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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

The propolis derivative, GS-002, induced ATF-3 expression that was mediated by MAPK pathways. Hep3B cells were treated with (a) 20 μg/mL GS-002 for the indicated time periods, or (b) with various concentrations of GS-002 for 1 h. Total cell lysates were used to detect protein expressions of p38, phosphor-p38 (p-p38), c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), extracellular signal-regulated kinase (ERK), and phospho-ERK (p-ERK) by Western blotting. Ten micrograms of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) was used as a positive control. (c) Hep3B cells were pretreated with 5 and 10 μM of the p38 inhibitor, SB203580, the ERK inhibitor, PD98059, or the JNK inhibitor, SP600125, for 90 min, then treated with GS-002 (20 μg/mL) for another 12 h, and the ATF-3 protein level was detected by Western blotting.
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fig6: The propolis derivative, GS-002, induced ATF-3 expression that was mediated by MAPK pathways. Hep3B cells were treated with (a) 20 μg/mL GS-002 for the indicated time periods, or (b) with various concentrations of GS-002 for 1 h. Total cell lysates were used to detect protein expressions of p38, phosphor-p38 (p-p38), c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), extracellular signal-regulated kinase (ERK), and phospho-ERK (p-ERK) by Western blotting. Ten micrograms of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) was used as a positive control. (c) Hep3B cells were pretreated with 5 and 10 μM of the p38 inhibitor, SB203580, the ERK inhibitor, PD98059, or the JNK inhibitor, SP600125, for 90 min, then treated with GS-002 (20 μg/mL) for another 12 h, and the ATF-3 protein level was detected by Western blotting.

Mentions: Activation of ATF-3 mainly depends on signaling pathways of MAPKs, which include ERK, JNK, and p38 kinase. To investigate whether GS-002 induced ATF-3 expression mediated by MAPK pathways, we examined phosphorylation levels of p38, JNK, and ERK in GS-002-treated cells. As shown in Figures 6(a) and 6(b), 20 μg/mL GS-002 markedly increased phosphorylation levels of p38, JNK, and ERK. To further demonstrate the importance of the activation of p38, ERK, and JNK in ATF-3 expression in GS-002-treated cells, SB203580, PD98059, and SP600125 were used to, respectively, inhibit the activities of p38, ERK, and JNK. As shown in Figure 6(c), SB203580, PD98059, and SP600125 markedly inhibited ATF-3 protein expression of GS-002-treated cells. The results suggest that ATF-3 expression is mainly mediated by activation of MAPK pathways in GS-002-treated cells.


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)

The propolis derivative, GS-002, induced ATF-3 expression that was mediated by MAPK pathways. Hep3B cells were treated with (a) 20 μg/mL GS-002 for the indicated time periods, or (b) with various concentrations of GS-002 for 1 h. Total cell lysates were used to detect protein expressions of p38, phosphor-p38 (p-p38), c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), extracellular signal-regulated kinase (ERK), and phospho-ERK (p-ERK) by Western blotting. Ten micrograms of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) was used as a positive control. (c) Hep3B cells were pretreated with 5 and 10 μM of the p38 inhibitor, SB203580, the ERK inhibitor, PD98059, or the JNK inhibitor, SP600125, for 90 min, then treated with GS-002 (20 μg/mL) for another 12 h, and the ATF-3 protein level was detected by Western blotting.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3814109&req=5

fig6: The propolis derivative, GS-002, induced ATF-3 expression that was mediated by MAPK pathways. Hep3B cells were treated with (a) 20 μg/mL GS-002 for the indicated time periods, or (b) with various concentrations of GS-002 for 1 h. Total cell lysates were used to detect protein expressions of p38, phosphor-p38 (p-p38), c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), extracellular signal-regulated kinase (ERK), and phospho-ERK (p-ERK) by Western blotting. Ten micrograms of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) was used as a positive control. (c) Hep3B cells were pretreated with 5 and 10 μM of the p38 inhibitor, SB203580, the ERK inhibitor, PD98059, or the JNK inhibitor, SP600125, for 90 min, then treated with GS-002 (20 μg/mL) for another 12 h, and the ATF-3 protein level was detected by Western blotting.
Mentions: Activation of ATF-3 mainly depends on signaling pathways of MAPKs, which include ERK, JNK, and p38 kinase. To investigate whether GS-002 induced ATF-3 expression mediated by MAPK pathways, we examined phosphorylation levels of p38, JNK, and ERK in GS-002-treated cells. As shown in Figures 6(a) and 6(b), 20 μg/mL GS-002 markedly increased phosphorylation levels of p38, JNK, and ERK. To further demonstrate the importance of the activation of p38, ERK, and JNK in ATF-3 expression in GS-002-treated cells, SB203580, PD98059, and SP600125 were used to, respectively, inhibit the activities of p38, ERK, and JNK. As shown in Figure 6(c), SB203580, PD98059, and SP600125 markedly inhibited ATF-3 protein expression of GS-002-treated cells. The results suggest that ATF-3 expression is mainly mediated by activation of MAPK pathways in GS-002-treated cells.

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