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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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Fatsioside A inhibits the survival and proliferation of HepG2 cells. (A) HepG2 cells were either left untreated or were treated with different concentrations of fatsioside A, and the cells were further cultured in Dulbecco's modified Eagle's medium for 24 h. Cell viability was assessed using a 3-(4, 5-dimetryl-thiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. (B) Percentages of trypan blue-positive cells were recorded. (C) HepG2 cell proliferation was analyzed using a BrdU incorporation assay. (D) Apoptotic death of the HepG2 cells was determined using a TUNEL assay. Scale bar=100 µm. Experiments were repeated three times and similar results were obtained. Data are expressed as the mean ± standard deviation. *P<0.05, vs. Ctrl group. FA, fatsioside A; Ctrl, untreated control; OD, optical density.
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f1-mmr-12-04-5679: Fatsioside A inhibits the survival and proliferation of HepG2 cells. (A) HepG2 cells were either left untreated or were treated with different concentrations of fatsioside A, and the cells were further cultured in Dulbecco's modified Eagle's medium for 24 h. Cell viability was assessed using a 3-(4, 5-dimetryl-thiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. (B) Percentages of trypan blue-positive cells were recorded. (C) HepG2 cell proliferation was analyzed using a BrdU incorporation assay. (D) Apoptotic death of the HepG2 cells was determined using a TUNEL assay. Scale bar=100 µm. Experiments were repeated three times and similar results were obtained. Data are expressed as the mean ± standard deviation. *P<0.05, vs. Ctrl group. FA, fatsioside A; Ctrl, untreated control; OD, optical density.

Mentions: The present study examined the effect of fatsioside A on HepG2 cell survival and proliferation. Cell viability was evaluated using an MTT assay. The results, as shown in Fig. 1A, clearly indicated that fatsioside A at concentrations of 40 and 80 µM markedly inhibited HepG2 cell survival. In addition, the number of trypan blue-positive, 'dead', cells increased sharply following fatsioside A treatment at 40 and 80 µM (Fig. 1B). The effect of fatsioside A on HepG2 cell proliferation was also examined. Using a BrdU incorporation assay, the present study demonstrated that fatsioside A suppressed HepG2 cell proliferation (Fig. 1C). Furthermore, a TUNEL assay revealed that, compared with the control group, the number of tunnel-positive 'dead' cells increased markedly following fatsioside A treatment at concentrations of 40 and 80 µM (Fig. 1D). Taken together, these results suggested that fatsioside A significantly inhibited the survival and proliferation of the 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)

Fatsioside A inhibits the survival and proliferation of HepG2 cells. (A) HepG2 cells were either left untreated or were treated with different concentrations of fatsioside A, and the cells were further cultured in Dulbecco's modified Eagle's medium for 24 h. Cell viability was assessed using a 3-(4, 5-dimetryl-thiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. (B) Percentages of trypan blue-positive cells were recorded. (C) HepG2 cell proliferation was analyzed using a BrdU incorporation assay. (D) Apoptotic death of the HepG2 cells was determined using a TUNEL assay. Scale bar=100 µm. Experiments were repeated three times and similar results were obtained. Data are expressed as the mean ± standard deviation. *P<0.05, vs. Ctrl group. FA, fatsioside A; Ctrl, untreated control; OD, optical density.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1-mmr-12-04-5679: Fatsioside A inhibits the survival and proliferation of HepG2 cells. (A) HepG2 cells were either left untreated or were treated with different concentrations of fatsioside A, and the cells were further cultured in Dulbecco's modified Eagle's medium for 24 h. Cell viability was assessed using a 3-(4, 5-dimetryl-thiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. (B) Percentages of trypan blue-positive cells were recorded. (C) HepG2 cell proliferation was analyzed using a BrdU incorporation assay. (D) Apoptotic death of the HepG2 cells was determined using a TUNEL assay. Scale bar=100 µm. Experiments were repeated three times and similar results were obtained. Data are expressed as the mean ± standard deviation. *P<0.05, vs. Ctrl group. FA, fatsioside A; Ctrl, untreated control; OD, optical density.
Mentions: The present study examined the effect of fatsioside A on HepG2 cell survival and proliferation. Cell viability was evaluated using an MTT assay. The results, as shown in Fig. 1A, clearly indicated that fatsioside A at concentrations of 40 and 80 µM markedly inhibited HepG2 cell survival. In addition, the number of trypan blue-positive, 'dead', cells increased sharply following fatsioside A treatment at 40 and 80 µM (Fig. 1B). The effect of fatsioside A on HepG2 cell proliferation was also examined. Using a BrdU incorporation assay, the present study demonstrated that fatsioside A suppressed HepG2 cell proliferation (Fig. 1C). Furthermore, a TUNEL assay revealed that, compared with the control group, the number of tunnel-positive 'dead' cells increased markedly following fatsioside A treatment at concentrations of 40 and 80 µM (Fig. 1D). Taken together, these results suggested that fatsioside A significantly inhibited the survival and proliferation of the 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