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Fatsioside A‑induced apoptotic death of HepG2 cells requires activation of AMP‑activated protein kinase.

Zheng YS, Zhang JY, Zhang DH - Mol Med Rep (2015)

Bottom Line: The results of the present study demonstrated that fatsioside A induced apoptotic death of the human HepG2 HCC cells, which was associated with a marked activation of AMPK and increased expression of the downstream acetyl‑CoA carboxylase carboxylase.Inhibition of AMPK by RNA interference or by its inhibitor, compound C, suppressed fatsioside A‑induced caspase‑3 cleavage and apoptosis in the HepG2 cells, while AICAR, the AMPK activator, elicited marked cytotoxic effects.Together, these results suggested that fatsioside A‑induced apoptotic death requires AMPK activation in HepG2 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Liver Diseases, Hospital for Infectious Diseases of Jining, Jining, Shandong 272031, P.R. China.

ABSTRACT
Hepatocellular carcinoma (HCC) is one of the most malignant types of human primary tumor and has a poor prognosis, therefore, the development of novel therapeutic modalities is necessary. Fatsioside A is a novel baccharane‑type triterpenoid glycoside, which is extracted from the fruits of Fatsia japonica. Previous data has revealed that fatsioside A can exert growth inhibition, cell cycle arrest and induce apoptosis in human glioma cells. However, no detailed investigations have been performed to determine its action on human hepatocellular cells, and the exact mechanisms underlying the induction of apoptosis remain to be elucidated. The aim of the present study was to investigate the anticancer effect of fatsioside A in the HepG2 human HCC cell line, and to investigate the underlying mechanisms by focusing on the AMP‑activated protein kinase (AMPK) signaling cascade. The results of the present study demonstrated that fatsioside A induced apoptotic death of the human HepG2 HCC cells, which was associated with a marked activation of AMPK and increased expression of the downstream acetyl‑CoA carboxylase carboxylase. Inhibition of AMPK by RNA interference or by its inhibitor, compound C, suppressed fatsioside A‑induced caspase‑3 cleavage and apoptosis in the HepG2 cells, while AICAR, the AMPK activator, elicited marked cytotoxic effects. Together, these results suggested that fatsioside A‑induced apoptotic death requires AMPK activation in HepG2 cells.

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AMPK activator inhibits HepG2 cell survival. (A) HepG2 cells were either left untreated or treated with fatsioside A (80 µM) for 24 h, or the AMPK activator (AICAR; 1 mM) for 2 h. The levels of cleaved caspase-3 were assessed using western blot analysis. (B) Cell viability was examined using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Experiments were repeated three times and similar results were obtained. *P<0.05, vs. Ctrl. FA, fatsioside A; Ctrl, untreated control; AICAR, 5-aminoimidazole-4-carb oxyamide-1-β-D-ribofuranoside; ACC, acetyl-CoA carboxylase carboxylase.
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f4-mmr-12-04-5679: AMPK activator inhibits HepG2 cell survival. (A) HepG2 cells were either left untreated or treated with fatsioside A (80 µM) for 24 h, or the AMPK activator (AICAR; 1 mM) for 2 h. The levels of cleaved caspase-3 were assessed using western blot analysis. (B) Cell viability was examined using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Experiments were repeated three times and similar results were obtained. *P<0.05, vs. Ctrl. FA, fatsioside A; Ctrl, untreated control; AICAR, 5-aminoimidazole-4-carb oxyamide-1-β-D-ribofuranoside; ACC, acetyl-CoA carboxylase carboxylase.

Mentions: As shown in Fig. 3A, fatsioside A induced significant AMPK activation in the HepG2 cells, as the expression levels of p-AMPKα and the downstream ACC in HepG2 cells were significantly increased following fatsioside A treatment. Notably, AMPK inhibition by its inhibitor, compound C or by AMPKα-RNAi suppressed fatsioside A-induced loss of cell viability (Fig. 3B and C). Fatsioside A-induced apoptosis and cleavage of caspase-3 was also inhibited by AMPK inhibition (Fig. 3D and E). The AICAR AMPK activator also inhibited HepG2 cell survival (Fig. 4A and B). The above results indicated that activation of AMPK was involved in fatsioside A-induced cytotoxicity in HepG2 cells.


