highlight whether miR-544a and/or SELK expression level might be useful for improving the diagnosis and/or prognosis of this cancer. [score:3]
Overall, the screening of the SELK target sequences indicates that at physiological concentration miR-544a is able to silence the reporter gene by interacting with the 3’ UTR SELK segment 269–290. [score:3]
miR-544a expression in three common HCC cell lines. [score:3]
Expression of miR-544a in HCC cell lines. [score:3]
0156908.g004 Fig 4Representative of protein extracts at 48 h after transfection with miR-544a mimic (A) or miR-544a inhibitor (B) and their relative control molecules at 50 nM. [score:3]
Hence, we focused on miR-544a, demonstrating that it interferes with SELK translation and is modulated by selenium treatment. [score:3]
Hence, we can highlight that the silencing effect observed for miR-544a construct depend on its ability to interact with SELK sequence and was strong, taking into account the relative expression levels of the different miRNAs. [score:3]
Control plasmid (indicated as I) for miR-544a was obtained by the same approach, i. e. cloning the couple of oligonucleotides representing the inverted target sequence (5’-TAAGACGTTGGATTTTTATAAA-3’) in psiCheck-2 vector. [score:3]
As shown in Fig 2, miR-544a shows a comparable level of expression in HuH-7 and HepG2 cells, and a lower extent in PLC/PRF/5. [score:3]
0156908.g002 Fig 2 miR-544a expression levels were evaluated by RT-qPCR and reported as fold change relative to that of HuH-7. The silencing effect of miR-544a was then investigated by transfecting both reporter constructs (I and WT) along with miR-544a mimic and miR-544a inhibitor, as well as their negative control molecules, Ctrl-miRNA and Ctrl-anti-miRNA, which do not match the target sequence. [score:3]
0156908.g002 Fig 2 miR-544a expression levels were evaluated by RT-qPCR and reported as fold change relative to that of HuH-7. miR-544a expression levels were evaluated by RT-qPCR and reported as fold change relative to that of HuH-7. The silencing effect of miR-544a was then investigated by transfecting both reporter constructs (I and WT) along with miR-544a mimic and miR-544a inhibitor, as well as their negative control molecules, Ctrl-miRNA and Ctrl-anti-miRNA, which do not match the target sequence. [score:3]
No difference was observed on the proliferation of HuH-7 and HepG2 cell lines transfected with miR-544a mimic and inhibitor compared to their relative control molecules, as already reported for other cell lines (see S1 Fig) [34]. [score:2]
Furthermore, since SELK was found to be induced by sodium selenite [26], we have also investigated whether miR-544a was responsive to selenium, finding that both miR-544a and SELK expression were conversely regulated by sodium selenite. [score:2]
Conversely, the inhibition of miR-544a activity resulted in a recovery of luciferase activity when it was compared to that measured by transfecting the WT construct together with the negative control inhibitor (Ctrl-anti-miRNA) (Fig 3B). [score:2]
The assays described above are based on the silencing effect of miR-544a on the translation of a chimeric mRNA containing the Renilla luciferase coding sequence and a segment of SELK UTR mRNA. [score:2]
Real-time PCR analyses revealed that miR-544a was sensitive to the selenium (Fig 5A). [score:1]
This point was addressed by transfecting cells with miR-544a mimic, or inhibitor, and evaluating their effect on the SELK protein. [score:1]
In general, the biological function(s) of miR-544a is still largely unknown, since scarce data are reported in the literature. [score:1]
S1 Fig HuH-7 cells were transfected with miR-544a mimic or anti-miR-544a and their relative control molecules at 50 nM. [score:1]
Overall, these results suggest that miR-544a could be in part responsible of SELK induction by sodium selenite. [score:1]
MiR-544a mimic increased the level of endogenous miR-544a of approximately 300-fold, as determined by RT-QPCR (data not shown). [score:1]
Moreover, on the basis of data related to the treatment of HCC cells with sodium selenite, we can suggest an intriguing network, among between selenium, as essential micronutrient, a miRNA, i. e., miR-544a, and a protein, i. e., SELK. [score:1]
We investigated whether miR-544a expression was modulated by selenium and what was the selenium role in the induction of SELK. [score:1]
The data clearly indicate that the silencing effect of cellular miRNAs toward the 3’ UTR SELK segment 269–290 can be attributed to miR-544a in both tested HCC cell lines. [score:1]
