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Non-caveolar caveolin-1 expression in prostate cancer cells promotes lymphangiogenesis.

Nassar ZD, Hill MM, Parton RG, Francois M, Parat MO - Oncoscience (2015)

Bottom Line: The effect of PCa cell conditioned media on lymphatic endothelial cell (LEC) viability, chemotaxis, chemokinesis and differentiation was assessed.In contrast, caveolar Cav-1 (in DU145 cells) did not significantly affect PCa cell lymphangiogenic potential.The effect of non-caveolar Cav-1 on LECs was mediated by increased expression of VEGF-A as demonstrated by neutralization by anti-VEGF-A antibody.

View Article: PubMed Central - PubMed

Affiliation: The University of Queensland, School of Pharmacy, QLD, Australia.

ABSTRACT
Lymphangiogenesis allows prostate cancer (PCa) lymphatic metastasis, which is associated with poor prognosis and short survival rates. Caveolin-1 (Cav-1) is a membrane protein localized in caveolae, but also exists in non-caveolar, cellular or extracellular forms. Cav-1 is overexpressed in PCa, promotes prostate tumour progression and metastasis. We investigated the effect of caveolar and non-caveolar Cav-1 on PCa lymphangiogenic potential. Cav-1 was down-regulated in PC3 and DU145, and ectopically expressed in LNCaP cells. The effect of PCa cell conditioned media on lymphatic endothelial cell (LEC) viability, chemotaxis, chemokinesis and differentiation was assessed. The effect of Cav-1 on PCa cell expression of lymphangiogenesis-modulators VEGF-A and VEGF-C was assessed using qPCR and ELISA of the conditioned medium. Non-caveolar Cav-1, whether exogenous or endogenous (in LNCaP and PC3 cells, respectively) enhanced LEC proliferation, migration and differentiation. In contrast, caveolar Cav-1 (in DU145 cells) did not significantly affect PCa cell lymphangiogenic potential. The effect of non-caveolar Cav-1 on LECs was mediated by increased expression of VEGF-A as demonstrated by neutralization by anti-VEGF-A antibody. This study unveils for the first time a crucial role for non-caveolar Cav-1 in modulating PCa cell expression of VEGF-A and subsequent LEC proliferation, migration and tube formation.

No MeSH data available.


Related in: MedlinePlus

Effect of Cav-1 expression in prostate cancer cells on LEC chemotaxis and chemokinesis(A) LEC ability to migrate randomly in two-dimension was tested using a wound healing assay during which cells were incubated in various prostate cancer cell-conditioned media for 6 h. Results are reported as percent of wound closure and shown as mean ± S.E.M. (n=3 separate experiments), *p<0.05. (B) LEC directional migration toward conditioned media from each prostate cancer cell line was assessed using the modified Boyden chamber assay. Results are reported as percent of the migration to conditioned medium of PCa control cells and shown as mean ± S.E.M. (n=3-5 separate experiments), **p<0.01.
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Figure 3: Effect of Cav-1 expression in prostate cancer cells on LEC chemotaxis and chemokinesis(A) LEC ability to migrate randomly in two-dimension was tested using a wound healing assay during which cells were incubated in various prostate cancer cell-conditioned media for 6 h. Results are reported as percent of wound closure and shown as mean ± S.E.M. (n=3 separate experiments), *p<0.05. (B) LEC directional migration toward conditioned media from each prostate cancer cell line was assessed using the modified Boyden chamber assay. Results are reported as percent of the migration to conditioned medium of PCa control cells and shown as mean ± S.E.M. (n=3-5 separate experiments), **p<0.01.

Mentions: We tested the ability of PCa cell-conditioned medium to promote chemokinesis by performing a scratch wound migration assay. LECs exposed to the conditioned medium of LNCaP cells expressing Cav-1 migrated significantly more than LECs exposed to CM of cells lacking Cav-1 (Figure 3a). In addition, the attenuation of Cav-1 expression in PC3 cells led to significantly decreased LEC migration, while there was no significant difference in the random migration rate between LECs exposed to conditioned medium of control DU145 versus Cav-1-down-regulated DU145 cells (Figure 3a). To evaluate the impact of Cav-1 expression in PCa cells on LEC chemotaxis, a modified Boyden chamber migration assay was conducted using PCa cell-conditioned media in the lower wells. The number of migrated LECs was significantly higher when using the medium of Cav-1-expressing LNCaP cells than when using the medium of LNCaP devoid of Cav-1. Similarly, knocking-down Cav-1 expression in PC3 decreased the production of LEC-attracting factor(s) compared with control PC3 cells. However, there was no statistically significant effect of Cav-1 down-regulation in DU145 cells on directed LEC migration (Figure 3b). These results suggest that Cav-1 expression in LNCaP and PC3, but not DU145, promotes LEC migration.


