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Intrinsic anticarcinogenic effects of Piper sarmentosum ethanolic extract on a human hepatoma cell line.

Zainal Ariffin SH, Wan Omar WH, Zainal Ariffin Z, Safian MF, Senafi S, Megat Abdul Wahab R - Cancer Cell Int. (2009)

Bottom Line: The percentage of apoptotic cells in the overall population (apoptotic index) showed a continuously significant increase (p < 0.05) in 12.5 mug mL-1 ethanolic extract-treated cells at 24, 48 and 72 hours compared to controls (untreated cells).These results showed a typical intrinsic apoptotic characterisation, which included fragmentation of nuclear DNA in ethanolic extract-treated HepG2 cells.Therefore, our results suggest that the ethanolic extract from P. sarmentosum induced anticarcinogenic activity through an intrinsic apoptosis pathway in HepG2 cells in vitro.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Selangor Darul Ehsan, Malaysia. shahroy8@gmail.com

ABSTRACT

Background: Piper sarmentosum, locally known as kaduk is belonging to the family of Piperaceae. It is our interest to evaluate their effect on human hepatoma cell line (HepG2) for the potential of anticarcinogenic activity.

Results: The anticarcinogenic activity of an ethanolic extract from Piper sarmentosum in HepG2 and non-malignant Chang's liver cell lines has been previously determined using (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) (MTT) assays, where the IC50 value was used as a parameter for cytotoxicity. The ethanolic extract that showed anticarcinogenic properties in HepG2 cells had an IC50 of 12.5 mug mL-1, while IC50 values in the non-malignant Chang's liver cell line were greater than 30 mug mL-1. Apoptotic morphological changes in HepG2 cells were observed using an inverted microscope and showed chromatin condensation, cell shrinkage and apoptotic bodies following May-Grunwald-Giemsa's staining. The percentage of apoptotic cells in the overall population (apoptotic index) showed a continuously significant increase (p < 0.05) in 12.5 mug mL-1 ethanolic extract-treated cells at 24, 48 and 72 hours compared to controls (untreated cells). Following acridine orange and ethidium bromide staining, treatment with 10, 12 and 14 mug mL-1 of ethanolic extracts caused typical apoptotic morphological changes in HepG2 cells. Molecular analysis of DNA fragmentation was used to examine intrinsic apoptosis induced by the ethanolic extracts. These results showed a typical intrinsic apoptotic characterisation, which included fragmentation of nuclear DNA in ethanolic extract-treated HepG2 cells. However, the non-malignant Chang's liver cell line produced no DNA fragmentation. In addition, the DNA genome was similarly intact for both the untreated non-malignant Chang's liver and HepG2 cell lines.

Conclusion: Therefore, our results suggest that the ethanolic extract from P. sarmentosum induced anticarcinogenic activity through an intrinsic apoptosis pathway in HepG2 cells in vitro.

No MeSH data available.


Related in: MedlinePlus

MTT assaying of the P. sarmentosum ethanolic extract in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations, i.e., 1.56–200 μg mL-1. The IC50 value for HepG2 was 12.5 μg mL-1, while the IC50 value for non-malignant Chang's liver cells was > 30 μg mL-1. Each data point represents values from three independent experiments (n = 3).
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Figure 1: MTT assaying of the P. sarmentosum ethanolic extract in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations, i.e., 1.56–200 μg mL-1. The IC50 value for HepG2 was 12.5 μg mL-1, while the IC50 value for non-malignant Chang's liver cells was > 30 μg mL-1. Each data point represents values from three independent experiments (n = 3).

Mentions: The criterion for cytotoxicity for the crude extracts, as established by the National Cancer Institute (NCI), is an IC50 value lower than 30 μg mL-1 [19]. Figure 1 showed that the P. sarmentosum ethanolic extract was able to exert antiproliferative effects in the HepG2 cell line tested in dose-dependent manner. The IC50 value of the ethanolic extract for HepG2 cells viability was 12.5 μg mL-1 after exposure for 72 hours (Figure 1). Our results show that the normal counterpart cells (non-malignant Chang's liver) treated with 200 μg mL-1 of ethanolic extract still retained > 50% viable cells, i.e., 55.6% viability. On the other hand, IC50 values for the ethanolic extract in non-malignant Chang's liver cells were more than 30 μg mL-1 (Figure 1). Therefore, P. sarmentosum ethanolic extract predetermination by MTT assay induced cytotoxicity activity in the hepatoma cell line (HepG2), but not in the non-malignant cell line (Chang's liver).


