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MiR-192 directly binds and regulates Dicer1 expression in neuroblastoma.

Feinberg-Gorenshtein G, Guedj A, Shichrur K, Jeison M, Luria D, Kodman Y, Ash S, Feinmesser M, Edry L, Shomron N, Weizman A, Yaniv I, Avigad S - PLoS ONE (2013)

Bottom Line: We were able to show through a dual luciferase assay and side-directed mutational analysis that miR-192 directly binds the 3' UTR of Dicer1 on positions 1232-1238 and 2282-2288.An increase in cell viability, proliferation and migration rates were evident in NB cells transfected with miR-192-mimic.Yet, there was a significant decrease in proliferation when NB cells were transfected with an miR-192-inhibitor We suggest that miR-192 might be a key player in NB by regulating Dicer1 expression.

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology, Felsenstein Medical Research Center, Petah Tikva, Israel ; Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

ABSTRACT
Neuroblastoma (NB) arises from the embryonic neural crest and is the most common extracranial solid tumor in children under 5 years of age. Reduced expression of Dicer1 has recently been shown to be in correlation with poor prognosis in NB patients. This study aimed to investigate the mechanisms that could lead to the down-regulation of Dicer1 in neuroblastoma. We used computational prediction to identify potential miRs down-regulating Dicer1 in neuroblastoma. One of the miRs that were predicted to target Dicer1 was miR-192. We measured the levels of miR-192 in 43 primary tumors using real time PCR. Following the silencing of miR-192, the levels of dicer1 cell viability, cell proliferation and migration capability were analyzed. Multivariate analysis identified miR-192 as an independent prognostic marker for relapse in neuroblastoma patients (p=0.04). We were able to show through a dual luciferase assay and side-directed mutational analysis that miR-192 directly binds the 3' UTR of Dicer1 on positions 1232-1238 and 2282-2288. An increase in cell viability, proliferation and migration rates were evident in NB cells transfected with miR-192-mimic. Yet, there was a significant decrease in proliferation when NB cells were transfected with an miR-192-inhibitor We suggest that miR-192 might be a key player in NB by regulating Dicer1 expression.

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Related in: MedlinePlus

miR-192 is associated with NB cell viability, proliferation and migration capability.Following transfection of miR-192 mimic and inhibitor, we measured the viability of NUB6 cells by direct counting (A), proliferation properties of the cells by XTT analysis (B) and the migration ability of NUB6 cells (C).A. Inhibition of miR-192 resulted in significantly decreased cell viability (*p=0.042).B. Following transfection of miR-192 mimic and inhibitor, a significant increase(*p=0.044) or decrease (*p=0.038) was detected in the proliferation rate respectively.C. Overexpressing miR-192 resulted in a significant increase in cell migration (*p= 0.00036). Values are expressed as the mean ± SD of combined results of three independent experiments.
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pone-0078713-g007: miR-192 is associated with NB cell viability, proliferation and migration capability.Following transfection of miR-192 mimic and inhibitor, we measured the viability of NUB6 cells by direct counting (A), proliferation properties of the cells by XTT analysis (B) and the migration ability of NUB6 cells (C).A. Inhibition of miR-192 resulted in significantly decreased cell viability (*p=0.042).B. Following transfection of miR-192 mimic and inhibitor, a significant increase(*p=0.044) or decrease (*p=0.038) was detected in the proliferation rate respectively.C. Overexpressing miR-192 resulted in a significant increase in cell migration (*p= 0.00036). Values are expressed as the mean ± SD of combined results of three independent experiments.

Mentions: Next, we were interested in investigating the affect of miR-192 on the aggressiveness of NB cells. NUB6 cells transfected with miR-192-inhibitor led to a 39% decrease in cell viability (p=0.042, Figure 7A). NUB6 cells transfected with miR-192-mimic and inhibitor led to an increase of 17% in proliferation (p=0.044) or a significant decrease of 16% in the proliferation rate (p=0.038, Figure 7B), respectively. Over-expressing miR-192 exhibited a significantly increased migration ability (a 1.7-fold increase, p= 0.00036) as compared to the negative control (Figure 7C).


