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MiR-136 modulates glioma cell sensitivity to temozolomide by targeting astrocyte elevated gene-1.

Wu H, Liu Q, Cai T, Chen YD, Liao F, Wang ZF - Diagn Pathol (2014)

Bottom Line: Luciferase assays revealed that miR-136 directly targeted the 3'UTR of AEG-1. qRT-PCR and western blotting analysis found that AEG-1 expression at the mRNA and protein levels decreased in the miR-136 mimic-treatment group relative to control group.Downregulation of AEG-1 expression by siRNAs, U251 cells became more sensitive to the therapy of TMZ.In addition, the enhanced growth-inhibitory effect by the miR-136 mimics transfection was enhanced after the addition of AEG-1 siRNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, The Third Xiangya Hospital, Central South University, 138 Tongzipo Road, Hexi Yuelu District, Changsha 400013, Hunan, China. dr_wangzhifei@126.com.

ABSTRACT

Background: Recent studies have linked chemotherapy resistance to the altered expression of microRNAs (miRNAs). Thus, miRNA-based approaches to modulating sensitivity to temozolomide (TMZ) may overcome chemoresistance. The aim of the present study was to investigate whether miR-136 could modulates glioma cell sensitivity to TMZ.

Methods: The proliferation of glioma U251 cell line was evaluated by MTT assay. The expression of astrocyte elevated gene-1 (AEG-1)was detected by real‑time polymerase chain reaction (RT-PCR)and Western blot. The luciferase reporter gene was used to test whether AEG-1 was the target of the miR-136.

Results: The MTT assay showed that U251 cells with miR-136 overexpression were significantly more sensitive to the therapy of TMZ than control cells. Luciferase assays revealed that miR-136 directly targeted the 3'UTR of AEG-1. qRT-PCR and western blotting analysis found that AEG-1 expression at the mRNA and protein levels decreased in the miR-136 mimic-treatment group relative to control group. Downregulation of AEG-1 expression by siRNAs, U251 cells became more sensitive to the therapy of TMZ. In addition, the enhanced growth-inhibitory effect by the miR-136 mimics transfection was enhanced after the addition of AEG-1 siRNA.

Conclusions: The present study provides the first evidence that miR-136 plays a key role in TMZ resistance by targeting the AEG-1 protein in glioma cell line, suggesting that miR-136 can be used to predict a patient's response to TMZ therapy as well as serve as a novel potential maker for glioma therapy.

Virtual slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_173.

No MeSH data available.


Related in: MedlinePlus

Effect of the overexpression of miR‑136 on the cytotoxic activity of TMZ in U251 cells. (A) miR‑136‑transfected cells showed higher miR‑136 expression than the Con (P < 0.05) and miR-Con (P < 0.05) groups. (B) Cell viability was detected by MTT assay in pre-miR-136 transfected cells and Con and miR-Con groups following treatment with TMZ for 24 h.*P < 0.05.
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Fig1: Effect of the overexpression of miR‑136 on the cytotoxic activity of TMZ in U251 cells. (A) miR‑136‑transfected cells showed higher miR‑136 expression than the Con (P < 0.05) and miR-Con (P < 0.05) groups. (B) Cell viability was detected by MTT assay in pre-miR-136 transfected cells and Con and miR-Con groups following treatment with TMZ for 24 h.*P < 0.05.

Mentions: In order to explore the role of miR-136 in U251 cells, transfection with plasmids, pre-miR-136 or scrambled pre-miR control, was performed. The expression Levels of miR −136 were confirmed by qPCR, as shown in Figure 1A. Pre- miR-136 (miR −136) - transfected cells showed a higher miR-136 expression than the untransfected negative control (Con) and empty vector-transfected (miR-Con) groups. To evaluate the effect of miR-136 on the cytotoxic activity of TMZ in U251 cells, MTT assay was performed on the pre- miR-136 transfected cells, the Con and miR-Con groups combined with various concentrations of TMZ. The results showed that the viability of U251 cells with miR-136 overexpression was significantly decreased compared with that of the miR-Con group at the same concentration ofFigure 1


MiR-136 modulates glioma cell sensitivity to temozolomide by targeting astrocyte elevated gene-1.

