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MicroRNA‑20a promotes the proliferation and cell cycle of human osteosarcoma cells by suppressing early growth response 2 expression.

Zhuo W, Ge W, Meng G, Jia S, Zhou X, Liu J - Mol Med Rep (2015)

Bottom Line: It was determined that miR‑20a expression is markedly upregulated in OS tissues and cells compared with the matched adjacent normal tissues and h‑FOB human osteoblast cell lines.Data from luciferase reporter assays showed that miR‑20a directly binds to the 3'‑untranslated region (3'‑UTR) of EGR2 mRNA and represses expression at the transcriptional and translational levels.In conclusion, the data provide compelling evidence that miR‑20a functions as an onco‑miRNA, which is important in promoting cell proliferation in OS, and its oncogenic effect is mediated primarily through direct suppression of EGR2 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics and Traumatology, Institute of Orthopedics and Traumatology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shanxi 710000, P.R. China.

ABSTRACT
MicroRNAs (miRNAs) are crucial in cancer development. However, the underlying mechanisms of miRNAs in osteosarcoma (OS) remain largely uncharacterized. The present study investigated the role of miR‑20a in OS cell proliferation. It was determined that miR‑20a expression is markedly upregulated in OS tissues and cells compared with the matched adjacent normal tissues and h‑FOB human osteoblast cell lines. Ectopic expression of miR‑20a promoted the proliferation and anchorage‑independent growth of OS cells, whereas inhibition of miR‑20a reduced this effect. Bioinformatics analysis further revealed early growth response 2 (EGR2), as a potential target of miR‑20a. Data from luciferase reporter assays showed that miR‑20a directly binds to the 3'‑untranslated region (3'‑UTR) of EGR2 mRNA and represses expression at the transcriptional and translational levels. In functional assays, miR‑20a promoted OS cell proliferation and the cell cycle, which could be suppressed by an inhibitor of miR‑20a. In conclusion, the data provide compelling evidence that miR‑20a functions as an onco‑miRNA, which is important in promoting cell proliferation in OS, and its oncogenic effect is mediated primarily through direct suppression of EGR2 expression.

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Related in: MedlinePlus

Expression of miR-20a in human OS tissues and cell lines. (A) Relative miR-20a expression levels in 8 paired primary OS tissues and matched ANTs from the same patient were detected by PCR analysis. (B) RT-qPCR analysis of miR-20a expression in h-FOB human osteoblast cell lines and MG-63, Saos-2 and SW1353 OS cell lines. Experiments were repeated at least three times. Each bar represents the mean of three independent experiments. *P<0.05, compared with h-FOB. miR, microRNA; OS, osteosarcoma; T, primary OS tissues; ANT, adjacent normal tissues; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
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f1-mmr-12-04-4989: Expression of miR-20a in human OS tissues and cell lines. (A) Relative miR-20a expression levels in 8 paired primary OS tissues and matched ANTs from the same patient were detected by PCR analysis. (B) RT-qPCR analysis of miR-20a expression in h-FOB human osteoblast cell lines and MG-63, Saos-2 and SW1353 OS cell lines. Experiments were repeated at least three times. Each bar represents the mean of three independent experiments. *P<0.05, compared with h-FOB. miR, microRNA; OS, osteosarcoma; T, primary OS tissues; ANT, adjacent normal tissues; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Mentions: To investigate the role of miR-20a in OS development, the expression levels of miR-20a in OS tissues and OS cells were first evaluated by RT-qPCR (Fig. 1A). The results showed that the expression levels of miR-20a were upregulated in the OS tissues compared with in the matched ANTs, and all three tested OS cell lines had significantly upregulated miR-20a levels compared with the h-FOB human osteoblast cell lines (Fig. 1B). These results showed that miR-20a exhibited a crucial role in OS and suggest that miR-20a is significantly increased in OS and may serve as a prognostic marker for patients with OS.


