Limits...
A Taiwanese Propolis Derivative Induces Apoptosis through Inducing Endoplasmic Reticular Stress and Activating Transcription Factor-3 in Human Hepatoma Cells.

Suk FM, Lien GS, Huang WJ, Chen CN, Lu SY, Yang YC, Yan MD, Liang YC - Evid Based Complement Alternat Med (2013)

Bottom Line: First, we found that GS-002 significantly inhibited cell proliferation and induced cell apoptosis in dose-dependent manners.Furthermore, we found that GS-002 induced more cell apoptosis in ATF-3-overexpressing cells.These results suggest that the induction of apoptosis by the propolis derivative, GS-002, is partially mediated through ER stress and ATF-3-dependent pathways, and GS-002 has the potential for development as an antitumor drug.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastroenterology, Department of Internal Medicine, Wan Fang Hospital, Taipei Medical University, Taipei 116, Taiwan.

ABSTRACT
Activating transcription factor-(ATF-) 3, a stress-inducible transcription factor, is rapidly upregulated under various stress conditions and plays an important role in inducing cancer cell apoptosis. NBM-TP-007-GS-002 (GS-002) is a Taiwanese propolin G (PPG) derivative. In this study, we examined the antitumor effects of GS-002 in human hepatoma Hep3B and HepG2 cells in vitro. First, we found that GS-002 significantly inhibited cell proliferation and induced cell apoptosis in dose-dependent manners. Several main apoptotic indicators were found in GS-002-treated cells, such as the cleaved forms of caspase-3, caspase-9, and poly(ADP-ribose) polymerase (PARP). GS-002 also induced endoplasmic reticular (ER) stress as evidenced by increases in ER stress-responsive proteins including glucose-regulated protein 78 (GRP78), growth arrest- and DNA damage-inducible gene 153 (GADD153), phosphorylated eukaryotic initiation factor 2 α (eIF2 α ), phosphorylated protein endoplasmic-reticular-resident kinase (PERK), and ATF-3. The induction of ATF-3 expression was mediated by mitogen-activated protein kinase (MAPK) signaling pathways in GS-002-treated cells. Furthermore, we found that GS-002 induced more cell apoptosis in ATF-3-overexpressing cells. These results suggest that the induction of apoptosis by the propolis derivative, GS-002, is partially mediated through ER stress and ATF-3-dependent pathways, and GS-002 has the potential for development as an antitumor drug.

No MeSH data available.


Related in: MedlinePlus

Overexpression of ATF-3 resulted in enhancement of cell apoptosis in human hepatoma cells treated with the propolis derivative, GS-002. (a) Hep3B cells were transfected with various doses of the pCI-ATF3 plasmid for 24 h, and total cell lysates were used to detect the ATF-3 protein level by Western blotting. (b) and (c) Hep3B cells were transfected with 2 μg of the pCI-ATF3 plasmid for 24 h and then treated with GS-002 for another 24 h. (b) The viable cell number was determined by a crystal violet dye. Values are presented as the mean ± SE of triplcate tests. *P < 0.05 versus individual pcDNA3-transfected control cells. (c) Total cell lysates were used to detect protein expression of the full-length PPAR (FU-PPAR), cleaved form of PPAR (CF-PPAR), cleaved form of caspase-3 (CF-caspase-3), cleaved form of caspase-9 (CF-caspase-9), ATF-3, and α-tubulin by Western blotting.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3814109&req=5

fig7: Overexpression of ATF-3 resulted in enhancement of cell apoptosis in human hepatoma cells treated with the propolis derivative, GS-002. (a) Hep3B cells were transfected with various doses of the pCI-ATF3 plasmid for 24 h, and total cell lysates were used to detect the ATF-3 protein level by Western blotting. (b) and (c) Hep3B cells were transfected with 2 μg of the pCI-ATF3 plasmid for 24 h and then treated with GS-002 for another 24 h. (b) The viable cell number was determined by a crystal violet dye. Values are presented as the mean ± SE of triplcate tests. *P < 0.05 versus individual pcDNA3-transfected control cells. (c) Total cell lysates were used to detect protein expression of the full-length PPAR (FU-PPAR), cleaved form of PPAR (CF-PPAR), cleaved form of caspase-3 (CF-caspase-3), cleaved form of caspase-9 (CF-caspase-9), ATF-3, and α-tubulin by Western blotting.

Mentions: To understand the role of ATF-3 in GS-002-induced apoptosis in hepatoma cells, Hep3B cells were transitionally transfected with the ATF-3 expression plasmid, pCI-ATF3. As shown in Figure 7(a), transfection with >2 μg of pCI-ATF3 plasmid significantly increased ATF-3 protein expression. Induction of apoptosis was significantly enhanced by transfection with the pCI-ATF3 plasmid at various doses of GS-002 (Figure 7(b)), indicating that GS-002 induced greater cell apoptosis in ATF-3-overexpressing cells than in control cells. The cleaved forms of PARP and caspase-3 also increased in ATF-3-overexpressing cells (Figure 7(c)). These results suggest that induction of apoptosis by GS-002 is mediated through an ATF-3-dependent pathway.


