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α-santalol inhibits the angiogenesis and growth of human prostate tumor growth by targeting vascular endothelial growth factor receptor 2-mediated AKT/mTOR/P70S6K signaling pathway.

Saraswati S, Kumar S, Alhaider AA - Mol. Cancer (2013)

Bottom Line: However, recently, most of these anticancer drugs have some adverse effects.Discovery of novel VEGFR2 inhibitors as anticancer drug candidates is still needed.Furthermore, α-santalol reduced the cell viability and induced apoptosis in PC-3 cells, which were correlated with the downregulation of AKT, mTOR and P70S6K expressions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Camel Biomedical Research Unit, College of Pharmacy and Medicine, King Saud University, Riyadh, Kingdom of Saudi Arabia. saritasaraswati@gmail.com.

ABSTRACT

Background: VEGF receptor 2 (VEGFR2) inhibitors, as efficient antiangiogenesis agents, have been applied in the cancer treatment. However, recently, most of these anticancer drugs have some adverse effects. Discovery of novel VEGFR2 inhibitors as anticancer drug candidates is still needed.

Methods: We used α-santalol and analyzed its inhibitory effects on human umbilical vein endothelial cells (HUVECs) and Prostate tumor cells (PC-3 or LNCaP) in vitro. Tumor xenografts in nude mice were used to examine the in vivo activity of α-santalol.

Results: α-santalol significantly inhibits HUVEC proliferation, migration, invasion, and tube formation. Western blot analysis indicated that α-santalol inhibited VEGF-induced phosphorylation of VEGFR2 kinase and the downstream protein kinases including AKT, ERK, FAK, Src, mTOR, and pS6K in HUVEC, PC-3 and LNCaP cells. α-santalol treatment inhibited ex vivo and in vivo angiogenesis as evident by rat aortic and sponge implant angiogenesis assay. α-santalol significantly reduced the volume and the weight of solid tumors in prostate xenograft mouse model. The antiangiogenic effect by CD31 immunohistochemical staining indicated that α-santalol inhibited tumorigenesis by targeting angiogenesis. Furthermore, α-santalol reduced the cell viability and induced apoptosis in PC-3 cells, which were correlated with the downregulation of AKT, mTOR and P70S6K expressions. Molecular docking simulation indicated that α-santalol form hydrogen bonds and aromatic interactions within the ATP-binding region of the VEGFR2 kinase unit.

Conclusion: α-santalol inhibits angiogenesis by targeting VEGFR2 regulated AKT/mTOR/P70S6K signaling pathway, and could be used as a potential drug candidate for cancer therapy.

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α-santalol inhibits cell growth in both HUVEC and PC-3 cells. (A) MTT assays of HUVEC with α-santalol, vandetanib or sunitinib, respectively. (B) MTT assays of PC-3 cells with α-santalol, vandetanib or sunitinib, respectively. (C) α-santalol inhibits the VEGF induced proliferation of endothelial cells. (D) BrdU cell proliferation enzyme linked immunosorbent assay of HUVEC with α-santalol. (E) LDH cytotoxicity assay of HUVEC with α-santalol. Values are mean ± SEM (n = 6) of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 versus vehicle control. #p < 0.001 versus VEGF stimulated group.
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Figure 2: α-santalol inhibits cell growth in both HUVEC and PC-3 cells. (A) MTT assays of HUVEC with α-santalol, vandetanib or sunitinib, respectively. (B) MTT assays of PC-3 cells with α-santalol, vandetanib or sunitinib, respectively. (C) α-santalol inhibits the VEGF induced proliferation of endothelial cells. (D) BrdU cell proliferation enzyme linked immunosorbent assay of HUVEC with α-santalol. (E) LDH cytotoxicity assay of HUVEC with α-santalol. Values are mean ± SEM (n = 6) of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 versus vehicle control. #p < 0.001 versus VEGF stimulated group.

Mentions: Cell viability was determined by MTT assay as described previously [17,18]. At concentrations of 10–20 μM, α-santalol significantly inhibited endothelial cell proliferation with an IC50 value of 17.8 μM under normal culture conditions (Figure 2A). However, vandetanib and sunitinib inhibited cell viability at a much lower concentration with an IC50 value of 4.6 μM and 2.1 μM respectively (Figure 2A). α-santalol significantly (p < 0.01) inhibited PC-3 (Figure 2B) and LNCaP (Additional file 2: Figure S2) cell proliferation in the range of 20–40 μM as compared with the concentration of α-santalol required to suppress endothelial cell proliferation (10 μM), indicating that HUVECs were more sensitive to α-santalol than PC-3 (Figure 2A) or LNCaP (Additional file 2: Figure S2) cells induced inhibition in cell proliferation and promotion in cell apoptosis assays (Table 1). As angiogenesis is primarily initiated by growth factors, we next tested whether α-santalol decreased VEGF-mediated HUVEC proliferation and viability. We found that the α-santalol at 5 μM significantly inhibited VEGF-mediated HUVEC survival (p < 0.01) with an IC50 value of 10.16 μM (Figure 2C). As detected by BrdU incorporation assay (Figure 2D). DNA synthesis of HUVECs is also significantly inhibited by α-santalol (p < 0.05). To further examine whether α-santalol would result in toxic effects of HUVEC, LDH cytotoxic assay was carried out. α-santalol caused minute toxicity on HUVECs (Figure 2E).


