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Stanniocalcin-1 promotes tumor angiogenesis through up-regulation of VEGF in gastric cancer cells.

He LF, Wang TT, Gao QY, Zhao GF, Huang YH, Yu LK, Hou YY - J. Biomed. Sci. (2011)

Bottom Line: Inhibition of VEGF expression in supernatants with neutralizing antibody markedly abolished angiogenesis induced by STC-1 in vitro.The process of STC-1-regulated VEGF expression was mediated via PKCβII and ERK1/2.STC-1 promotes the expression of VEGF depended on the activation of PKCβII and ERK1/2 pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: Immunology and Reproductive Biology Lab, Medical School & State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, PR China.

ABSTRACT

Background: Stanniocalcin-1(STC-1) is up-regulated in several cancers including gastric cancer. Evidences suggest that STC-1 is associated with carcinogenesis and angiogenic process. However, it is unclear on the exact role for STC-1 in inducing angiogenesis and tumorigeneisis.

Method: BGC/STC cells (high-expression of STC-1) and BGC/shSTC cells (low- expression of STC-1) were constructed to investigate the effect of STC-1 on the xenograft tumor growth and angiogenesis in vitro and in vivo. ELISA assay was used to detect the expression of vascular endothelial growth factor (VEGF) in the supernatants. Neutralizing antibody was used to inhibit VEGF expression in supernatants. The expression of phosphorylated -PKCβII, phosphorylated -ERK1/2 and phosphorylated -P38 in the BGC treated with STC-1protein was detected by western blot.

Results: STC-1 could promote angiogenesis in vitro and in vivo, and the angiogenesis was consistent with VEGF expression in vitro. Inhibition of VEGF expression in supernatants with neutralizing antibody markedly abolished angiogenesis induced by STC-1 in vitro. The process of STC-1-regulated VEGF expression was mediated via PKCβII and ERK1/2.

Conclusions: STC-1 promotes the expression of VEGF depended on the activation of PKCβII and ERK1/2 pathways. VEGF subsequently enhances tumor angiogenesis which in turn promotes the gastric tumor growth.

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Effects of STC-1 and VEGF on HUVEC cell migration. (A) Effects of STC-1 on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with different conditioned supernatants. (B) Effects of VEGF on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with tumor supernatants incubated with 2 μg/mL VEGF monoclonal antibody (Bioactive). (C) The number of migration cells was quantified under a microscope at ×100 magnification. All histogram was carried out on multiple sections and the results are representative of three independent experiments. (D) Effect of isotype antibody on cell migration. IS: isotype antibody; V:VEGF neutralizing antibody; BGC/STC+IS: BGC/STC cell supernatants added with isotype antibody; BGC/STC+V: BGC/STC cell supernatants added with VEGF neutralizing antibody.
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Figure 3: Effects of STC-1 and VEGF on HUVEC cell migration. (A) Effects of STC-1 on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with different conditioned supernatants. (B) Effects of VEGF on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with tumor supernatants incubated with 2 μg/mL VEGF monoclonal antibody (Bioactive). (C) The number of migration cells was quantified under a microscope at ×100 magnification. All histogram was carried out on multiple sections and the results are representative of three independent experiments. (D) Effect of isotype antibody on cell migration. IS: isotype antibody; V:VEGF neutralizing antibody; BGC/STC+IS: BGC/STC cell supernatants added with isotype antibody; BGC/STC+V: BGC/STC cell supernatants added with VEGF neutralizing antibody.

Mentions: To determine the effect of STC-1 on angiogenesis, we use CFSE staining to detect proliferation rate of HUVECs. We found that BGC/STC culture supernatants could significantly promote HUVEC proliferation, while BGC/shSTC culture supernatants could inhibit HUVEC proliferation (Figure 2A). Then we considered whether the culture supernatants could regulate the migration of HUVEC. The Millicell cell culture insert was used to study the migration of the HUVEC in vitro. The migration of HUVEC was significantly enhanced with BGC/STC medium cultured, while the migration was reduced with BGC/shSTC medium cultured (Figure 2B, D). Tube formation assay was further verified the effect of STC-1 on this angiogenesis process. The formation of tube or cordlike structure could be induced by all kinds of tumor cell supernatants cultured with HUVEC, but not 1640 medium. Notably, BGC/STC supernatants showed an augmentation effect on the tube network while BGC/shSTC supernatants resulted in shorter and more blunted tubes (Figure 3A, C). These results suggest that STC-1 may change some factors of tumor microenvironment to modulate angiogenesis.


Stanniocalcin-1 promotes tumor angiogenesis through up-regulation of VEGF in gastric cancer cells.