HuH-7 cells were transfected with miR-544a mimic or anti-miR-544a and their relative control molecules at 50 nM. [score:1]
Interfering activity of miR-544a in HCC cell lines. [score:1]
Since no literature data are known about miR-544a involvement in HCC, we evaluated also its expression levels in some common HCC cell lines. [score:1]
As a further control, the reporter activity was also registered after transfection of a construct containing the inverted sequence recognized by miR-544a (I); the luciferase activity was found to be comparable to that determined after transfection of the parental vector and significantly higher than that obtained with the wild-type construct (WT) (Fig 1C), as expected. [score:1]
However, in this context it is worth of note to underline that in all these studies, the miR-544a was not reported, and, hence, our results indicates for the first time its involvement in HCC. [score:1]
Effect of sodium selenite on miR-544a interfering activity. [score:1]
Silencing activity of hsa-miR-544a. [score:1]
Effect on cell proliferation of miR-544a. [score:1]
The silencing effect of miR-544a on SELK in HuH-7 cells. [score:1]
In details, having investigated the silencing effect of miR-544a in HuH-7 cells, we compared the expression of this miRNA with other two HCC cell lines showing a different phenotype with respect to HuH-7 cells with no viral infection but p53 mutation (mutant p53 Y220C) [30]. [score:1]
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2
[+] score: 95
Other miRNAs from this paper: hsa-mir-107, hsa-mir-34a, hsa-mir-140, hsa-mir-143, hsa-mir-874
Western blot analysis revealed that in cells stably expressing miR-544a, the level of GSK3β was reduced, whereas the expression levels of β-catenin and CD133 were upregulated. [score:8]
The miR-544a mimic could not interact with and inhibit the expression of the GSK3β MUT 3′UTR, and the expression of reporter gene (1.01±0.02) increased (q=491.05, P<0.01). [score:7]
The effects of miR-544a were studied in 95C and 95D cells in order to reveal the mechanism of GSK3β downregulation, an inhibitory factor of the Wnt pathway (7). [score:6]
A retroviral vector pBaBe-puro (Addgene, Cambridge, MA, USA) expressing miR-544a was constructed and then inoculated into HEK293T cells for 24 h. The reagents were added to a 1.5 ml Eppendorf tube, including 20 μg PIK, 20 μg expression plasmid, 110 μl ddH [2]O, 250 μl CaCl and 200 μl hepes-buffered saline. [score:5]
validated that miR-544a could interact with and inhibit the expression of GSK3β. [score:5]
Identification of cells stably expressing miR-544a, byThe expression level of 95C NC and 95D NC was normalized to 1.00±0.00. [score:5]
Luciferase expression was given as the relative light units (RLUC/LUC) to determine whether GSK3β was the target of miR-544a in vitro. [score:5]
The miR-544a target gene, GSK3β, was predicted using TargetScan software (http://www. [score:5]
The relative fold change of expression of miR-544a was quantified as 2 [−ΔΔ]Ct, where ΔΔCt was Ct (target gene) - Ct (housekeeping gene). [score:5]
Bioinformatic analyses indicated that miR-544a targeted GSK3β, an inhibitory factor of the Wnt pathway. [score:5]
As shown in Table I, luciferase assays revealed that the miR-544a mimic can interact and inhibit the expression of GSK3β 3′UTR (0.52±0.01). [score:4]
Abnormal expression of miR-544a leads to NSCLC self-renewal. [score:3]
Identification of cells stably expressing miR-544a, by qPCR. [score:3]
showed that cells stably expressing miR-544a (95C+miR-544a or 95D+miR-544a) had an increased tendency to form tumor spheroids (Fig. 3). [score:3]
Protein expression was quantitatively assessed using an HRP-enhanced chemiluminesence scanner (LAS-4000 mini luminescent imaging analyzer; Fijifilm, Tokyo, Japan) 95C, 95D, miR-544a-95C and miR-544a-95D cells were digested with 0.25% pancreatic enzyme, and 1,000 cells/ml were resuspended in RPMI-1640 serum-free medium. [score:3]
Spheroid culture showed that cells stably expressing miR-544a (95C+miR-544a or 95D+miR-544a) had an increased tendency to form tumor spheroids (Fig. 3). [score:3]
It was observed that the cells stably expressing miR-544a had an increased tendency to form tumor CD133 -positive spheroids. [score:3]
Following transfection with miR-544a, the miR-544a expression level of 95C and 95D cells was 20.51±0.97 and 15.16±1.38, respectively (F=418.05, P<0.01), among all four groups. [score:3]
Validation of miR-544a target gene by luciferase assay. [score:2]