α-santalol inhibits the angiogenesis and growth of human prostate tumor growth by targeting vascular endothelial growth factor receptor 2-mediated AKT/mTOR/P70S6K signaling pathway.

Saraswati S, Kumar S, Alhaider AA - Mol. Cancer (2013)

α-santalol inhibits cell growth in both HUVEC and PC-3 cells. (A) MTT assays of HUVEC with α-santalol, vandetanib or sunitinib, respectively. (B) MTT assays of PC-3 cells with α-santalol, vandetanib or sunitinib, respectively. (C) α-santalol inhibits the VEGF induced proliferation of endothelial cells. (D) BrdU cell proliferation enzyme linked immunosorbent assay of HUVEC with α-santalol. (E) LDH cytotoxicity assay of HUVEC with α-santalol. Values are mean ± SEM (n = 6) of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 versus vehicle control. #p < 0.001 versus VEGF stimulated group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4221991&req=5

Figure 2: α-santalol inhibits cell growth in both HUVEC and PC-3 cells. (A) MTT assays of HUVEC with α-santalol, vandetanib or sunitinib, respectively. (B) MTT assays of PC-3 cells with α-santalol, vandetanib or sunitinib, respectively. (C) α-santalol inhibits the VEGF induced proliferation of endothelial cells. (D) BrdU cell proliferation enzyme linked immunosorbent assay of HUVEC with α-santalol. (E) LDH cytotoxicity assay of HUVEC with α-santalol. Values are mean ± SEM (n = 6) of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 versus vehicle control. #p < 0.001 versus VEGF stimulated group.
Mentions: Cell viability was determined by MTT assay as described previously [17,18]. At concentrations of 10–20 μM, α-santalol significantly inhibited endothelial cell proliferation with an IC50 value of 17.8 μM under normal culture conditions (Figure 2A). However, vandetanib and sunitinib inhibited cell viability at a much lower concentration with an IC50 value of 4.6 μM and 2.1 μM respectively (Figure 2A). α-santalol significantly (p < 0.01) inhibited PC-3 (Figure 2B) and LNCaP (Additional file 2: Figure S2) cell proliferation in the range of 20–40 μM as compared with the concentration of α-santalol required to suppress endothelial cell proliferation (10 μM), indicating that HUVECs were more sensitive to α-santalol than PC-3 (Figure 2A) or LNCaP (Additional file 2: Figure S2) cells induced inhibition in cell proliferation and promotion in cell apoptosis assays (Table 1). As angiogenesis is primarily initiated by growth factors, we next tested whether α-santalol decreased VEGF-mediated HUVEC proliferation and viability. We found that the α-santalol at 5 μM significantly inhibited VEGF-mediated HUVEC survival (p < 0.01) with an IC50 value of 10.16 μM (Figure 2C). As detected by BrdU incorporation assay (Figure 2D). DNA synthesis of HUVECs is also significantly inhibited by α-santalol (p < 0.05). To further examine whether α-santalol would result in toxic effects of HUVEC, LDH cytotoxic assay was carried out. α-santalol caused minute toxicity on HUVECs (Figure 2E).

Bottom Line: However, recently, most of these anticancer drugs have some adverse effects.Discovery of novel VEGFR2 inhibitors as anticancer drug candidates is still needed.Furthermore, α-santalol reduced the cell viability and induced apoptosis in PC-3 cells, which were correlated with the downregulation of AKT, mTOR and P70S6K expressions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Camel Biomedical Research Unit, College of Pharmacy and Medicine, King Saud University, Riyadh, Kingdom of Saudi Arabia. saritasaraswati@gmail.com.

ABSTRACT

Background: VEGF receptor 2 (VEGFR2) inhibitors, as efficient antiangiogenesis agents, have been applied in the cancer treatment. However, recently, most of these anticancer drugs have some adverse effects. Discovery of novel VEGFR2 inhibitors as anticancer drug candidates is still needed.

Methods: We used α-santalol and analyzed its inhibitory effects on human umbilical vein endothelial cells (HUVECs) and Prostate tumor cells (PC-3 or LNCaP) in vitro. Tumor xenografts in nude mice were used to examine the in vivo activity of α-santalol.

Results: α-santalol significantly inhibits HUVEC proliferation, migration, invasion, and tube formation. Western blot analysis indicated that α-santalol inhibited VEGF-induced phosphorylation of VEGFR2 kinase and the downstream protein kinases including AKT, ERK, FAK, Src, mTOR, and pS6K in HUVEC, PC-3 and LNCaP cells. α-santalol treatment inhibited ex vivo and in vivo angiogenesis as evident by rat aortic and sponge implant angiogenesis assay. α-santalol significantly reduced the volume and the weight of solid tumors in prostate xenograft mouse model. The antiangiogenic effect by CD31 immunohistochemical staining indicated that α-santalol inhibited tumorigenesis by targeting angiogenesis. Furthermore, α-santalol reduced the cell viability and induced apoptosis in PC-3 cells, which were correlated with the downregulation of AKT, mTOR and P70S6K expressions. Molecular docking simulation indicated that α-santalol form hydrogen bonds and aromatic interactions within the ATP-binding region of the VEGFR2 kinase unit.

Conclusion: α-santalol inhibits angiogenesis by targeting VEGFR2 regulated AKT/mTOR/P70S6K signaling pathway, and could be used as a potential drug candidate for cancer therapy.

Show MeSH
Related in: MedlinePlus