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The Ron receptor tyrosine kinase positively regulates angiogenic chemokine production in prostate cancer cells.

Thobe MN, Gurusamy D, Pathrose P, Waltz SE - Oncogene (2009)

Bottom Line: The knockdown of Ron in PC-3 or DU145 cells results in a significant decrease in angiogenic chemokine production and is associated with a decreased activation of the transcription factor nuclear factor-kappaB (NF-kappaB).Moreover, exogenous overexpression of Ron in LNCaP cells is sufficient to induce a significant increase in angiogenic chemokines that can be abrogated by inhibition of NF-kappaB signaling.Our data show that knockdown of Ron in prostate cancer cells results in significantly less endothelial cell chemotaxis when compared with Ron-expressing cells in vitro as well as in reduced tumor growth and decreased microvessel density after orthotopic transplantation into the prostate in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer and Cell Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.

ABSTRACT
Overexpression of the Ron receptor tyrosine kinase has recently been shown in a wide variety of human cancers. However, no studies have examined Ron receptor expression or function during prostate tumorigenesis. In this study we report that Ron is highly expressed in human prostate adenocarcinoma and metastatic lymph nodes when compared with normal prostate or benign prostate hyperplasia. Furthermore, we show that Ron is overexpressed in PC-3 and DU145 prostate cancer cell lines, and that the levels of angiogenic chemokines produced by prostate cancer cells positively correlate with Ron expression. The knockdown of Ron in PC-3 or DU145 cells results in a significant decrease in angiogenic chemokine production and is associated with a decreased activation of the transcription factor nuclear factor-kappaB (NF-kappaB). Moreover, exogenous overexpression of Ron in LNCaP cells is sufficient to induce a significant increase in angiogenic chemokines that can be abrogated by inhibition of NF-kappaB signaling. Given that the function of angiogenic chemokines is important in the development of new blood vessels, we also examined the ability of Ron to modulate endothelial cell migration. Our data show that knockdown of Ron in prostate cancer cells results in significantly less endothelial cell chemotaxis when compared with Ron-expressing cells in vitro as well as in reduced tumor growth and decreased microvessel density after orthotopic transplantation into the prostate in vivo. In total, our data suggest that the Ron receptor is important in modulating prostate tumor growth by modulating angiogenic chemokine production and subsequent endothelial cell recruitment.

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Ron receptor expression and activation in human prostate cellsA, Ron expression in human prostate cells. Ron is highly expressed in PC-3 and DU145 prostate cancer cells compared to 22RV1 and LNCaP cells as determined by Western analysis. B, and C, Ron is tyrosine phosphorylated and is an active kinase in DU145 and PC-3 cells but not in the non-transformed prostate cell line PZ-HPV-7. B, PZ-HPV-7, DU145 or PC-3 cells were immunoprecipitated with an antibody against Ron and were subjected to Western analysis with a phospho-tyrosine or a phosphorylated Ron (Tyrosine 1238/1239) antibody. A lysate control is shown representing 20% of the total input. C, DU145 or PC-3 cells were immunoprecipitated with a Ron antibody (or with IgG as a control), and a kinase assay was performed with myelin basic protein as substrate. Controls included reactions that did not include substrate.
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Figure 2: Ron receptor expression and activation in human prostate cellsA, Ron expression in human prostate cells. Ron is highly expressed in PC-3 and DU145 prostate cancer cells compared to 22RV1 and LNCaP cells as determined by Western analysis. B, and C, Ron is tyrosine phosphorylated and is an active kinase in DU145 and PC-3 cells but not in the non-transformed prostate cell line PZ-HPV-7. B, PZ-HPV-7, DU145 or PC-3 cells were immunoprecipitated with an antibody against Ron and were subjected to Western analysis with a phospho-tyrosine or a phosphorylated Ron (Tyrosine 1238/1239) antibody. A lysate control is shown representing 20% of the total input. C, DU145 or PC-3 cells were immunoprecipitated with a Ron antibody (or with IgG as a control), and a kinase assay was performed with myelin basic protein as substrate. Controls included reactions that did not include substrate.

Mentions: To determine the expression of Ron in human prostate cells, Western analyses were performed. As shown in Figure 2A, Western analysis demonstrates high Ron expression in PC-3 and DU145 cells and little Ron expression in 22RV1 and LNCaP cells. Interestingly, PC-3 and DU145 cells are two androgen-independent prostate cancer cell lines derived from metastatic prostate cancers. Further analysis of Ron expression (Supplemental Figure S1A) shows PC-3 cells express high amounts of Ron, while the non-invasive CA-HPV-10 cells derived from a primary human prostate tumor express an intermediate amount of Ron, and the non-transformed, immortalized prostate cell line PZ-HPV-7, has very little Ron expression. These results correlate with the pattern of Ron expression observed in the human prostate tumor tissue arrays shown in Figure 1 and Table 1.


