Limits...
ADAM17 mediates OSCC development in an orthotopic murine model.

Simabuco FM, Kawahara R, Yokoo S, Granato DC, Miguel L, Agostini M, Aragão AZ, Domingues RR, Flores IL, Macedo CC, Della Coletta R, Graner E, Paes Leme AF - Mol. Cancer (2014)

Bottom Line: In this study the effect of overexpressing ADAM17 in cell migration, viability, adhesion and proliferation was comprehensively appraised in vitro.These findings contribute to understand the role of ADAM17 in oral cancer development and as a potential therapeutic target in oral cancer.In addition, our study also provides the basis for the development of novel and refined OSCC-targeting approaches.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratório de Espectrometria de Massas, Laboratório Nacional de Biociências, LNBio, CNPEM, Campinas 13083-970, Brazil. adriana.paesleme@lnbio.cnpem.br.

ABSTRACT

Background: ADAM17 is one of the main sheddases of the cells and it is responsible for the cleavage and the release of ectodomains of important signaling molecules, such as EGFR ligands. Despite the known crosstalk between ADAM17 and EGFR, which has been considered a promising targeted therapy in oral squamous cell carcinoma (OSCC), the role of ADAM17 in OSCC development is not clear.

Method: In this study the effect of overexpressing ADAM17 in cell migration, viability, adhesion and proliferation was comprehensively appraised in vitro. In addition, the tumor size, tumor proliferative activity, tumor collagenase activity and MS-based proteomics of tumor tissues have been evaluated by injecting tumorigenic squamous carcinoma cells (SCC-9) overexpressing ADAM17 in immunodeficient mice.

Results: The proteomic analysis has effectively identified a total of 2,194 proteins in control and tumor tissues. Among these, 110 proteins have been down-regulated and 90 have been up-regulated in tumor tissues. Biological network analysis has uncovered that overexpression of ADAM17 regulates Erk pathway in OSCC and further indicates proteins regulated by the overexpression of ADAM17 in the respective pathway. These results are also supported by the evidences of higher viability, migration, adhesion and proliferation in SCC-9 or A431 cells in vitro along with the increase of tumor size and proliferative activity and higher tissue collagenase activity as an outcome of ADAM17 overexpression.

Conclusion: These findings contribute to understand the role of ADAM17 in oral cancer development and as a potential therapeutic target in oral cancer. In addition, our study also provides the basis for the development of novel and refined OSCC-targeting approaches.

Show MeSH

Related in: MedlinePlus

SCC-9 cells overexpressing ADAM17-HA increased tumor size and proliferation. (A) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce increased size of tumors in Nude mice. Cells grown in tissue culture plates were trypsinized, resuspended in PBS and injected in the tongue of Nude mice. After 20 days, mice were sacrificed, tumors excised and measured (n = 3, Student’s t-test, p = 0.0467). (B) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce higher proliferative activity by immunohistochemical expression of Ki-67. Positive cells were calculated by counting labeled nuclei (positive-cells) of six high power fields (magnification of 400×) from each case with the aid of the Image J software and expressing the data as percentage (n = 6, Student’s t-test, p < 0.0001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC3928084&req=5

Figure 3: SCC-9 cells overexpressing ADAM17-HA increased tumor size and proliferation. (A) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce increased size of tumors in Nude mice. Cells grown in tissue culture plates were trypsinized, resuspended in PBS and injected in the tongue of Nude mice. After 20 days, mice were sacrificed, tumors excised and measured (n = 3, Student’s t-test, p = 0.0467). (B) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce higher proliferative activity by immunohistochemical expression of Ki-67. Positive cells were calculated by counting labeled nuclei (positive-cells) of six high power fields (magnification of 400×) from each case with the aid of the Image J software and expressing the data as percentage (n = 6, Student’s t-test, p < 0.0001).

Mentions: An orthotopic murine tumor formation model using SCC-9 cells overexpressing ADAM17 or GFP was performed. After 20 days, tumors were excised and had their size measured. As seen in Figure 3A, tumors induced with SCC-9 cells overexpressing ADAM17 had increased size compared to SCC-9 GFP cells (n = 3, Student’s t-test, p = 0.0467). SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce higher proliferative activity by immunohistochemical expression of Ki-67 compared to SCC-9 GFP cells (n = 6, Student’s t-test, p < 0.0001) (Figure 3B).