A Taiwanese Propolis Derivative Induces Apoptosis through Inducing Endoplasmic Reticular Stress and Activating Transcription Factor-3 in Human Hepatoma Cells.

Suk FM, Lien GS, Huang WJ, Chen CN, Lu SY, Yang YC, Yan MD, Liang YC - Evid Based Complement Alternat Med (2013)

Overexpression of ATF-3 resulted in enhancement of cell apoptosis in human hepatoma cells treated with the propolis derivative, GS-002. (a) Hep3B cells were transfected with various doses of the pCI-ATF3 plasmid for 24 h, and total cell lysates were used to detect the ATF-3 protein level by Western blotting. (b) and (c) Hep3B cells were transfected with 2 μg of the pCI-ATF3 plasmid for 24 h and then treated with GS-002 for another 24 h. (b) The viable cell number was determined by a crystal violet dye. Values are presented as the mean ± SE of triplcate tests. *P < 0.05 versus individual pcDNA3-transfected control cells. (c) Total cell lysates were used to detect protein expression of the full-length PPAR (FU-PPAR), cleaved form of PPAR (CF-PPAR), cleaved form of caspase-3 (CF-caspase-3), cleaved form of caspase-9 (CF-caspase-9), ATF-3, and α-tubulin by Western blotting.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3814109&req=5

fig7: Overexpression of ATF-3 resulted in enhancement of cell apoptosis in human hepatoma cells treated with the propolis derivative, GS-002. (a) Hep3B cells were transfected with various doses of the pCI-ATF3 plasmid for 24 h, and total cell lysates were used to detect the ATF-3 protein level by Western blotting. (b) and (c) Hep3B cells were transfected with 2 μg of the pCI-ATF3 plasmid for 24 h and then treated with GS-002 for another 24 h. (b) The viable cell number was determined by a crystal violet dye. Values are presented as the mean ± SE of triplcate tests. *P < 0.05 versus individual pcDNA3-transfected control cells. (c) Total cell lysates were used to detect protein expression of the full-length PPAR (FU-PPAR), cleaved form of PPAR (CF-PPAR), cleaved form of caspase-3 (CF-caspase-3), cleaved form of caspase-9 (CF-caspase-9), ATF-3, and α-tubulin by Western blotting.
Mentions: To understand the role of ATF-3 in GS-002-induced apoptosis in hepatoma cells, Hep3B cells were transitionally transfected with the ATF-3 expression plasmid, pCI-ATF3. As shown in Figure 7(a), transfection with >2 μg of pCI-ATF3 plasmid significantly increased ATF-3 protein expression. Induction of apoptosis was significantly enhanced by transfection with the pCI-ATF3 plasmid at various doses of GS-002 (Figure 7(b)), indicating that GS-002 induced greater cell apoptosis in ATF-3-overexpressing cells than in control cells. The cleaved forms of PARP and caspase-3 also increased in ATF-3-overexpressing cells (Figure 7(c)). These results suggest that induction of apoptosis by GS-002 is mediated through an ATF-3-dependent pathway.

Bottom Line: First, we found that GS-002 significantly inhibited cell proliferation and induced cell apoptosis in dose-dependent manners.Furthermore, we found that GS-002 induced more cell apoptosis in ATF-3-overexpressing cells.These results suggest that the induction of apoptosis by the propolis derivative, GS-002, is partially mediated through ER stress and ATF-3-dependent pathways, and GS-002 has the potential for development as an antitumor drug.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastroenterology, Department of Internal Medicine, Wan Fang Hospital, Taipei Medical University, Taipei 116, Taiwan.

ABSTRACT
Activating transcription factor-(ATF-) 3, a stress-inducible transcription factor, is rapidly upregulated under various stress conditions and plays an important role in inducing cancer cell apoptosis. NBM-TP-007-GS-002 (GS-002) is a Taiwanese propolin G (PPG) derivative. In this study, we examined the antitumor effects of GS-002 in human hepatoma Hep3B and HepG2 cells in vitro. First, we found that GS-002 significantly inhibited cell proliferation and induced cell apoptosis in dose-dependent manners. Several main apoptotic indicators were found in GS-002-treated cells, such as the cleaved forms of caspase-3, caspase-9, and poly(ADP-ribose) polymerase (PARP). GS-002 also induced endoplasmic reticular (ER) stress as evidenced by increases in ER stress-responsive proteins including glucose-regulated protein 78 (GRP78), growth arrest- and DNA damage-inducible gene 153 (GADD153), phosphorylated eukaryotic initiation factor 2 α (eIF2 α ), phosphorylated protein endoplasmic-reticular-resident kinase (PERK), and ATF-3. The induction of ATF-3 expression was mediated by mitogen-activated protein kinase (MAPK) signaling pathways in GS-002-treated cells. Furthermore, we found that GS-002 induced more cell apoptosis in ATF-3-overexpressing cells. These results suggest that the induction of apoptosis by the propolis derivative, GS-002, is partially mediated through ER stress and ATF-3-dependent pathways, and GS-002 has the potential for development as an antitumor drug.

No MeSH data available.


Related in: MedlinePlus