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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

Bioluminescent imaging of tumor angiogenesis and alterations of tumor volume.(A) Bioluminescent imaging of tumor-bearing mice was obtained at 0 d, 3 d, 5 d, 7 d and 14 d in the same imaging conditions. (B) The dynamic measurements of bioluminescent intensity in tumors treated with saline, miR-497 mimic and miR-497 mimic-NC. Regions of interest (ROI) from displayed images revealed on the tumor sites and quantified as maximum photons per second per centimeter squared per steradian (P/s/cm2/sr). Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group. (C) Tumor volume curves of mice treated with saline, miR-497 mimic and miR-497 mimic-NC. Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group.
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f3: Bioluminescent imaging of tumor angiogenesis and alterations of tumor volume.(A) Bioluminescent imaging of tumor-bearing mice was obtained at 0 d, 3 d, 5 d, 7 d and 14 d in the same imaging conditions. (B) The dynamic measurements of bioluminescent intensity in tumors treated with saline, miR-497 mimic and miR-497 mimic-NC. Regions of interest (ROI) from displayed images revealed on the tumor sites and quantified as maximum photons per second per centimeter squared per steradian (P/s/cm2/sr). Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group. (C) Tumor volume curves of mice treated with saline, miR-497 mimic and miR-497 mimic-NC. Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group.

Mentions: For real-time visualization of the anti-angiogenesis effects of miR-497 via targeting VEGFR2, we used VEGFR2-luc transgenic mice to monitor tumor growth and angiogenesis in vivo by using BLI. After tumor cells injection for seven days, the VEGFR2-luc transgenic mice with tumor were randomized into three groups with the similar mean tumor volume: control group, miR-497 mimic group, miR-497 mimic-NC group. We defined the miR-497 mimic treatment day as 0 d, and as shown in Fig. 3A, there was no significant difference of tumor volume and bioluminescence signal intensity among three groups at 0 d (P > 0.05). With the increasing of tumor size, the bioluminescence signal was gradually enhanced in different treatment groups. Compared with the control, the bioluminescence signal intensity of miR-497 mimic treatment group was markedly lower on 5 d (P < 0.05) (Fig. 3A,B). Furthermore, the reduction of bioluminescence signal intensity was observed in miR-497 mimic group (~76%) compared to control on 14 d (P < 0.05). However, there was no statistical difference observed between control and miR-497 mimic-NC from 0 d to 14 d (P > 0.05) (Fig. 3A,B).


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)

Bioluminescent imaging of tumor angiogenesis and alterations of tumor volume.(A) Bioluminescent imaging of tumor-bearing mice was obtained at 0 d, 3 d, 5 d, 7 d and 14 d in the same imaging conditions. (B) The dynamic measurements of bioluminescent intensity in tumors treated with saline, miR-497 mimic and miR-497 mimic-NC. Regions of interest (ROI) from displayed images revealed on the tumor sites and quantified as maximum photons per second per centimeter squared per steradian (P/s/cm2/sr). Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group. (C) Tumor volume curves of mice treated with saline, miR-497 mimic and miR-497 mimic-NC. Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Bioluminescent imaging of tumor angiogenesis and alterations of tumor volume.(A) Bioluminescent imaging of tumor-bearing mice was obtained at 0 d, 3 d, 5 d, 7 d and 14 d in the same imaging conditions. (B) The dynamic measurements of bioluminescent intensity in tumors treated with saline, miR-497 mimic and miR-497 mimic-NC. Regions of interest (ROI) from displayed images revealed on the tumor sites and quantified as maximum photons per second per centimeter squared per steradian (P/s/cm2/sr). Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group. (C) Tumor volume curves of mice treated with saline, miR-497 mimic and miR-497 mimic-NC. Data were expressed as mean ± SEM, n = 5; **P < 0.01 vs. control group.
Mentions: For real-time visualization of the anti-angiogenesis effects of miR-497 via targeting VEGFR2, we used VEGFR2-luc transgenic mice to monitor tumor growth and angiogenesis in vivo by using BLI. After tumor cells injection for seven days, the VEGFR2-luc transgenic mice with tumor were randomized into three groups with the similar mean tumor volume: control group, miR-497 mimic group, miR-497 mimic-NC group. We defined the miR-497 mimic treatment day as 0 d, and as shown in Fig. 3A, there was no significant difference of tumor volume and bioluminescence signal intensity among three groups at 0 d (P > 0.05). With the increasing of tumor size, the bioluminescence signal was gradually enhanced in different treatment groups. Compared with the control, the bioluminescence signal intensity of miR-497 mimic treatment group was markedly lower on 5 d (P < 0.05) (Fig. 3A,B). Furthermore, the reduction of bioluminescence signal intensity was observed in miR-497 mimic group (~76%) compared to control on 14 d (P < 0.05). However, there was no statistical difference observed between control and miR-497 mimic-NC from 0 d to 14 d (P > 0.05) (Fig. 3A,B).

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