He LF, Wang TT, Gao QY, Zhao GF, Huang YH, Yu LK, Hou YY - J. Biomed. Sci. (2011)

Effects of STC-1 and VEGF on HUVEC cell migration. (A) Effects of STC-1 on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with different conditioned supernatants. (B) Effects of VEGF on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with tumor supernatants incubated with 2 μg/mL VEGF monoclonal antibody (Bioactive). (C) The number of migration cells was quantified under a microscope at ×100 magnification. All histogram was carried out on multiple sections and the results are representative of three independent experiments. (D) Effect of isotype antibody on cell migration. IS: isotype antibody; V:VEGF neutralizing antibody; BGC/STC+IS: BGC/STC cell supernatants added with isotype antibody; BGC/STC+V: BGC/STC cell supernatants added with VEGF neutralizing antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3142497&req=5

Figure 3: Effects of STC-1 and VEGF on HUVEC cell migration. (A) Effects of STC-1 on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with different conditioned supernatants. (B) Effects of VEGF on HUVEC migration. HUVEC were seeded in triplicate on inserts, and incubated for 12 h with tumor supernatants incubated with 2 μg/mL VEGF monoclonal antibody (Bioactive). (C) The number of migration cells was quantified under a microscope at ×100 magnification. All histogram was carried out on multiple sections and the results are representative of three independent experiments. (D) Effect of isotype antibody on cell migration. IS: isotype antibody; V:VEGF neutralizing antibody; BGC/STC+IS: BGC/STC cell supernatants added with isotype antibody; BGC/STC+V: BGC/STC cell supernatants added with VEGF neutralizing antibody.
Mentions: To determine the effect of STC-1 on angiogenesis, we use CFSE staining to detect proliferation rate of HUVECs. We found that BGC/STC culture supernatants could significantly promote HUVEC proliferation, while BGC/shSTC culture supernatants could inhibit HUVEC proliferation (Figure 2A). Then we considered whether the culture supernatants could regulate the migration of HUVEC. The Millicell cell culture insert was used to study the migration of the HUVEC in vitro. The migration of HUVEC was significantly enhanced with BGC/STC medium cultured, while the migration was reduced with BGC/shSTC medium cultured (Figure 2B, D). Tube formation assay was further verified the effect of STC-1 on this angiogenesis process. The formation of tube or cordlike structure could be induced by all kinds of tumor cell supernatants cultured with HUVEC, but not 1640 medium. Notably, BGC/STC supernatants showed an augmentation effect on the tube network while BGC/shSTC supernatants resulted in shorter and more blunted tubes (Figure 3A, C). These results suggest that STC-1 may change some factors of tumor microenvironment to modulate angiogenesis.

Bottom Line: Inhibition of VEGF expression in supernatants with neutralizing antibody markedly abolished angiogenesis induced by STC-1 in vitro.The process of STC-1-regulated VEGF expression was mediated via PKCβII and ERK1/2.STC-1 promotes the expression of VEGF depended on the activation of PKCβII and ERK1/2 pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: Immunology and Reproductive Biology Lab, Medical School & State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, PR China.

ABSTRACT

Background: Stanniocalcin-1(STC-1) is up-regulated in several cancers including gastric cancer. Evidences suggest that STC-1 is associated with carcinogenesis and angiogenic process. However, it is unclear on the exact role for STC-1 in inducing angiogenesis and tumorigeneisis.

Method: BGC/STC cells (high-expression of STC-1) and BGC/shSTC cells (low- expression of STC-1) were constructed to investigate the effect of STC-1 on the xenograft tumor growth and angiogenesis in vitro and in vivo. ELISA assay was used to detect the expression of vascular endothelial growth factor (VEGF) in the supernatants. Neutralizing antibody was used to inhibit VEGF expression in supernatants. The expression of phosphorylated -PKCβII, phosphorylated -ERK1/2 and phosphorylated -P38 in the BGC treated with STC-1protein was detected by western blot.

Results: STC-1 could promote angiogenesis in vitro and in vivo, and the angiogenesis was consistent with VEGF expression in vitro. Inhibition of VEGF expression in supernatants with neutralizing antibody markedly abolished angiogenesis induced by STC-1 in vitro. The process of STC-1-regulated VEGF expression was mediated via PKCβII and ERK1/2.

Conclusions: STC-1 promotes the expression of VEGF depended on the activation of PKCβII and ERK1/2 pathways. VEGF subsequently enhances tumor angiogenesis which in turn promotes the gastric tumor growth.

Show MeSH
Related in: MedlinePlus