Limits...
Isoindolone derivative QSN-10c induces leukemic cell apoptosis and suppresses angiogenesis via PI3K/AKT signaling pathway inhibition.

Lv WW, Qin SN, Chen CQ, Zhang JJ, Ren TS, Xu YN, Zhao QC - Acta Pharmacol. Sin. (2014)

Bottom Line: Furthermore, QSN-10c dose-dependently decreased the Δψm in K562 cells, increased the release of cytochrome c and the level of Bax, and decreased the level of Bcl-2, suggesting that QSN-10c-induced apoptosis of K562 cells was mediated via the mitochondrial apoptotic pathway.QSN-10c treatment did not alter the Δψm in HUVECs, but dose-dependently inhibited the expression of VEGF, inhibited the tube formation and cell migration in vitro, and significantly suppressed the number of ISVs in zebrafish embryos in vivo.QSN-10c is a novel antitumor compound that exerts both antitumor and anti-angiogenic effects via inhibiting the PI3K/AKT/GSK3β signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China [2] Shenyang Pharmaceutical University, Shenyang 110016, China.

ABSTRACT

Aim: 2-(4,6-Dimethoxy-1,3-dioxoisoindolin-2-yl) ethyl 2-chloroacetate (QSN-10c) is one of isoindolone derivatives with antiproliferative activity against human umbilical vein endothelial cells (HUVECs). The aim of this study was to investigate its antitumor activity in vitro and anti-angiogenic effects in vitro and in vivo.

Methods: K562 leukemic cells and HUVECs were used for in vitro studies. Cell viability was examined using MTT assay. Cell apoptosis and mitochondrial transmembrane potential (Δψm) were detected with flow cytometry. Tube formation and migration of HUVECs were studied using two-dimensional Matrigel assay and wound-healing migration assay, respectively. VEGF levels were analyzed with RT-PCR and Western blotting. A zebrafish embryo model was used for in vivo anti-angiogenic studies. The molecular mechanisms for apoptosis in K562 cells and antiangiogenesis were measured with Western blotting.

Results: In antitumor activity studies, QSN-10c suppressed the viability of K562 cells and induced apoptosis in dose- and time-dependent manners. Furthermore, QSN-10c dose-dependently decreased the Δψm in K562 cells, increased the release of cytochrome c and the level of Bax, and decreased the level of Bcl-2, suggesting that QSN-10c-induced apoptosis of K562 cells was mediated via the mitochondrial apoptotic pathway. In anti-angiogenic activity studies, QSN-10c suppressed the viability of HUVECs and induced apoptosis in dose dependent manners. QSN-10c treatment did not alter the Δψm in HUVECs, but dose-dependently inhibited the expression of VEGF, inhibited the tube formation and cell migration in vitro, and significantly suppressed the number of ISVs in zebrafish embryos in vivo. Furthermore, QSN-10c dose-dependently suppressed the phosphorylation of AKT and GSK3β in both HUVECs and K562 cells.

Conclusion: QSN-10c is a novel antitumor compound that exerts both antitumor and anti-angiogenic effects via inhibiting the PI3K/AKT/GSK3β signaling pathway.

Show MeSH
QSN-10c inhibits capillary structure formation of endothelial cells on Matrigel. (A) HUVECs cultured on Matrigel were treated with only DMSO (0.1%). (B, C) HUVECs cultured on Matrigel were treated with 100 μmol/L of thalidomide and 4,6-dihydroxyisoindol-1,3-dione, respectively. (D–F) HUVECs cultured on Matrigel were treated with various concentrations of QSN-10c (50, 100, and 200 μmol/L). (G) Quantitative comparison of the numbers of branching points in different groups. Cells receiving only DMSO (0.1%) served as a vehicle control, thalidomide as a positive control. Data are expressed as percentages of the vehicle control (100%) in mean±SD from three independent experiments. bP<0.05, cP<0.01 compared with control.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4814032&req=5

fig5: QSN-10c inhibits capillary structure formation of endothelial cells on Matrigel. (A) HUVECs cultured on Matrigel were treated with only DMSO (0.1%). (B, C) HUVECs cultured on Matrigel were treated with 100 μmol/L of thalidomide and 4,6-dihydroxyisoindol-1,3-dione, respectively. (D–F) HUVECs cultured on Matrigel were treated with various concentrations of QSN-10c (50, 100, and 200 μmol/L). (G) Quantitative comparison of the numbers of branching points in different groups. Cells receiving only DMSO (0.1%) served as a vehicle control, thalidomide as a positive control. Data are expressed as percentages of the vehicle control (100%) in mean±SD from three independent experiments. bP<0.05, cP<0.01 compared with control.

Mentions: Although angiogenesis is a complex procedure involving various cell types, endothelial cell tube formation is a key step of angiogenesis32. To further assess the effects of QSN-10c on endothelial cell tube formation, we used a two-dimensional Matrigel assay to examine the effect of QSN-10c on HUVEC tube formation. When HUVECs were seeded on the Matrigel, robust tubular-like structures were formed. Capillary tube formation on Matrigel was inhibited by approximately 7.4% after treatment with 25 μmol/L QSN-10c for 8 h (P>0.05). Tube formation was inhibited by approximately 50% with 50 μmol/L QSN-10c, and tube formation was almost completely inhibited upon treatment with 200 μmol/L. The QSN-10c-mediated effects were more robust than the results of treatment with 4,6-dihydroxyisoindol-1,3-dione and thalidomide (Figure 5A–5G, P<0.05, P<0.01).