Fatsioside A‑induced apoptotic death of HepG2 cells requires activation of AMP‑activated protein kinase.

Zheng YS, Zhang JY, Zhang DH - Mol Med Rep (2015)

AMPK activator inhibits HepG2 cell survival. (A) HepG2 cells were either left untreated or treated with fatsioside A (80 µM) for 24 h, or the AMPK activator (AICAR; 1 mM) for 2 h. The levels of cleaved caspase-3 were assessed using western blot analysis. (B) Cell viability was examined using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Experiments were repeated three times and similar results were obtained. *P<0.05, vs. Ctrl. FA, fatsioside A; Ctrl, untreated control; AICAR, 5-aminoimidazole-4-carb oxyamide-1-β-D-ribofuranoside; ACC, acetyl-CoA carboxylase carboxylase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581823&req=5

f4-mmr-12-04-5679: AMPK activator inhibits HepG2 cell survival. (A) HepG2 cells were either left untreated or treated with fatsioside A (80 µM) for 24 h, or the AMPK activator (AICAR; 1 mM) for 2 h. The levels of cleaved caspase-3 were assessed using western blot analysis. (B) Cell viability was examined using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Experiments were repeated three times and similar results were obtained. *P<0.05, vs. Ctrl. FA, fatsioside A; Ctrl, untreated control; AICAR, 5-aminoimidazole-4-carb oxyamide-1-β-D-ribofuranoside; ACC, acetyl-CoA carboxylase carboxylase.
Mentions: As shown in Fig. 3A, fatsioside A induced significant AMPK activation in the HepG2 cells, as the expression levels of p-AMPKα and the downstream ACC in HepG2 cells were significantly increased following fatsioside A treatment. Notably, AMPK inhibition by its inhibitor, compound C or by AMPKα-RNAi suppressed fatsioside A-induced loss of cell viability (Fig. 3B and C). Fatsioside A-induced apoptosis and cleavage of caspase-3 was also inhibited by AMPK inhibition (Fig. 3D and E). The AICAR AMPK activator also inhibited HepG2 cell survival (Fig. 4A and B). The above results indicated that activation of AMPK was involved in fatsioside A-induced cytotoxicity in HepG2 cells.

Bottom Line: The results of the present study demonstrated that fatsioside A induced apoptotic death of the human HepG2 HCC cells, which was associated with a marked activation of AMPK and increased expression of the downstream acetyl‑CoA carboxylase carboxylase.Inhibition of AMPK by RNA interference or by its inhibitor, compound C, suppressed fatsioside A‑induced caspase‑3 cleavage and apoptosis in the HepG2 cells, while AICAR, the AMPK activator, elicited marked cytotoxic effects.Together, these results suggested that fatsioside A‑induced apoptotic death requires AMPK activation in HepG2 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Liver Diseases, Hospital for Infectious Diseases of Jining, Jining, Shandong 272031, P.R. China.

ABSTRACT
Hepatocellular carcinoma (HCC) is one of the most malignant types of human primary tumor and has a poor prognosis, therefore, the development of novel therapeutic modalities is necessary. Fatsioside A is a novel baccharane‑type triterpenoid glycoside, which is extracted from the fruits of Fatsia japonica. Previous data has revealed that fatsioside A can exert growth inhibition, cell cycle arrest and induce apoptosis in human glioma cells. However, no detailed investigations have been performed to determine its action on human hepatocellular cells, and the exact mechanisms underlying the induction of apoptosis remain to be elucidated. The aim of the present study was to investigate the anticancer effect of fatsioside A in the HepG2 human HCC cell line, and to investigate the underlying mechanisms by focusing on the AMP‑activated protein kinase (AMPK) signaling cascade. The results of the present study demonstrated that fatsioside A induced apoptotic death of the human HepG2 HCC cells, which was associated with a marked activation of AMPK and increased expression of the downstream acetyl‑CoA carboxylase carboxylase. Inhibition of AMPK by RNA interference or by its inhibitor, compound C, suppressed fatsioside A‑induced caspase‑3 cleavage and apoptosis in the HepG2 cells, while AICAR, the AMPK activator, elicited marked cytotoxic effects. Together, these results suggested that fatsioside A‑induced apoptotic death requires AMPK activation in HepG2 cells.

Show MeSH
Related in: MedlinePlus