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Hairy/enhancer-of-split related with YRPW motif protein 1 promotes osteosarcoma metastasis via matrix metallopeptidase 9 expression.

Tsuru A, Setoguchi T, Matsunoshita Y, Nagao-Kitamoto H, Nagano S, Yokouchi M, Maeda S, Ishidou Y, Yamamoto T, Komiya S - Br. J. Cancer (2015)

Bottom Line: In addition, lung metastases were significantly inhibited by the knockdown of HEY1.We found that MMP9 was a downstream effector of HEY1 that promotes the invasion of osteosarcoma cells.Knockdown of HEY1 decreased the expression of MMP9.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8520, Japan.

ABSTRACT

Background: Activation of the Notch pathway has been reported in various types of cancers. However, the role of the hairy/enhancer-of-split related with YRPW motif protein 1 (HEY1) in osteosarcoma is unknown. We examined the function of HEY1 in osteosarcoma.

Methods: Expression of HEY1 was studied in human osteosarcoma. The effects of HEY1 in osteosarcoma were evaluated in vitro and in a xenograft model. Moreover, we examined the function of matrix metallopeptidase 9 (MMP9) as a downstream effector of HEY1.

Results: HEY1 was upregulated in human osteosarcoma. Knockdown of HEY1 inhibited the invasion of osteosarcoma cell lines. In contrast, the forced expression of HEY1 increased the invasion of mesenchymal stem cell. In addition, lung metastases were significantly inhibited by the knockdown of HEY1. We found that MMP9 was a downstream effector of HEY1 that promotes the invasion of osteosarcoma cells. Knockdown of HEY1 decreased the expression of MMP9. Addition of MMP9 rescued the invasion of osteosarcoma cells that had been rendered less invasive by knockdown of HEY1 expression.

Conclusions: Our findings suggested that HEY1 augmented the metastasis of osteosarcoma via upregulation of MMP9 expression. Therefore, inhibition of HEY1 may be a novel therapeutic strategy for preventing osteosarcoma metastasis.

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Related in: MedlinePlus

HEY1 promotes invasion of osteosarcoma cells.HEY1 siRNA-treated osteosarcoma cell lines or control siRNA-treated osteosarcoma cells were examined by using an invasion assay. Treatment with HEY1 siRNA decreased the expression of HEY1 mRNA. Column graphs show percent of invading cells after the invasion assay. Percent of invading cells in control siRNA-transfected cell lines were defined relative to an invasion of 100%. Representative photomicrographs show invaded cells (lower). Scale bar is 200 μm. *P< 0.05, Student's t-test.
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fig2: HEY1 promotes invasion of osteosarcoma cells.HEY1 siRNA-treated osteosarcoma cell lines or control siRNA-treated osteosarcoma cells were examined by using an invasion assay. Treatment with HEY1 siRNA decreased the expression of HEY1 mRNA. Column graphs show percent of invading cells after the invasion assay. Percent of invading cells in control siRNA-transfected cell lines were defined relative to an invasion of 100%. Representative photomicrographs show invaded cells (lower). Scale bar is 200 μm. *P< 0.05, Student's t-test.

Mentions: To examine the function of HEY1 and HEY2 in human osteosarcoma, we examined cell proliferation by using the WST-1 assay. Knockdown of HEY1 by siRNA (sc-37913: Santa Cruz Biotechnology) decreased the expression of HEY1 mRNA and protein (Figure 2 and Supplementary Figure 1A, B). Knockdown of HEY1 or HEY2 did not affect the proliferation of human osteosarcoma cells (Supplementary Figure 1B). The cell invasion assay showed that the knockdown of HEY1 prevented 143B osteosarcoma cell invasion (Figure 2A). To rule out the possibility of off-target effects, we examined an alternative HEY1-specific siRNA (LQ-008709-00-0002: Thermo Fisher Scientific) which also inhibited invasion (Supplementary Figure 2A). In addition, knockdown of HEY1 prevented HS-Os-1 and MG63 osteosarcoma cell invasion (Figure 2B and C). Knockdown of HEY2, however, did not affect the invasion of osteosarcoma cells (data not shown).


