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VEGF-A/NRP1 stimulates GIPC1 and Syx complex formation to promote RhoA activation and proliferation in skin cancer cells.

Yoshida A, Shimizu A, Asano H, Kadonosono T, Kondoh SK, Geretti E, Mammoto A, Klagsbrun M, Seo MK - Biol Open (2015)

Bottom Line: DJM-1, a human skin cancer cell line, expresses endogenous VEGF-A and NRP1.Furthermore, the inhibition of VEGF-A/NRP1 signaling upregulated p27, a CDK inhibitor.In conclusion, this new signaling pathway of VEGF-A/NRP1 induced cancer cell proliferation by forming a GIPC1/Syx complex that activated RhoA to degrade the p27 protein.

View Article: PubMed Central - PubMed

Affiliation: Division of Engineering (Biotechnology), Graduate School of Engineering, Kyoto Sangyo University, Kyoto 603-8555, Japan.

No MeSH data available.


Related in: MedlinePlus

The VEGFR kinase inhibitor SU5614 and Avastin did not inhibit DJM-1 cell proliferation. (A) Western blot for VEGFR1 or VEGFR2 of DJM-1 cell lysates. As a positive control, the cell lysates of HUVEC were applied in the left lanes. Arrows indicate VEGFR1 or VEGFR2. (B) Colony formation assay for DJM-1 cells treated with 10 µM SU5614, the VEGFR kinase inhibitor, and with 0.2% DMSO as the control. (C) DJM-1 cell colony formation assay treated with Avastin (from 1 to 250 µg/ml). These data represent the means±s.d. N.S., not significant. Percentages from each mean relative to the DMSO (B) or no addtion (C) are shown below the graph.
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BIO010918F2: The VEGFR kinase inhibitor SU5614 and Avastin did not inhibit DJM-1 cell proliferation. (A) Western blot for VEGFR1 or VEGFR2 of DJM-1 cell lysates. As a positive control, the cell lysates of HUVEC were applied in the left lanes. Arrows indicate VEGFR1 or VEGFR2. (B) Colony formation assay for DJM-1 cells treated with 10 µM SU5614, the VEGFR kinase inhibitor, and with 0.2% DMSO as the control. (C) DJM-1 cell colony formation assay treated with Avastin (from 1 to 250 µg/ml). These data represent the means±s.d. N.S., not significant. Percentages from each mean relative to the DMSO (B) or no addtion (C) are shown below the graph.

Mentions: VEGF-A has multiple receptors: VEGFR1, VEGFR2, and neuropilins 1 and 2 (Ferrara, 2009). The expression of VEGFR1 and VEGFR2 was detected by western blotting in HUVEC, but not in DJM-1 cells (Fig. 2A). In order to determine whether VEGFR1 or VEGFR2 signaling occurred in DJM-1 cells in response to VEGF-A, the effects of SU5614, a VEGFR tyrosine kinase inhibitor, were examined in DJM-1 cells in soft agar (Fig. 2B). However, SU5614 did not inhibit the proliferation of DJM-1 cells (DMSO: 100%, SU5614: 96%). Avastin is an antibody that neutralizes VEGF-A and targets VEGFR-binding sites. However, Avastin did not inhibit DJM-1 cell proliferation (no addition: 100%, 1 µg/ml: 98%, 10 µg/ml: 96%, 250 µg/ml: 94%, respectively) (Fig. 2C). These results suggested that autocrine VEGF-A induced cancer proliferation, but did not mediate the VEGFR1 or VEGFR2 signaling pathway.Fig. 2.


VEGF-A/NRP1 stimulates GIPC1 and Syx complex formation to promote RhoA activation and proliferation in skin cancer cells.

Yoshida A, Shimizu A, Asano H, Kadonosono T, Kondoh SK, Geretti E, Mammoto A, Klagsbrun M, Seo MK - Biol Open (2015)

The VEGFR kinase inhibitor SU5614 and Avastin did not inhibit DJM-1 cell proliferation. (A) Western blot for VEGFR1 or VEGFR2 of DJM-1 cell lysates. As a positive control, the cell lysates of HUVEC were applied in the left lanes. Arrows indicate VEGFR1 or VEGFR2. (B) Colony formation assay for DJM-1 cells treated with 10 µM SU5614, the VEGFR kinase inhibitor, and with 0.2% DMSO as the control. (C) DJM-1 cell colony formation assay treated with Avastin (from 1 to 250 µg/ml). These data represent the means±s.d. N.S., not significant. Percentages from each mean relative to the DMSO (B) or no addtion (C) are shown below the graph.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582117&req=5

BIO010918F2: The VEGFR kinase inhibitor SU5614 and Avastin did not inhibit DJM-1 cell proliferation. (A) Western blot for VEGFR1 or VEGFR2 of DJM-1 cell lysates. As a positive control, the cell lysates of HUVEC were applied in the left lanes. Arrows indicate VEGFR1 or VEGFR2. (B) Colony formation assay for DJM-1 cells treated with 10 µM SU5614, the VEGFR kinase inhibitor, and with 0.2% DMSO as the control. (C) DJM-1 cell colony formation assay treated with Avastin (from 1 to 250 µg/ml). These data represent the means±s.d. N.S., not significant. Percentages from each mean relative to the DMSO (B) or no addtion (C) are shown below the graph.
Mentions: VEGF-A has multiple receptors: VEGFR1, VEGFR2, and neuropilins 1 and 2 (Ferrara, 2009). The expression of VEGFR1 and VEGFR2 was detected by western blotting in HUVEC, but not in DJM-1 cells (Fig. 2A). In order to determine whether VEGFR1 or VEGFR2 signaling occurred in DJM-1 cells in response to VEGF-A, the effects of SU5614, a VEGFR tyrosine kinase inhibitor, were examined in DJM-1 cells in soft agar (Fig. 2B). However, SU5614 did not inhibit the proliferation of DJM-1 cells (DMSO: 100%, SU5614: 96%). Avastin is an antibody that neutralizes VEGF-A and targets VEGFR-binding sites. However, Avastin did not inhibit DJM-1 cell proliferation (no addition: 100%, 1 µg/ml: 98%, 10 µg/ml: 96%, 250 µg/ml: 94%, respectively) (Fig. 2C). These results suggested that autocrine VEGF-A induced cancer proliferation, but did not mediate the VEGFR1 or VEGFR2 signaling pathway.Fig. 2.

Bottom Line: DJM-1, a human skin cancer cell line, expresses endogenous VEGF-A and NRP1.Furthermore, the inhibition of VEGF-A/NRP1 signaling upregulated p27, a CDK inhibitor.In conclusion, this new signaling pathway of VEGF-A/NRP1 induced cancer cell proliferation by forming a GIPC1/Syx complex that activated RhoA to degrade the p27 protein.

View Article: PubMed Central - PubMed

Affiliation: Division of Engineering (Biotechnology), Graduate School of Engineering, Kyoto Sangyo University, Kyoto 603-8555, Japan.

No MeSH data available.


Related in: MedlinePlus