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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

Representative CD31, CD61 and VEGFR2 immunostaining images were carried out under different experimental conditions as indicated (Original magnification, ×200).(A) CD31 was stained with red color and CD61 was stained with green color; (B) MVD of tumors in each group was detected by the vessel counts. MVD: micro-vessel density, HPF: high-power field. Data were expressed as mean ± SEM, n = 10; **P < 0.01 vs. control group; (C) VEGFR2 was stained with green color, and cells nuclei were blue.
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f5: Representative CD31, CD61 and VEGFR2 immunostaining images were carried out under different experimental conditions as indicated (Original magnification, ×200).(A) CD31 was stained with red color and CD61 was stained with green color; (B) MVD of tumors in each group was detected by the vessel counts. MVD: micro-vessel density, HPF: high-power field. Data were expressed as mean ± SEM, n = 10; **P < 0.01 vs. control group; (C) VEGFR2 was stained with green color, and cells nuclei were blue.

Mentions: In this study, immunofluorescent staining with CD31 and CD61 was applied to testify MVD in the tumor sections. The results demonstrated that the mean MVD of control, miR-497 mimic and miR-497 mimic-NC groups was 76 ± 1.6, 35 ± 1.2 and 82 ± 3.5, respectively. The tumor sample from miR-497 mimic group had less capillary blood vessels than the control group (35 ± 1.2 vs. 76 ± 1.6, P < 0.05) (Fig. 5A,B), while compared with the control, MVD of tumor section in miR-497 mimic-NC group showed no statistical difference (82 ± 3.5 vs. 76 ± 1.6, P > 0.05) (Fig. 5A,B). More importantly, the negative correlation between miR-497 expression and MVD of tumor tissues was found in this study, which suggested that the miR-497 played a vital role in suppressing tumor angiogenesis.


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)

Representative CD31, CD61 and VEGFR2 immunostaining images were carried out under different experimental conditions as indicated (Original magnification, ×200).(A) CD31 was stained with red color and CD61 was stained with green color; (B) MVD of tumors in each group was detected by the vessel counts. MVD: micro-vessel density, HPF: high-power field. Data were expressed as mean ± SEM, n = 10; **P < 0.01 vs. control group; (C) VEGFR2 was stained with green color, and cells nuclei were blue.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Representative CD31, CD61 and VEGFR2 immunostaining images were carried out under different experimental conditions as indicated (Original magnification, ×200).(A) CD31 was stained with red color and CD61 was stained with green color; (B) MVD of tumors in each group was detected by the vessel counts. MVD: micro-vessel density, HPF: high-power field. Data were expressed as mean ± SEM, n = 10; **P < 0.01 vs. control group; (C) VEGFR2 was stained with green color, and cells nuclei were blue.
Mentions: In this study, immunofluorescent staining with CD31 and CD61 was applied to testify MVD in the tumor sections. The results demonstrated that the mean MVD of control, miR-497 mimic and miR-497 mimic-NC groups was 76 ± 1.6, 35 ± 1.2 and 82 ± 3.5, respectively. The tumor sample from miR-497 mimic group had less capillary blood vessels than the control group (35 ± 1.2 vs. 76 ± 1.6, P < 0.05) (Fig. 5A,B), while compared with the control, MVD of tumor section in miR-497 mimic-NC group showed no statistical difference (82 ± 3.5 vs. 76 ± 1.6, P > 0.05) (Fig. 5A,B). More importantly, the negative correlation between miR-497 expression and MVD of tumor tissues was found in this study, which suggested that the miR-497 played a vital role in suppressing tumor angiogenesis.

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