The results showed that miR-544a was highly likely to interact with GSK3β (Fig. 1). [score:1]
In conclusion, miR-544a has an important function not only in tumor invasion and metastasis, but also in TSC formation. [score:1]
According to the western blot analysis, the level of GSK3β reduced, but that of β-catenin and CD133 increased in 95C and 95D cells transfected with miR-544a. [score:1]
To determine the impact of miR-544a in spheroid formation, a spheroid culture was established. [score:1]
Total RNA was extracted from 95C, 95D, miR-544a-95C and miR-544a-95D cells using TRIzol™ reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) and reverse-transcribed to cDNA using M-MLV reverse transcriptase (Toyobo Co. [score:1]
It was therefore concluded that miR-544a activated the Wnt pathway (Fig. 2 and Table III). [score:1]
The present study aimed to determine the function of miR-544a in the formation of TSCs. [score:1]
miRNA negative control (NC) and miR-544a mimic (MC) were transfected together with GSK3β 3′ UTR or GSK3β mutated (MUT) 3′UTR, respectively, into HEK293T cells obtained from the American Type Culture Collection (Manassa, VA, USA) for 24 h according to the manufacturer’s instructions (Promega Corporation). [score:1]
Future studies will focus on the mechanism of miR-544a in the formation of TSCs, with a view to novel NSCLC treatment approaches. [score:1]
miR-544a has been shown to promote tumor invasion and metastasis (11). [score:1]
95C and 95D cells (American Type Culture Collection) were subsequently infected by these viruses and the cells with highest levels of miR-544a were screened using a puromycin marker. [score:1]
Effect of miR-544a on spheroid formation. [score:1]
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3
[+] score: 31
Other miRNAs from this paper: hsa-mir-134, hsa-mir-154, hsa-mir-379, hsa-mir-382, hsa-mir-544b
We conclude that downregulation of 14q32 miRNA expression is an evolutionarily conserved mechanism that contributes to the biological behavior of osteosarcoma, and that quantification of representative transcripts from this family, such as miR-382, miR-134, and miR-544, provide prognostic and predictive markers that can assist in the management of patients with this disease. [score:8]
We also show a comparable decrease in expression of orthologous 14q32 miRNAs in canine osteosarcoma samples, with conservation of the inverse correlation between aggressive behavior and expression of orthologous miRNA miR-134 and miR-544. [score:5]
Group 1 was composed of tumor samples with miR-134 (Figure  4A) and miR-544 (Figure  4B) expression levels above the median survival days and Group 2 was composed of tumor samples below the median, and these two groups were interrogated via Kaplan-Meier survival analysis (Figure  4D). [score:3]
Dogs with lowest levels of miR-134 or miR-544 expression showed decreased likelihood of survival. [score:3]
Similar results were obtained using miR-544 expression levels (p-value 0.012, Figure  4D). [score:3]
Two miRNAs (miR-134 and miR-544) that showed 100% conservation and mapped to the predicted region of synteny in canine chromosome (CFA) 8 were used to examine expression of the 14q32 cluster in dog samples (Additional file 5: Figure S1). [score:3]
As observed in human osteosarcoma the levels of miR-544 and miR-134 were directly correlated (R [2] = 0.88) Figure 4 14q32 orthologous region transcript levels and outcome in canine osteosarcoma. [score:2]
As observed in human osteosarcoma the levels of miR-544 and miR-134 were directly correlated (R [2] = 0.88) Figure 4 14q32 orthologous region transcript levels and outcome in canine osteosarcoma. [score:2]
Survival of dogs with osteosarcoma in Group 1 and Group 2 are significantly different (miR-134 p-value 0.004, miR-544 p-value 0.01). [score:1]
To address this, we determined the levels of miR-544 and miR-134 (based on identical mature miRNA sequences) in 16 canine osteosarcoma tumors from which RNA was available by (Figure  4A). [score:1]
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4
[+] score: 11
Of the 209 differentially expressed genes and LncRNA, 73 were upregulated (right, red) and 136 were downregulated (left, green) in stage 1 compared with stage 0. (a) Alignment between XLOC005698 and oar-miR-3955-5p as well as between XLOC000629 and oar-miR-544-5p. [score:8]
0156890.g006 Fig 6 (a) Alignment between XLOC005698 and oar-miR-3955-5p as well as between XLOC000629 and oar-miR-544-5p. [score:1]