Isoindolone derivative QSN-10c induces leukemic cell apoptosis and suppresses angiogenesis via PI3K/AKT signaling pathway inhibition.

Lv WW, Qin SN, Chen CQ, Zhang JJ, Ren TS, Xu YN, Zhao QC - Acta Pharmacol. Sin. (2014)

QSN-10c inhibits capillary structure formation of endothelial cells on Matrigel. (A) HUVECs cultured on Matrigel were treated with only DMSO (0.1%). (B, C) HUVECs cultured on Matrigel were treated with 100 μmol/L of thalidomide and 4,6-dihydroxyisoindol-1,3-dione, respectively. (D–F) HUVECs cultured on Matrigel were treated with various concentrations of QSN-10c (50, 100, and 200 μmol/L). (G) Quantitative comparison of the numbers of branching points in different groups. Cells receiving only DMSO (0.1%) served as a vehicle control, thalidomide as a positive control. Data are expressed as percentages of the vehicle control (100%) in mean±SD from three independent experiments. bP<0.05, cP<0.01 compared with control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4814032&req=5

fig5: QSN-10c inhibits capillary structure formation of endothelial cells on Matrigel. (A) HUVECs cultured on Matrigel were treated with only DMSO (0.1%). (B, C) HUVECs cultured on Matrigel were treated with 100 μmol/L of thalidomide and 4,6-dihydroxyisoindol-1,3-dione, respectively. (D–F) HUVECs cultured on Matrigel were treated with various concentrations of QSN-10c (50, 100, and 200 μmol/L). (G) Quantitative comparison of the numbers of branching points in different groups. Cells receiving only DMSO (0.1%) served as a vehicle control, thalidomide as a positive control. Data are expressed as percentages of the vehicle control (100%) in mean±SD from three independent experiments. bP<0.05, cP<0.01 compared with control.
Mentions: Although angiogenesis is a complex procedure involving various cell types, endothelial cell tube formation is a key step of angiogenesis32. To further assess the effects of QSN-10c on endothelial cell tube formation, we used a two-dimensional Matrigel assay to examine the effect of QSN-10c on HUVEC tube formation. When HUVECs were seeded on the Matrigel, robust tubular-like structures were formed. Capillary tube formation on Matrigel was inhibited by approximately 7.4% after treatment with 25 μmol/L QSN-10c for 8 h (P>0.05). Tube formation was inhibited by approximately 50% with 50 μmol/L QSN-10c, and tube formation was almost completely inhibited upon treatment with 200 μmol/L. The QSN-10c-mediated effects were more robust than the results of treatment with 4,6-dihydroxyisoindol-1,3-dione and thalidomide (Figure 5A–5G, P<0.05, P<0.01).

Bottom Line: Furthermore, QSN-10c dose-dependently decreased the Δψm in K562 cells, increased the release of cytochrome c and the level of Bax, and decreased the level of Bcl-2, suggesting that QSN-10c-induced apoptosis of K562 cells was mediated via the mitochondrial apoptotic pathway.QSN-10c treatment did not alter the Δψm in HUVECs, but dose-dependently inhibited the expression of VEGF, inhibited the tube formation and cell migration in vitro, and significantly suppressed the number of ISVs in zebrafish embryos in vivo.QSN-10c is a novel antitumor compound that exerts both antitumor and anti-angiogenic effects via inhibiting the PI3K/AKT/GSK3β signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China [2] Shenyang Pharmaceutical University, Shenyang 110016, China.

ABSTRACT

Aim: 2-(4,6-Dimethoxy-1,3-dioxoisoindolin-2-yl) ethyl 2-chloroacetate (QSN-10c) is one of isoindolone derivatives with antiproliferative activity against human umbilical vein endothelial cells (HUVECs). The aim of this study was to investigate its antitumor activity in vitro and anti-angiogenic effects in vitro and in vivo.

Methods: K562 leukemic cells and HUVECs were used for in vitro studies. Cell viability was examined using MTT assay. Cell apoptosis and mitochondrial transmembrane potential (Δψm) were detected with flow cytometry. Tube formation and migration of HUVECs were studied using two-dimensional Matrigel assay and wound-healing migration assay, respectively. VEGF levels were analyzed with RT-PCR and Western blotting. A zebrafish embryo model was used for in vivo anti-angiogenic studies. The molecular mechanisms for apoptosis in K562 cells and antiangiogenesis were measured with Western blotting.

Results: In antitumor activity studies, QSN-10c suppressed the viability of K562 cells and induced apoptosis in dose- and time-dependent manners. Furthermore, QSN-10c dose-dependently decreased the Δψm in K562 cells, increased the release of cytochrome c and the level of Bax, and decreased the level of Bcl-2, suggesting that QSN-10c-induced apoptosis of K562 cells was mediated via the mitochondrial apoptotic pathway. In anti-angiogenic activity studies, QSN-10c suppressed the viability of HUVECs and induced apoptosis in dose dependent manners. QSN-10c treatment did not alter the Δψm in HUVECs, but dose-dependently inhibited the expression of VEGF, inhibited the tube formation and cell migration in vitro, and significantly suppressed the number of ISVs in zebrafish embryos in vivo. Furthermore, QSN-10c dose-dependently suppressed the phosphorylation of AKT and GSK3β in both HUVECs and K562 cells.

Conclusion: QSN-10c is a novel antitumor compound that exerts both antitumor and anti-angiogenic effects via inhibiting the PI3K/AKT/GSK3β signaling pathway.

Show MeSH