Intrinsic anticarcinogenic effects of Piper sarmentosum ethanolic extract on a human hepatoma cell line.

Zainal Ariffin SH, Wan Omar WH, Zainal Ariffin Z, Safian MF, Senafi S, Megat Abdul Wahab R - Cancer Cell Int. (2009)

MTT assaying of the P. sarmentosum ethanolic extract in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations, i.e., 1.56–200 μg mL-1. The IC50 value for HepG2 was 12.5 μg mL-1, while the IC50 value for non-malignant Chang's liver cells was > 30 μg mL-1. Each data point represents values from three independent experiments (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2667431&req=5

Figure 1: MTT assaying of the P. sarmentosum ethanolic extract in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations, i.e., 1.56–200 μg mL-1. The IC50 value for HepG2 was 12.5 μg mL-1, while the IC50 value for non-malignant Chang's liver cells was > 30 μg mL-1. Each data point represents values from three independent experiments (n = 3).
Mentions: The criterion for cytotoxicity for the crude extracts, as established by the National Cancer Institute (NCI), is an IC50 value lower than 30 μg mL-1 [19]. Figure 1 showed that the P. sarmentosum ethanolic extract was able to exert antiproliferative effects in the HepG2 cell line tested in dose-dependent manner. The IC50 value of the ethanolic extract for HepG2 cells viability was 12.5 μg mL-1 after exposure for 72 hours (Figure 1). Our results show that the normal counterpart cells (non-malignant Chang's liver) treated with 200 μg mL-1 of ethanolic extract still retained > 50% viable cells, i.e., 55.6% viability. On the other hand, IC50 values for the ethanolic extract in non-malignant Chang's liver cells were more than 30 μg mL-1 (Figure 1). Therefore, P. sarmentosum ethanolic extract predetermination by MTT assay induced cytotoxicity activity in the hepatoma cell line (HepG2), but not in the non-malignant cell line (Chang's liver).

Bottom Line: The percentage of apoptotic cells in the overall population (apoptotic index) showed a continuously significant increase (p < 0.05) in 12.5 mug mL-1 ethanolic extract-treated cells at 24, 48 and 72 hours compared to controls (untreated cells).These results showed a typical intrinsic apoptotic characterisation, which included fragmentation of nuclear DNA in ethanolic extract-treated HepG2 cells.Therefore, our results suggest that the ethanolic extract from P. sarmentosum induced anticarcinogenic activity through an intrinsic apoptosis pathway in HepG2 cells in vitro.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Selangor Darul Ehsan, Malaysia. shahroy8@gmail.com

ABSTRACT

Background: Piper sarmentosum, locally known as kaduk is belonging to the family of Piperaceae. It is our interest to evaluate their effect on human hepatoma cell line (HepG2) for the potential of anticarcinogenic activity.

Results: The anticarcinogenic activity of an ethanolic extract from Piper sarmentosum in HepG2 and non-malignant Chang's liver cell lines has been previously determined using (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) (MTT) assays, where the IC50 value was used as a parameter for cytotoxicity. The ethanolic extract that showed anticarcinogenic properties in HepG2 cells had an IC50 of 12.5 mug mL-1, while IC50 values in the non-malignant Chang's liver cell line were greater than 30 mug mL-1. Apoptotic morphological changes in HepG2 cells were observed using an inverted microscope and showed chromatin condensation, cell shrinkage and apoptotic bodies following May-Grunwald-Giemsa's staining. The percentage of apoptotic cells in the overall population (apoptotic index) showed a continuously significant increase (p < 0.05) in 12.5 mug mL-1 ethanolic extract-treated cells at 24, 48 and 72 hours compared to controls (untreated cells). Following acridine orange and ethidium bromide staining, treatment with 10, 12 and 14 mug mL-1 of ethanolic extracts caused typical apoptotic morphological changes in HepG2 cells. Molecular analysis of DNA fragmentation was used to examine intrinsic apoptosis induced by the ethanolic extracts. These results showed a typical intrinsic apoptotic characterisation, which included fragmentation of nuclear DNA in ethanolic extract-treated HepG2 cells. However, the non-malignant Chang's liver cell line produced no DNA fragmentation. In addition, the DNA genome was similarly intact for both the untreated non-malignant Chang's liver and HepG2 cell lines.

Conclusion: Therefore, our results suggest that the ethanolic extract from P. sarmentosum induced anticarcinogenic activity through an intrinsic apoptosis pathway in HepG2 cells in vitro.

No MeSH data available.


Related in: MedlinePlus