ADAM17 mediates OSCC development in an orthotopic murine model.

Simabuco FM, Kawahara R, Yokoo S, Granato DC, Miguel L, Agostini M, Aragão AZ, Domingues RR, Flores IL, Macedo CC, Della Coletta R, Graner E, Paes Leme AF - Mol. Cancer (2014)

SCC-9 cells overexpressing ADAM17-HA increased tumor size and proliferation. (A) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce increased size of tumors in Nude mice. Cells grown in tissue culture plates were trypsinized, resuspended in PBS and injected in the tongue of Nude mice. After 20 days, mice were sacrificed, tumors excised and measured (n = 3, Student’s t-test, p = 0.0467). (B) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce higher proliferative activity by immunohistochemical expression of Ki-67. Positive cells were calculated by counting labeled nuclei (positive-cells) of six high power fields (magnification of 400×) from each case with the aid of the Image J software and expressing the data as percentage (n = 6, Student’s t-test, p < 0.0001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3928084&req=5

Figure 3: SCC-9 cells overexpressing ADAM17-HA increased tumor size and proliferation. (A) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce increased size of tumors in Nude mice. Cells grown in tissue culture plates were trypsinized, resuspended in PBS and injected in the tongue of Nude mice. After 20 days, mice were sacrificed, tumors excised and measured (n = 3, Student’s t-test, p = 0.0467). (B) SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce higher proliferative activity by immunohistochemical expression of Ki-67. Positive cells were calculated by counting labeled nuclei (positive-cells) of six high power fields (magnification of 400×) from each case with the aid of the Image J software and expressing the data as percentage (n = 6, Student’s t-test, p < 0.0001).
Mentions: An orthotopic murine tumor formation model using SCC-9 cells overexpressing ADAM17 or GFP was performed. After 20 days, tumors were excised and had their size measured. As seen in Figure 3A, tumors induced with SCC-9 cells overexpressing ADAM17 had increased size compared to SCC-9 GFP cells (n = 3, Student’s t-test, p = 0.0467). SCC-9 cells overexpressing ADAM17-HA (AD17-HA) induce higher proliferative activity by immunohistochemical expression of Ki-67 compared to SCC-9 GFP cells (n = 6, Student’s t-test, p < 0.0001) (Figure 3B).

Bottom Line: In this study the effect of overexpressing ADAM17 in cell migration, viability, adhesion and proliferation was comprehensively appraised in vitro.These findings contribute to understand the role of ADAM17 in oral cancer development and as a potential therapeutic target in oral cancer.In addition, our study also provides the basis for the development of novel and refined OSCC-targeting approaches.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratório de Espectrometria de Massas, Laboratório Nacional de Biociências, LNBio, CNPEM, Campinas 13083-970, Brazil. adriana.paesleme@lnbio.cnpem.br.

ABSTRACT

Background: ADAM17 is one of the main sheddases of the cells and it is responsible for the cleavage and the release of ectodomains of important signaling molecules, such as EGFR ligands. Despite the known crosstalk between ADAM17 and EGFR, which has been considered a promising targeted therapy in oral squamous cell carcinoma (OSCC), the role of ADAM17 in OSCC development is not clear.

Method: In this study the effect of overexpressing ADAM17 in cell migration, viability, adhesion and proliferation was comprehensively appraised in vitro. In addition, the tumor size, tumor proliferative activity, tumor collagenase activity and MS-based proteomics of tumor tissues have been evaluated by injecting tumorigenic squamous carcinoma cells (SCC-9) overexpressing ADAM17 in immunodeficient mice.

Results: The proteomic analysis has effectively identified a total of 2,194 proteins in control and tumor tissues. Among these, 110 proteins have been down-regulated and 90 have been up-regulated in tumor tissues. Biological network analysis has uncovered that overexpression of ADAM17 regulates Erk pathway in OSCC and further indicates proteins regulated by the overexpression of ADAM17 in the respective pathway. These results are also supported by the evidences of higher viability, migration, adhesion and proliferation in SCC-9 or A431 cells in vitro along with the increase of tumor size and proliferative activity and higher tissue collagenase activity as an outcome of ADAM17 overexpression.

Conclusion: These findings contribute to understand the role of ADAM17 in oral cancer development and as a potential therapeutic target in oral cancer. In addition, our study also provides the basis for the development of novel and refined OSCC-targeting approaches.

Show MeSH
Related in: MedlinePlus