Limits...
MicroRNA-138 functions as a tumor suppressor in osteosarcoma by targeting differentiated embryonic chondrocyte gene 2.

Jiang B, Mu W, Wang J, Lu J, Jiang S, Li L, Xu H, Tian H - J. Exp. Clin. Cancer Res. (2016)

Bottom Line: MicroRNA-138 (miR-138) has been proven to be a tumor suppressor gene in various types of tumors.Moreover, miR-138 expression was significantly lower in metastatic osteosarcoma tissues than that in non-metastatic tissues.DEC2 was verified as a direct target of miR-138, and DEC2 could reverse the inhibitory effect of miR-138 on osteosarcoma progression.

View Article: PubMed Central - PubMed

Affiliation: Department of Traumatic Orthopaedics, The People 's Hospital of Dongying City of Shandong Province, No 317 Nanyi Road, Dongying, 257091, Shandong, China. baoen_jiang@163.com.

ABSTRACT

Background: MicroRNA-138 (miR-138) has been proven to be a tumor suppressor gene in various types of tumors. However, the expression and the role of miR-138 in human osteosarcoma are still poorly understood. We investigated the function and the underlying mechanism of miR-138 in osteosarcoma.

Methods: The expression of miR-138 in human osteosarcoma tissues and cell lines was detected by real-time PCR analysis. The gain-of-function and loss-of-function experiments were performed on osteosarcoma cell lines to investigate the effects of miR-138 on osteosarcoma progression, and to determine whether differentiated embryonic chondrocyte gene 2 (DEC2) mediates these effects. Cell proliferation, apoptosis and invasion were assessed by MTT, flow cytometry and transwell-matrigel assays. Dual-luciferase reporter assay was used to identify whether DEC2 is a direct target of miR-138.

Results: MiR-138 was significantly downregulated in human osteosarcoma tissues and cell lines. Moreover, miR-138 expression was significantly lower in metastatic osteosarcoma tissues than that in non-metastatic tissues. The in vitro gain-of-function and loss-of-function experiments demonstrated that miR-138 inhibited cell proliferation and invasion, and promoted cell apoptosis of human osteosarcoma cells. DEC2 was verified as a direct target of miR-138, and DEC2 could reverse the inhibitory effect of miR-138 on osteosarcoma progression.

Conclusions: These findings suggested that miR-138 acts as a tumor suppressor in osteosarcoma.miR-138 inhibited cell proliferation and invasion, as well as promoted cell apoptosis of human osteosarcoma cells, at least partially, by inhibiting the expression of DEC2. MiR-138/DEC2 may be a novel therapeutic target in osteosarcoma.

No MeSH data available.


Related in: MedlinePlus

DEC2 was a direct target of miR-138. a The DEC2 3′UTR region containing the wild type or mutant binding site for miR-138. b The relative luciferase activity of DEC2 wild type or mutant 3'UTR in HEK293 cells following transfection with the miR-138 mimic. c Fold change of DEC2 mRNA expression and relative DEC2 protein expression in U2OS cells following transfection with the miR-138 mimic. Lane 1, miR-Ctrl; lane 2, miR-138 mimic. d Fold change of DEC2 mRNA expression and relative DEC2 protein expression in MG-63 cells following transfection with the miR-138 inhibitor. Lane 1, miR-Ctrl; lane 2, miR-138 inhibitor. *P < 0.05 and #P < 0.01 compared with the miR-Ctrl
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4837633&req=5

Fig4: DEC2 was a direct target of miR-138. a The DEC2 3′UTR region containing the wild type or mutant binding site for miR-138. b The relative luciferase activity of DEC2 wild type or mutant 3'UTR in HEK293 cells following transfection with the miR-138 mimic. c Fold change of DEC2 mRNA expression and relative DEC2 protein expression in U2OS cells following transfection with the miR-138 mimic. Lane 1, miR-Ctrl; lane 2, miR-138 mimic. d Fold change of DEC2 mRNA expression and relative DEC2 protein expression in MG-63 cells following transfection with the miR-138 inhibitor. Lane 1, miR-Ctrl; lane 2, miR-138 inhibitor. *P < 0.05 and #P < 0.01 compared with the miR-Ctrl

Mentions: DEC2 is predicted to be a potential target of miR-138 by miRanda (http://www.microrna.org/). The predicted interaction site of 7 bp of miR-138 and DEC2 3′UTR was shown in Fig. 4a. To identify whether DEC2 is a direct target of miR-138, wild type and mutant DEC2 3′UTR containing the putative target site of miR-138 were cloned into reporter plasmids respectively, and were transfected into the HEK293 cells along with the miR-138 mimic, or the control miRNA. As validated by luciferase reporter assay, the luciferase activity of wild type DEC2-3′UTR was significantly suppressed in the cells transfected with the miR-138 mimic compared with the cells transfected with the miR-Ctrl (P < 0.01); however, miR-138 mimic did not affect the luciferase activity of mutant DEC2-3′UTR (Fig.4b).Fig 4


MicroRNA-138 functions as a tumor suppressor in osteosarcoma by targeting differentiated embryonic chondrocyte gene 2.

