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Overexpression of miRNA-497 inhibits tumor angiogenesis by targeting VEGFR2.

Tu Y, Liu L, Zhao D, Liu Y, Ma X, Fan Y, Wan L, Huang T, Cheng Z, Shen B - Sci Rep (2015)

Bottom Line: The results demonstrated that overexpression of miR-497 showed inhibitory effects on VEGFR2 activation and downstream Raf/MEK/ERK signal pathways in vitro and in vivo.Moreover, overexpression of miR-497 effectively induced HUVECs apoptosis by targeting VEGFR2 and downstream PI3K/AKT signaling pathway.Furthermore, miR-497 exhibited anti-angiogenesis and anti-tumor effects in the VEGFR2-luc breast tumor model proven by BLI, WB and immunohistochemistry analysis.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Molecular Imaging, College of Heilongjiang Province, Harbin, Heilongjiang, China.

ABSTRACT
Recent studies reported miR-497 exhibited inhibitory effects in various cancers. However, whether miR-497 is involved in inhibiting angiogenesis, which is critical for tumor growth and metastasis, is still unknown. The purpose of this study was to investigate the potential role of miR-497 in tumor angiogenesis. In this work, cell proliferation and apoptosis analyses were conducted to explore the potential function of miR-497 in HUVECs by using MTT and TUNEL assays. Western blotting (WB) was employed to validate the downstream targets of miR-497. Furthermore, in order to disclose the role of miR-497 on angiogenesis, VEGFR2-luc transgenic mice were treated with miR-497 mimic and applied to monitor tumor angiogenesis and growth by in vivo bioluminescent imaging (BLI). The results demonstrated that overexpression of miR-497 showed inhibitory effects on VEGFR2 activation and downstream Raf/MEK/ERK signal pathways in vitro and in vivo. Moreover, overexpression of miR-497 effectively induced HUVECs apoptosis by targeting VEGFR2 and downstream PI3K/AKT signaling pathway. Furthermore, miR-497 exhibited anti-angiogenesis and anti-tumor effects in the VEGFR2-luc breast tumor model proven by BLI, WB and immunohistochemistry analysis. In summary, miR-497 inhibits tumor angiogenesis and growth via targeting VEGFR2, indicating miR-497 can be explored as a potential drug candidate for cancer therapy.

No MeSH data available.


Related in: MedlinePlus

Overexpression of miR-497 influenced the proliferation of cultured HUVECs.(A) Cell proliferation was evaluated by the MTT assay; (B–F) Protein levels of Raf, Total-MEK, p-MEK, Total-ERK and p-ERK were determined by western blotting assay. Data were expressed as mean ± SEM, n = 3; **P < 0.01 or ***P < 0.001 vs. control group.
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f2: Overexpression of miR-497 influenced the proliferation of cultured HUVECs.(A) Cell proliferation was evaluated by the MTT assay; (B–F) Protein levels of Raf, Total-MEK, p-MEK, Total-ERK and p-ERK were determined by western blotting assay. Data were expressed as mean ± SEM, n = 3; **P < 0.01 or ***P < 0.001 vs. control group.

Mentions: Angiogenesis is a complex process orchestrated by both growth factors and cell adhesion, and it is initiated by focal degradation of the vascular basement membrane with subsequent migration and proliferation of endothelial cells. The Raf/MEK/ERK pathway is required for endothelial cell function during angiogenesis. To explore the potential impact of miR-497 on the proliferation of HUVECs, cellular viability and proliferation were measured following the protocol of MTT and western blotting assay. The results showed that HUVECs transfected with miR-497 mimic, but not miR-497 inhibitor, proliferated at a lower rate as compared with control group (P < 0.05) (Fig. 2A), indicating that up-regulation of miR-497 could suppress the growth of HUVECs. However, miR-497 mimic-NC could not play the same role (P > 0.05) (Fig. 2A).


Overexpression of miRNA-497 inhibits tumor angiogenesis by targeting VEGFR2.

Tu Y, Liu L, Zhao D, Liu Y, Ma X, Fan Y, Wan L, Huang T, Cheng Z, Shen B - Sci Rep (2015)

Overexpression of miR-497 influenced the proliferation of cultured HUVECs.(A) Cell proliferation was evaluated by the MTT assay; (B–F) Protein levels of Raf, Total-MEK, p-MEK, Total-ERK and p-ERK were determined by western blotting assay. Data were expressed as mean ± SEM, n = 3; **P < 0.01 or ***P < 0.001 vs. control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561885&req=5

f2: Overexpression of miR-497 influenced the proliferation of cultured HUVECs.(A) Cell proliferation was evaluated by the MTT assay; (B–F) Protein levels of Raf, Total-MEK, p-MEK, Total-ERK and p-ERK were determined by western blotting assay. Data were expressed as mean ± SEM, n = 3; **P < 0.01 or ***P < 0.001 vs. control group.
Mentions: Angiogenesis is a complex process orchestrated by both growth factors and cell adhesion, and it is initiated by focal degradation of the vascular basement membrane with subsequent migration and proliferation of endothelial cells. The Raf/MEK/ERK pathway is required for endothelial cell function during angiogenesis. To explore the potential impact of miR-497 on the proliferation of HUVECs, cellular viability and proliferation were measured following the protocol of MTT and western blotting assay. The results showed that HUVECs transfected with miR-497 mimic, but not miR-497 inhibitor, proliferated at a lower rate as compared with control group (P < 0.05) (Fig. 2A), indicating that up-regulation of miR-497 could suppress the growth of HUVECs. However, miR-497 mimic-NC could not play the same role (P > 0.05) (Fig. 2A).

Bottom Line: The results demonstrated that overexpression of miR-497 showed inhibitory effects on VEGFR2 activation and downstream Raf/MEK/ERK signal pathways in vitro and in vivo.Moreover, overexpression of miR-497 effectively induced HUVECs apoptosis by targeting VEGFR2 and downstream PI3K/AKT signaling pathway.Furthermore, miR-497 exhibited anti-angiogenesis and anti-tumor effects in the VEGFR2-luc breast tumor model proven by BLI, WB and immunohistochemistry analysis.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Molecular Imaging, College of Heilongjiang Province, Harbin, Heilongjiang, China.

ABSTRACT
Recent studies reported miR-497 exhibited inhibitory effects in various cancers. However, whether miR-497 is involved in inhibiting angiogenesis, which is critical for tumor growth and metastasis, is still unknown. The purpose of this study was to investigate the potential role of miR-497 in tumor angiogenesis. In this work, cell proliferation and apoptosis analyses were conducted to explore the potential function of miR-497 in HUVECs by using MTT and TUNEL assays. Western blotting (WB) was employed to validate the downstream targets of miR-497. Furthermore, in order to disclose the role of miR-497 on angiogenesis, VEGFR2-luc transgenic mice were treated with miR-497 mimic and applied to monitor tumor angiogenesis and growth by in vivo bioluminescent imaging (BLI). The results demonstrated that overexpression of miR-497 showed inhibitory effects on VEGFR2 activation and downstream Raf/MEK/ERK signal pathways in vitro and in vivo. Moreover, overexpression of miR-497 effectively induced HUVECs apoptosis by targeting VEGFR2 and downstream PI3K/AKT signaling pathway. Furthermore, miR-497 exhibited anti-angiogenesis and anti-tumor effects in the VEGFR2-luc breast tumor model proven by BLI, WB and immunohistochemistry analysis. In summary, miR-497 inhibits tumor angiogenesis and growth via targeting VEGFR2, indicating miR-497 can be explored as a potential drug candidate for cancer therapy.

No MeSH data available.


Related in: MedlinePlus