MicroRNA‑20a promotes the proliferation and cell cycle of human osteosarcoma cells by suppressing early growth response 2 expression.

Zhuo W, Ge W, Meng G, Jia S, Zhou X, Liu J - Mol Med Rep (2015)

Expression of miR-20a in human OS tissues and cell lines. (A) Relative miR-20a expression levels in 8 paired primary OS tissues and matched ANTs from the same patient were detected by PCR analysis. (B) RT-qPCR analysis of miR-20a expression in h-FOB human osteoblast cell lines and MG-63, Saos-2 and SW1353 OS cell lines. Experiments were repeated at least three times. Each bar represents the mean of three independent experiments. *P<0.05, compared with h-FOB. miR, microRNA; OS, osteosarcoma; T, primary OS tissues; ANT, adjacent normal tissues; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581803&req=5

f1-mmr-12-04-4989: Expression of miR-20a in human OS tissues and cell lines. (A) Relative miR-20a expression levels in 8 paired primary OS tissues and matched ANTs from the same patient were detected by PCR analysis. (B) RT-qPCR analysis of miR-20a expression in h-FOB human osteoblast cell lines and MG-63, Saos-2 and SW1353 OS cell lines. Experiments were repeated at least three times. Each bar represents the mean of three independent experiments. *P<0.05, compared with h-FOB. miR, microRNA; OS, osteosarcoma; T, primary OS tissues; ANT, adjacent normal tissues; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
Mentions: To investigate the role of miR-20a in OS development, the expression levels of miR-20a in OS tissues and OS cells were first evaluated by RT-qPCR (Fig. 1A). The results showed that the expression levels of miR-20a were upregulated in the OS tissues compared with in the matched ANTs, and all three tested OS cell lines had significantly upregulated miR-20a levels compared with the h-FOB human osteoblast cell lines (Fig. 1B). These results showed that miR-20a exhibited a crucial role in OS and suggest that miR-20a is significantly increased in OS and may serve as a prognostic marker for patients with OS.

Bottom Line: It was determined that miR‑20a expression is markedly upregulated in OS tissues and cells compared with the matched adjacent normal tissues and h‑FOB human osteoblast cell lines.Data from luciferase reporter assays showed that miR‑20a directly binds to the 3'‑untranslated region (3'‑UTR) of EGR2 mRNA and represses expression at the transcriptional and translational levels.In conclusion, the data provide compelling evidence that miR‑20a functions as an onco‑miRNA, which is important in promoting cell proliferation in OS, and its oncogenic effect is mediated primarily through direct suppression of EGR2 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics and Traumatology, Institute of Orthopedics and Traumatology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shanxi 710000, P.R. China.

ABSTRACT
MicroRNAs (miRNAs) are crucial in cancer development. However, the underlying mechanisms of miRNAs in osteosarcoma (OS) remain largely uncharacterized. The present study investigated the role of miR‑20a in OS cell proliferation. It was determined that miR‑20a expression is markedly upregulated in OS tissues and cells compared with the matched adjacent normal tissues and h‑FOB human osteoblast cell lines. Ectopic expression of miR‑20a promoted the proliferation and anchorage‑independent growth of OS cells, whereas inhibition of miR‑20a reduced this effect. Bioinformatics analysis further revealed early growth response 2 (EGR2), as a potential target of miR‑20a. Data from luciferase reporter assays showed that miR‑20a directly binds to the 3'‑untranslated region (3'‑UTR) of EGR2 mRNA and represses expression at the transcriptional and translational levels. In functional assays, miR‑20a promoted OS cell proliferation and the cell cycle, which could be suppressed by an inhibitor of miR‑20a. In conclusion, the data provide compelling evidence that miR‑20a functions as an onco‑miRNA, which is important in promoting cell proliferation in OS, and its oncogenic effect is mediated primarily through direct suppression of EGR2 expression.

Show MeSH
Related in: MedlinePlus