Wu H, Liu Q, Cai T, Chen YD, Liao F, Wang ZF - Diagn Pathol (2014)

Effect of the overexpression of miR‑136 on the cytotoxic activity of TMZ in U251 cells. (A) miR‑136‑transfected cells showed higher miR‑136 expression than the Con (P < 0.05) and miR-Con (P < 0.05) groups. (B) Cell viability was detected by MTT assay in pre-miR-136 transfected cells and Con and miR-Con groups following treatment with TMZ for 24 h.*P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4195982&req=5

Fig1: Effect of the overexpression of miR‑136 on the cytotoxic activity of TMZ in U251 cells. (A) miR‑136‑transfected cells showed higher miR‑136 expression than the Con (P < 0.05) and miR-Con (P < 0.05) groups. (B) Cell viability was detected by MTT assay in pre-miR-136 transfected cells and Con and miR-Con groups following treatment with TMZ for 24 h.*P < 0.05.
Mentions: In order to explore the role of miR-136 in U251 cells, transfection with plasmids, pre-miR-136 or scrambled pre-miR control, was performed. The expression Levels of miR −136 were confirmed by qPCR, as shown in Figure 1A. Pre- miR-136 (miR −136) - transfected cells showed a higher miR-136 expression than the untransfected negative control (Con) and empty vector-transfected (miR-Con) groups. To evaluate the effect of miR-136 on the cytotoxic activity of TMZ in U251 cells, MTT assay was performed on the pre- miR-136 transfected cells, the Con and miR-Con groups combined with various concentrations of TMZ. The results showed that the viability of U251 cells with miR-136 overexpression was significantly decreased compared with that of the miR-Con group at the same concentration ofFigure 1

Bottom Line: Luciferase assays revealed that miR-136 directly targeted the 3'UTR of AEG-1. qRT-PCR and western blotting analysis found that AEG-1 expression at the mRNA and protein levels decreased in the miR-136 mimic-treatment group relative to control group.Downregulation of AEG-1 expression by siRNAs, U251 cells became more sensitive to the therapy of TMZ.In addition, the enhanced growth-inhibitory effect by the miR-136 mimics transfection was enhanced after the addition of AEG-1 siRNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, The Third Xiangya Hospital, Central South University, 138 Tongzipo Road, Hexi Yuelu District, Changsha 400013, Hunan, China. dr_wangzhifei@126.com.

ABSTRACT

Background: Recent studies have linked chemotherapy resistance to the altered expression of microRNAs (miRNAs). Thus, miRNA-based approaches to modulating sensitivity to temozolomide (TMZ) may overcome chemoresistance. The aim of the present study was to investigate whether miR-136 could modulates glioma cell sensitivity to TMZ.

Methods: The proliferation of glioma U251 cell line was evaluated by MTT assay. The expression of astrocyte elevated gene-1 (AEG-1)was detected by real‑time polymerase chain reaction (RT-PCR)and Western blot. The luciferase reporter gene was used to test whether AEG-1 was the target of the miR-136.

Results: The MTT assay showed that U251 cells with miR-136 overexpression were significantly more sensitive to the therapy of TMZ than control cells. Luciferase assays revealed that miR-136 directly targeted the 3'UTR of AEG-1. qRT-PCR and western blotting analysis found that AEG-1 expression at the mRNA and protein levels decreased in the miR-136 mimic-treatment group relative to control group. Downregulation of AEG-1 expression by siRNAs, U251 cells became more sensitive to the therapy of TMZ. In addition, the enhanced growth-inhibitory effect by the miR-136 mimics transfection was enhanced after the addition of AEG-1 siRNA.

Conclusions: The present study provides the first evidence that miR-136 plays a key role in TMZ resistance by targeting the AEG-1 protein in glioma cell line, suggesting that miR-136 can be used to predict a patient's response to TMZ therapy as well as serve as a novel potential maker for glioma therapy.

Virtual slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_173.

No MeSH data available.


Related in: MedlinePlus