Hairy/enhancer-of-split related with YRPW motif protein 1 promotes osteosarcoma metastasis via matrix metallopeptidase 9 expression.

Tsuru A, Setoguchi T, Matsunoshita Y, Nagao-Kitamoto H, Nagano S, Yokouchi M, Maeda S, Ishidou Y, Yamamoto T, Komiya S - Br. J. Cancer (2015)

HEY1 promotes invasion of osteosarcoma cells.HEY1 siRNA-treated osteosarcoma cell lines or control siRNA-treated osteosarcoma cells were examined by using an invasion assay. Treatment with HEY1 siRNA decreased the expression of HEY1 mRNA. Column graphs show percent of invading cells after the invasion assay. Percent of invading cells in control siRNA-transfected cell lines were defined relative to an invasion of 100%. Representative photomicrographs show invaded cells (lower). Scale bar is 200 μm. *P< 0.05, Student's t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4385965&req=5

fig2: HEY1 promotes invasion of osteosarcoma cells.HEY1 siRNA-treated osteosarcoma cell lines or control siRNA-treated osteosarcoma cells were examined by using an invasion assay. Treatment with HEY1 siRNA decreased the expression of HEY1 mRNA. Column graphs show percent of invading cells after the invasion assay. Percent of invading cells in control siRNA-transfected cell lines were defined relative to an invasion of 100%. Representative photomicrographs show invaded cells (lower). Scale bar is 200 μm. *P< 0.05, Student's t-test.
Mentions: To examine the function of HEY1 and HEY2 in human osteosarcoma, we examined cell proliferation by using the WST-1 assay. Knockdown of HEY1 by siRNA (sc-37913: Santa Cruz Biotechnology) decreased the expression of HEY1 mRNA and protein (Figure 2 and Supplementary Figure 1A, B). Knockdown of HEY1 or HEY2 did not affect the proliferation of human osteosarcoma cells (Supplementary Figure 1B). The cell invasion assay showed that the knockdown of HEY1 prevented 143B osteosarcoma cell invasion (Figure 2A). To rule out the possibility of off-target effects, we examined an alternative HEY1-specific siRNA (LQ-008709-00-0002: Thermo Fisher Scientific) which also inhibited invasion (Supplementary Figure 2A). In addition, knockdown of HEY1 prevented HS-Os-1 and MG63 osteosarcoma cell invasion (Figure 2B and C). Knockdown of HEY2, however, did not affect the invasion of osteosarcoma cells (data not shown).

Bottom Line: In addition, lung metastases were significantly inhibited by the knockdown of HEY1.We found that MMP9 was a downstream effector of HEY1 that promotes the invasion of osteosarcoma cells.Knockdown of HEY1 decreased the expression of MMP9.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8520, Japan.

ABSTRACT

Background: Activation of the Notch pathway has been reported in various types of cancers. However, the role of the hairy/enhancer-of-split related with YRPW motif protein 1 (HEY1) in osteosarcoma is unknown. We examined the function of HEY1 in osteosarcoma.

Methods: Expression of HEY1 was studied in human osteosarcoma. The effects of HEY1 in osteosarcoma were evaluated in vitro and in a xenograft model. Moreover, we examined the function of matrix metallopeptidase 9 (MMP9) as a downstream effector of HEY1.

Results: HEY1 was upregulated in human osteosarcoma. Knockdown of HEY1 inhibited the invasion of osteosarcoma cell lines. In contrast, the forced expression of HEY1 increased the invasion of mesenchymal stem cell. In addition, lung metastases were significantly inhibited by the knockdown of HEY1. We found that MMP9 was a downstream effector of HEY1 that promotes the invasion of osteosarcoma cells. Knockdown of HEY1 decreased the expression of MMP9. Addition of MMP9 rescued the invasion of osteosarcoma cells that had been rendered less invasive by knockdown of HEY1 expression.

Conclusions: Our findings suggested that HEY1 augmented the metastasis of osteosarcoma via upregulation of MMP9 expression. Therefore, inhibition of HEY1 may be a novel therapeutic strategy for preventing osteosarcoma metastasis.

Show MeSH
Related in: MedlinePlus