MiR-192 directly binds and regulates Dicer1 expression in neuroblastoma.

Feinberg-Gorenshtein G, Guedj A, Shichrur K, Jeison M, Luria D, Kodman Y, Ash S, Feinmesser M, Edry L, Shomron N, Weizman A, Yaniv I, Avigad S - PLoS ONE (2013)

miR-192 is associated with NB cell viability, proliferation and migration capability.Following transfection of miR-192 mimic and inhibitor, we measured the viability of NUB6 cells by direct counting (A), proliferation properties of the cells by XTT analysis (B) and the migration ability of NUB6 cells (C).A. Inhibition of miR-192 resulted in significantly decreased cell viability (*p=0.042).B. Following transfection of miR-192 mimic and inhibitor, a significant increase(*p=0.044) or decrease (*p=0.038) was detected in the proliferation rate respectively.C. Overexpressing miR-192 resulted in a significant increase in cell migration (*p= 0.00036). Values are expressed as the mean ± SD of combined results of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815303&req=5

pone-0078713-g007: miR-192 is associated with NB cell viability, proliferation and migration capability.Following transfection of miR-192 mimic and inhibitor, we measured the viability of NUB6 cells by direct counting (A), proliferation properties of the cells by XTT analysis (B) and the migration ability of NUB6 cells (C).A. Inhibition of miR-192 resulted in significantly decreased cell viability (*p=0.042).B. Following transfection of miR-192 mimic and inhibitor, a significant increase(*p=0.044) or decrease (*p=0.038) was detected in the proliferation rate respectively.C. Overexpressing miR-192 resulted in a significant increase in cell migration (*p= 0.00036). Values are expressed as the mean ± SD of combined results of three independent experiments.
Mentions: Next, we were interested in investigating the affect of miR-192 on the aggressiveness of NB cells. NUB6 cells transfected with miR-192-inhibitor led to a 39% decrease in cell viability (p=0.042, Figure 7A). NUB6 cells transfected with miR-192-mimic and inhibitor led to an increase of 17% in proliferation (p=0.044) or a significant decrease of 16% in the proliferation rate (p=0.038, Figure 7B), respectively. Over-expressing miR-192 exhibited a significantly increased migration ability (a 1.7-fold increase, p= 0.00036) as compared to the negative control (Figure 7C).

Bottom Line: We were able to show through a dual luciferase assay and side-directed mutational analysis that miR-192 directly binds the 3' UTR of Dicer1 on positions 1232-1238 and 2282-2288.An increase in cell viability, proliferation and migration rates were evident in NB cells transfected with miR-192-mimic.Yet, there was a significant decrease in proliferation when NB cells were transfected with an miR-192-inhibitor We suggest that miR-192 might be a key player in NB by regulating Dicer1 expression.

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology, Felsenstein Medical Research Center, Petah Tikva, Israel ; Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

ABSTRACT
Neuroblastoma (NB) arises from the embryonic neural crest and is the most common extracranial solid tumor in children under 5 years of age. Reduced expression of Dicer1 has recently been shown to be in correlation with poor prognosis in NB patients. This study aimed to investigate the mechanisms that could lead to the down-regulation of Dicer1 in neuroblastoma. We used computational prediction to identify potential miRs down-regulating Dicer1 in neuroblastoma. One of the miRs that were predicted to target Dicer1 was miR-192. We measured the levels of miR-192 in 43 primary tumors using real time PCR. Following the silencing of miR-192, the levels of dicer1 cell viability, cell proliferation and migration capability were analyzed. Multivariate analysis identified miR-192 as an independent prognostic marker for relapse in neuroblastoma patients (p=0.04). We were able to show through a dual luciferase assay and side-directed mutational analysis that miR-192 directly binds the 3' UTR of Dicer1 on positions 1232-1238 and 2282-2288. An increase in cell viability, proliferation and migration rates were evident in NB cells transfected with miR-192-mimic. Yet, there was a significant decrease in proliferation when NB cells were transfected with an miR-192-inhibitor We suggest that miR-192 might be a key player in NB by regulating Dicer1 expression.

Show MeSH
Related in: MedlinePlus