BLAST alignment analysis was performed for the differential lncRNAs with the mature miRNA of sheep from the miRBase database, and the results showed a high consistency between XLOC005698 and oar-miR-3955-5p as well as between XLOC000629 and oar-miR-544-5p (Fig 6A). [score:1]
Bioinformatics predicted oar-miR-3955-5p binding sites on XLOC005698, and oar-miR-544-5p binding sites on XLOC000629. [score:1]
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5
[+] score: 8
Other miRNAs from this paper: hsa-mir-185, hsa-mir-544b
For example, miRNA-544 directly targets the 3′-UTR of the newly-identified tumor suppressor gene IRX1, whose hypermethylation decreases expression of the protein in GC [40]. [score:8]
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6
[+] score: 7
org, we choose miR-186, miR-384, miR-410, miR-448, miR-496 and miR-544 to further study which miRNA might regulate matrilin-3 expression. [score:4]
a The mRNA and protein levels of matrilin-3 in normal and osteoarthritis cartilages were determined using quantitative real-time PCR and western blot, n = 10. b The levels of miR-186, miR-384, miR-410, miR-448, miR-496 and miR-544 in osteoarthritis cartilages were determined using quantitative real-time PCR, n = 10. c Pearson’s correlation analysis of the relative expression levels of miR-448 and the relative matrilin-3 mRNA levels in osteoarthritis cartilage. [score:3]
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7
[+] score: 7
[34] Mao et al. reported that miR-544 is involved in cell-cycle regulation and suppresses cervical cancer cell proliferation, colony formation, migration and invasion in a manner associated with YWHAZ downregulation in cervical cancer. [score:7]
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8
[+] score: 7
These included predicted binding of spliced genes to three forms of hsa-miR-376 (a–c forms), hsa-mir-544 targeted at the same spliced gene (ITGAL) and the inflammation-related hsa-mir-150 [87] predicted to target the disease spliced gene FGD4. [score:7]
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9
[+] score: 6
Furthermore, reduced expression of 14q32 miRNAs in human OS tumors and orthologous miR-134 and miR-544 in canine OS is associated with shorter survival, suggesting that dysregulation of the 14q32 miRNA cluster may represent a conserved mechanism contributing to the aggressive biological behavior of OS in both species. [score:4]
Consistent with findings in human OS, cross-species comparative analysis found decreased expression of miR-134 and miR-544 (orthologous to the human 14q32 miRNA cluster) in canine OS tumors compared to reactive canine osteoblasts [37]. [score:2]
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10
[+] score: 6
They showed a lower miR-134 and miR-544 expression in canine and human bone tumors in comparison to healthy tissues (Sarver et al., 2013). [score:3]
The miR-134 and miR-544 of the 14q32 cluster, showing 100% homology between both species, were used to examine the expression in canine samples. [score:3]
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11
[+] score: 4
We confirmed that subset of these 50 miRNAs (miR-382, miR-369-5p, miR-544 and miR-134) could target the 3′ UTR of cMYC transcript. [score:3]
Out of the 50 miRNAs, 2 miRNAs, miR-134 and mir-544, shared 100% conservation with the canine genome and mapped to the predicted synteny in canine osteosarcoma [14]. [score:1]
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12
[+] score: 3
The bioinformatics predictions identified a subset of these 50 miRNAs (miR-382, miR-369-5p, miR-544, and miR-134) that could potentially target the 3′UTR of the cMYC transcript (Thayanithy et al., 2012). [score:3]
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13
[+] score: 2
miR-544 could be a key regulator in switching cell cycles on or off in GC [41]. [score:2]
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14
[+] score: 2
Specifically, reintroduction of 14q32 -associated miR-544, miR-369, miR-382, and miR-134 reduced steady state levels of cMYC protein and induced apoptosis of Saos2 cells. [score:1]
Sequences 200 nucleotides upstream to the transcription start site of miR-654, miR-431, miR-127, miR-432, miR-411, miR-544, miR-369-3p, miR-382 and miR-134 were then amplified using qPCR from DNA present in the immune complexes. [score:1]
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15
[+] score: 1
There are several miRNAs encoded in 14q32 locus, including miR-655-3p, miR-127-5p, miR-369-3p, miR-544a. [score:1]
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