The Ron receptor tyrosine kinase positively regulates angiogenic chemokine production in prostate cancer cells.

Thobe MN, Gurusamy D, Pathrose P, Waltz SE - Oncogene (2009)

Ron receptor expression and activation in human prostate cellsA, Ron expression in human prostate cells. Ron is highly expressed in PC-3 and DU145 prostate cancer cells compared to 22RV1 and LNCaP cells as determined by Western analysis. B, and C, Ron is tyrosine phosphorylated and is an active kinase in DU145 and PC-3 cells but not in the non-transformed prostate cell line PZ-HPV-7. B, PZ-HPV-7, DU145 or PC-3 cells were immunoprecipitated with an antibody against Ron and were subjected to Western analysis with a phospho-tyrosine or a phosphorylated Ron (Tyrosine 1238/1239) antibody. A lysate control is shown representing 20% of the total input. C, DU145 or PC-3 cells were immunoprecipitated with a Ron antibody (or with IgG as a control), and a kinase assay was performed with myelin basic protein as substrate. Controls included reactions that did not include substrate.
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Related In: Results  -  Collection

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Show All Figures
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Figure 2: Ron receptor expression and activation in human prostate cellsA, Ron expression in human prostate cells. Ron is highly expressed in PC-3 and DU145 prostate cancer cells compared to 22RV1 and LNCaP cells as determined by Western analysis. B, and C, Ron is tyrosine phosphorylated and is an active kinase in DU145 and PC-3 cells but not in the non-transformed prostate cell line PZ-HPV-7. B, PZ-HPV-7, DU145 or PC-3 cells were immunoprecipitated with an antibody against Ron and were subjected to Western analysis with a phospho-tyrosine or a phosphorylated Ron (Tyrosine 1238/1239) antibody. A lysate control is shown representing 20% of the total input. C, DU145 or PC-3 cells were immunoprecipitated with a Ron antibody (or with IgG as a control), and a kinase assay was performed with myelin basic protein as substrate. Controls included reactions that did not include substrate.
Mentions: To determine the expression of Ron in human prostate cells, Western analyses were performed. As shown in Figure 2A, Western analysis demonstrates high Ron expression in PC-3 and DU145 cells and little Ron expression in 22RV1 and LNCaP cells. Interestingly, PC-3 and DU145 cells are two androgen-independent prostate cancer cell lines derived from metastatic prostate cancers. Further analysis of Ron expression (Supplemental Figure S1A) shows PC-3 cells express high amounts of Ron, while the non-invasive CA-HPV-10 cells derived from a primary human prostate tumor express an intermediate amount of Ron, and the non-transformed, immortalized prostate cell line PZ-HPV-7, has very little Ron expression. These results correlate with the pattern of Ron expression observed in the human prostate tumor tissue arrays shown in Figure 1 and Table 1.

Bottom Line: The knockdown of Ron in PC-3 or DU145 cells results in a significant decrease in angiogenic chemokine production and is associated with a decreased activation of the transcription factor nuclear factor-kappaB (NF-kappaB).Moreover, exogenous overexpression of Ron in LNCaP cells is sufficient to induce a significant increase in angiogenic chemokines that can be abrogated by inhibition of NF-kappaB signaling.Our data show that knockdown of Ron in prostate cancer cells results in significantly less endothelial cell chemotaxis when compared with Ron-expressing cells in vitro as well as in reduced tumor growth and decreased microvessel density after orthotopic transplantation into the prostate in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer and Cell Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.

ABSTRACT
Overexpression of the Ron receptor tyrosine kinase has recently been shown in a wide variety of human cancers. However, no studies have examined Ron receptor expression or function during prostate tumorigenesis. In this study we report that Ron is highly expressed in human prostate adenocarcinoma and metastatic lymph nodes when compared with normal prostate or benign prostate hyperplasia. Furthermore, we show that Ron is overexpressed in PC-3 and DU145 prostate cancer cell lines, and that the levels of angiogenic chemokines produced by prostate cancer cells positively correlate with Ron expression. The knockdown of Ron in PC-3 or DU145 cells results in a significant decrease in angiogenic chemokine production and is associated with a decreased activation of the transcription factor nuclear factor-kappaB (NF-kappaB). Moreover, exogenous overexpression of Ron in LNCaP cells is sufficient to induce a significant increase in angiogenic chemokines that can be abrogated by inhibition of NF-kappaB signaling. Given that the function of angiogenic chemokines is important in the development of new blood vessels, we also examined the ability of Ron to modulate endothelial cell migration. Our data show that knockdown of Ron in prostate cancer cells results in significantly less endothelial cell chemotaxis when compared with Ron-expressing cells in vitro as well as in reduced tumor growth and decreased microvessel density after orthotopic transplantation into the prostate in vivo. In total, our data suggest that the Ron receptor is important in modulating prostate tumor growth by modulating angiogenic chemokine production and subsequent endothelial cell recruitment.

Show MeSH
Related in: MedlinePlus