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NOTCH1 signaling contributes to cell growth, anti-apoptosis and metastasis in salivary adenoid cystic carcinoma.

Su BH, Qu J, Song M, Huang XY, Hu XM, Xie J, Zhao Y, Ding LC, She L, Chen J, Lin LS, Lin X, Zheng DL, Lu YG - Oncotarget (2014)

Bottom Line: We silenced the expression of NOTCH1 and overexpressed activated NOTCH1 to elucidate the effects of NOTCH1 on proliferation, migration and invasion.NOTCH1 target genes were validated by real-time PCR.Overexpression of NOTCH1 in SACC cells promoted cell growth, migration and invasion, and knockdown of NOTCH1 inhibited cell proliferation in vitro and tumorigenicity in vivo by inducing cell apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Preventive Dentistry, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China. These authors contributed equally to this work.

ABSTRACT

Background: Numerous studies have reported both the tumor-suppressive and oncogenic roles of the Notch pathway, indicating that Notch activity regulates tumor biology in a complex, context-dependent manner. The aim of the present study was to identify the role of NOTCH1 in the cell growth and metastasis of SACC.

Methods: We analyzed the expression of NOTCH1 in clinical SACC samples using immunohistochemical staining. We silenced the expression of NOTCH1 and overexpressed activated NOTCH1 to elucidate the effects of NOTCH1 on proliferation, migration and invasion. NOTCH1 target genes were validated by real-time PCR.

Results: Our results showed that NOTCH1 was upregulated in SACC tissues when compared with normal tissues, and this upregulation was further enhanced in SACC tissues with metastasis and recurrence when compared with SACC tissues without metastasis. Overexpression of NOTCH1 in SACC cells promoted cell growth, migration and invasion, and knockdown of NOTCH1 inhibited cell proliferation in vitro and tumorigenicity in vivo by inducing cell apoptosis.

Conclusions: The results of this study suggest that NOTCH1 plays a key role in the cell growth, anti-apoptosis, and metastasis of SACC. NOTCH1 inhibitors might therefore have potential therapeutic applications in treating SACC patients by inhibiting cancer cell growth and metastasis.

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Related in: MedlinePlus

Knockdown of NOTCH1 inhibits the migration and invasion abilities of SACC-83 cellsA, A photomicrograph of scratch wounds made in the siRNAs transfected SACC-83 cell layer shows inhibited cellular motility in the NOTCH1 silenced cells compared with the control. B, Representative images of the transwell assay without (upper panel) or with (lower panel) coated Matrigel after siRNA transfection. C, The number of cells that migrated through uncoated filters (i.e., no Matrigel), which represents the migratory ability of SACC-83 cells. D, The number of cells that were able to pass through filters precoated with Matrigel, which represents the invasive ability of SACC-83 cells. The counts of the cells are presented as the mean values per field from at least five randomly selected low-powered fields (x200) from three independent experiments (error bars, means ± SD). P<0.01 when compared with the control (NC).
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Figure 4: Knockdown of NOTCH1 inhibits the migration and invasion abilities of SACC-83 cellsA, A photomicrograph of scratch wounds made in the siRNAs transfected SACC-83 cell layer shows inhibited cellular motility in the NOTCH1 silenced cells compared with the control. B, Representative images of the transwell assay without (upper panel) or with (lower panel) coated Matrigel after siRNA transfection. C, The number of cells that migrated through uncoated filters (i.e., no Matrigel), which represents the migratory ability of SACC-83 cells. D, The number of cells that were able to pass through filters precoated with Matrigel, which represents the invasive ability of SACC-83 cells. The counts of the cells are presented as the mean values per field from at least five randomly selected low-powered fields (x200) from three independent experiments (error bars, means ± SD). P<0.01 when compared with the control (NC).

Mentions: Next, we asked whether NOTCH1 plays a role in the migration and invasion of SACC. As shown in Fig. 4, the knockdown of NOTCH1 in SACC-83 cells significantly inhibited cell migration (Fig. 4A, B and C, P<0.01, n=3) and invasion (Fig. 4B and C, P<0.01, n=3). In contrast, the overexpression of NICD1 in SACC-83 cells promoted cell motility, as indicated by the wound healing assay (Fig. 5A) and transwell assay (Fig. 5B and C, P<0.05, n=3), as well as cell invasiveness (Fig. 5B and C, P<0.05, n=3). These results confirm that NOTCH1 is an oncogene in SACC that may contribute to the migration and invasion of adenoid cystic carcinoma cells.