Jiang B, Mu W, Wang J, Lu J, Jiang S, Li L, Xu H, Tian H - J. Exp. Clin. Cancer Res. (2016)

DEC2 was a direct target of miR-138. a The DEC2 3′UTR region containing the wild type or mutant binding site for miR-138. b The relative luciferase activity of DEC2 wild type or mutant 3'UTR in HEK293 cells following transfection with the miR-138 mimic. c Fold change of DEC2 mRNA expression and relative DEC2 protein expression in U2OS cells following transfection with the miR-138 mimic. Lane 1, miR-Ctrl; lane 2, miR-138 mimic. d Fold change of DEC2 mRNA expression and relative DEC2 protein expression in MG-63 cells following transfection with the miR-138 inhibitor. Lane 1, miR-Ctrl; lane 2, miR-138 inhibitor. *P < 0.05 and #P < 0.01 compared with the miR-Ctrl
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4837633&req=5

Fig4: DEC2 was a direct target of miR-138. a The DEC2 3′UTR region containing the wild type or mutant binding site for miR-138. b The relative luciferase activity of DEC2 wild type or mutant 3'UTR in HEK293 cells following transfection with the miR-138 mimic. c Fold change of DEC2 mRNA expression and relative DEC2 protein expression in U2OS cells following transfection with the miR-138 mimic. Lane 1, miR-Ctrl; lane 2, miR-138 mimic. d Fold change of DEC2 mRNA expression and relative DEC2 protein expression in MG-63 cells following transfection with the miR-138 inhibitor. Lane 1, miR-Ctrl; lane 2, miR-138 inhibitor. *P < 0.05 and #P < 0.01 compared with the miR-Ctrl
Mentions: DEC2 is predicted to be a potential target of miR-138 by miRanda (http://www.microrna.org/). The predicted interaction site of 7 bp of miR-138 and DEC2 3′UTR was shown in Fig. 4a. To identify whether DEC2 is a direct target of miR-138, wild type and mutant DEC2 3′UTR containing the putative target site of miR-138 were cloned into reporter plasmids respectively, and were transfected into the HEK293 cells along with the miR-138 mimic, or the control miRNA. As validated by luciferase reporter assay, the luciferase activity of wild type DEC2-3′UTR was significantly suppressed in the cells transfected with the miR-138 mimic compared with the cells transfected with the miR-Ctrl (P < 0.01); however, miR-138 mimic did not affect the luciferase activity of mutant DEC2-3′UTR (Fig.4b).Fig 4

Bottom Line: MicroRNA-138 (miR-138) has been proven to be a tumor suppressor gene in various types of tumors.Moreover, miR-138 expression was significantly lower in metastatic osteosarcoma tissues than that in non-metastatic tissues.DEC2 was verified as a direct target of miR-138, and DEC2 could reverse the inhibitory effect of miR-138 on osteosarcoma progression.

View Article: PubMed Central - PubMed

Affiliation: Department of Traumatic Orthopaedics, The People 's Hospital of Dongying City of Shandong Province, No 317 Nanyi Road, Dongying, 257091, Shandong, China. baoen_jiang@163.com.

ABSTRACT

Background: MicroRNA-138 (miR-138) has been proven to be a tumor suppressor gene in various types of tumors. However, the expression and the role of miR-138 in human osteosarcoma are still poorly understood. We investigated the function and the underlying mechanism of miR-138 in osteosarcoma.

Methods: The expression of miR-138 in human osteosarcoma tissues and cell lines was detected by real-time PCR analysis. The gain-of-function and loss-of-function experiments were performed on osteosarcoma cell lines to investigate the effects of miR-138 on osteosarcoma progression, and to determine whether differentiated embryonic chondrocyte gene 2 (DEC2) mediates these effects. Cell proliferation, apoptosis and invasion were assessed by MTT, flow cytometry and transwell-matrigel assays. Dual-luciferase reporter assay was used to identify whether DEC2 is a direct target of miR-138.

Results: MiR-138 was significantly downregulated in human osteosarcoma tissues and cell lines. Moreover, miR-138 expression was significantly lower in metastatic osteosarcoma tissues than that in non-metastatic tissues. The in vitro gain-of-function and loss-of-function experiments demonstrated that miR-138 inhibited cell proliferation and invasion, and promoted cell apoptosis of human osteosarcoma cells. DEC2 was verified as a direct target of miR-138, and DEC2 could reverse the inhibitory effect of miR-138 on osteosarcoma progression.

Conclusions: These findings suggested that miR-138 acts as a tumor suppressor in osteosarcoma.miR-138 inhibited cell proliferation and invasion, as well as promoted cell apoptosis of human osteosarcoma cells, at least partially, by inhibiting the expression of DEC2. MiR-138/DEC2 may be a novel therapeutic target in osteosarcoma.

No MeSH data available.


Related in: MedlinePlus