Non-caveolar caveolin-1 expression in prostate cancer cells promotes lymphangiogenesis.

Nassar ZD, Hill MM, Parton RG, Francois M, Parat MO - Oncoscience (2015)

Effect of Cav-1 expression in prostate cancer cells on LEC chemotaxis and chemokinesis(A) LEC ability to migrate randomly in two-dimension was tested using a wound healing assay during which cells were incubated in various prostate cancer cell-conditioned media for 6 h. Results are reported as percent of wound closure and shown as mean ± S.E.M. (n=3 separate experiments), *p<0.05. (B) LEC directional migration toward conditioned media from each prostate cancer cell line was assessed using the modified Boyden chamber assay. Results are reported as percent of the migration to conditioned medium of PCa control cells and shown as mean ± S.E.M. (n=3-5 separate experiments), **p<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549361&req=5

Figure 3: Effect of Cav-1 expression in prostate cancer cells on LEC chemotaxis and chemokinesis(A) LEC ability to migrate randomly in two-dimension was tested using a wound healing assay during which cells were incubated in various prostate cancer cell-conditioned media for 6 h. Results are reported as percent of wound closure and shown as mean ± S.E.M. (n=3 separate experiments), *p<0.05. (B) LEC directional migration toward conditioned media from each prostate cancer cell line was assessed using the modified Boyden chamber assay. Results are reported as percent of the migration to conditioned medium of PCa control cells and shown as mean ± S.E.M. (n=3-5 separate experiments), **p<0.01.
Mentions: We tested the ability of PCa cell-conditioned medium to promote chemokinesis by performing a scratch wound migration assay. LECs exposed to the conditioned medium of LNCaP cells expressing Cav-1 migrated significantly more than LECs exposed to CM of cells lacking Cav-1 (Figure 3a). In addition, the attenuation of Cav-1 expression in PC3 cells led to significantly decreased LEC migration, while there was no significant difference in the random migration rate between LECs exposed to conditioned medium of control DU145 versus Cav-1-down-regulated DU145 cells (Figure 3a). To evaluate the impact of Cav-1 expression in PCa cells on LEC chemotaxis, a modified Boyden chamber migration assay was conducted using PCa cell-conditioned media in the lower wells. The number of migrated LECs was significantly higher when using the medium of Cav-1-expressing LNCaP cells than when using the medium of LNCaP devoid of Cav-1. Similarly, knocking-down Cav-1 expression in PC3 decreased the production of LEC-attracting factor(s) compared with control PC3 cells. However, there was no statistically significant effect of Cav-1 down-regulation in DU145 cells on directed LEC migration (Figure 3b). These results suggest that Cav-1 expression in LNCaP and PC3, but not DU145, promotes LEC migration.

Bottom Line: The effect of PCa cell conditioned media on lymphatic endothelial cell (LEC) viability, chemotaxis, chemokinesis and differentiation was assessed.In contrast, caveolar Cav-1 (in DU145 cells) did not significantly affect PCa cell lymphangiogenic potential.The effect of non-caveolar Cav-1 on LECs was mediated by increased expression of VEGF-A as demonstrated by neutralization by anti-VEGF-A antibody.

View Article: PubMed Central - PubMed

Affiliation: The University of Queensland, School of Pharmacy, QLD, Australia.

ABSTRACT
Lymphangiogenesis allows prostate cancer (PCa) lymphatic metastasis, which is associated with poor prognosis and short survival rates. Caveolin-1 (Cav-1) is a membrane protein localized in caveolae, but also exists in non-caveolar, cellular or extracellular forms. Cav-1 is overexpressed in PCa, promotes prostate tumour progression and metastasis. We investigated the effect of caveolar and non-caveolar Cav-1 on PCa lymphangiogenic potential. Cav-1 was down-regulated in PC3 and DU145, and ectopically expressed in LNCaP cells. The effect of PCa cell conditioned media on lymphatic endothelial cell (LEC) viability, chemotaxis, chemokinesis and differentiation was assessed. The effect of Cav-1 on PCa cell expression of lymphangiogenesis-modulators VEGF-A and VEGF-C was assessed using qPCR and ELISA of the conditioned medium. Non-caveolar Cav-1, whether exogenous or endogenous (in LNCaP and PC3 cells, respectively) enhanced LEC proliferation, migration and differentiation. In contrast, caveolar Cav-1 (in DU145 cells) did not significantly affect PCa cell lymphangiogenic potential. The effect of non-caveolar Cav-1 on LECs was mediated by increased expression of VEGF-A as demonstrated by neutralization by anti-VEGF-A antibody. This study unveils for the first time a crucial role for non-caveolar Cav-1 in modulating PCa cell expression of VEGF-A and subsequent LEC proliferation, migration and tube formation.

No MeSH data available.


Related in: MedlinePlus