NOTCH1 signaling contributes to cell growth, anti-apoptosis and metastasis in salivary adenoid cystic carcinoma.

Su BH, Qu J, Song M, Huang XY, Hu XM, Xie J, Zhao Y, Ding LC, She L, Chen J, Lin LS, Lin X, Zheng DL, Lu YG - Oncotarget (2014)

Knockdown of NOTCH1 inhibits the migration and invasion abilities of SACC-83 cellsA, A photomicrograph of scratch wounds made in the siRNAs transfected SACC-83 cell layer shows inhibited cellular motility in the NOTCH1 silenced cells compared with the control. B, Representative images of the transwell assay without (upper panel) or with (lower panel) coated Matrigel after siRNA transfection. C, The number of cells that migrated through uncoated filters (i.e., no Matrigel), which represents the migratory ability of SACC-83 cells. D, The number of cells that were able to pass through filters precoated with Matrigel, which represents the invasive ability of SACC-83 cells. The counts of the cells are presented as the mean values per field from at least five randomly selected low-powered fields (x200) from three independent experiments (error bars, means ± SD). P<0.01 when compared with the control (NC).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196170&req=5

Figure 4: Knockdown of NOTCH1 inhibits the migration and invasion abilities of SACC-83 cellsA, A photomicrograph of scratch wounds made in the siRNAs transfected SACC-83 cell layer shows inhibited cellular motility in the NOTCH1 silenced cells compared with the control. B, Representative images of the transwell assay without (upper panel) or with (lower panel) coated Matrigel after siRNA transfection. C, The number of cells that migrated through uncoated filters (i.e., no Matrigel), which represents the migratory ability of SACC-83 cells. D, The number of cells that were able to pass through filters precoated with Matrigel, which represents the invasive ability of SACC-83 cells. The counts of the cells are presented as the mean values per field from at least five randomly selected low-powered fields (x200) from three independent experiments (error bars, means ± SD). P<0.01 when compared with the control (NC).
Mentions: Next, we asked whether NOTCH1 plays a role in the migration and invasion of SACC. As shown in Fig. 4, the knockdown of NOTCH1 in SACC-83 cells significantly inhibited cell migration (Fig. 4A, B and C, P<0.01, n=3) and invasion (Fig. 4B and C, P<0.01, n=3). In contrast, the overexpression of NICD1 in SACC-83 cells promoted cell motility, as indicated by the wound healing assay (Fig. 5A) and transwell assay (Fig. 5B and C, P<0.05, n=3), as well as cell invasiveness (Fig. 5B and C, P<0.05, n=3). These results confirm that NOTCH1 is an oncogene in SACC that may contribute to the migration and invasion of adenoid cystic carcinoma cells.

Bottom Line: We silenced the expression of NOTCH1 and overexpressed activated NOTCH1 to elucidate the effects of NOTCH1 on proliferation, migration and invasion.NOTCH1 target genes were validated by real-time PCR.Overexpression of NOTCH1 in SACC cells promoted cell growth, migration and invasion, and knockdown of NOTCH1 inhibited cell proliferation in vitro and tumorigenicity in vivo by inducing cell apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Preventive Dentistry, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China. These authors contributed equally to this work.

ABSTRACT

Background: Numerous studies have reported both the tumor-suppressive and oncogenic roles of the Notch pathway, indicating that Notch activity regulates tumor biology in a complex, context-dependent manner. The aim of the present study was to identify the role of NOTCH1 in the cell growth and metastasis of SACC.

Methods: We analyzed the expression of NOTCH1 in clinical SACC samples using immunohistochemical staining. We silenced the expression of NOTCH1 and overexpressed activated NOTCH1 to elucidate the effects of NOTCH1 on proliferation, migration and invasion. NOTCH1 target genes were validated by real-time PCR.

Results: Our results showed that NOTCH1 was upregulated in SACC tissues when compared with normal tissues, and this upregulation was further enhanced in SACC tissues with metastasis and recurrence when compared with SACC tissues without metastasis. Overexpression of NOTCH1 in SACC cells promoted cell growth, migration and invasion, and knockdown of NOTCH1 inhibited cell proliferation in vitro and tumorigenicity in vivo by inducing cell apoptosis.

Conclusions: The results of this study suggest that NOTCH1 plays a key role in the cell growth, anti-apoptosis, and metastasis of SACC. NOTCH1 inhibitors might therefore have potential therapeutic applications in treating SACC patients by inhibiting cancer cell growth and metastasis.

Show MeSH
Related in: MedlinePlus