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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

Morphological observation with May-Grunwald-Giemsa's staining at actual magnification 100×. HepG2 cells were treated for 24 (A), 48 (C) and 72 (E) hours with 12.5 μg mL-1 of P. sarmentosum ethanolic extract while untreated HepG2 cells were grow in complete medium for 24 (B), 48 (D) and 72 (F) hours. The white arrows indicated apoptotic bodies. The figures shown are representative of three independent experiments (n = 3).
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Figure 4: Morphological observation with May-Grunwald-Giemsa's staining at actual magnification 100×. HepG2 cells were treated for 24 (A), 48 (C) and 72 (E) hours with 12.5 μg mL-1 of P. sarmentosum ethanolic extract while untreated HepG2 cells were grow in complete medium for 24 (B), 48 (D) and 72 (F) hours. The white arrows indicated apoptotic bodies. The figures shown are representative of three independent experiments (n = 3).

Mentions: Ethanolic extracts from P. sarmentosum can induce apoptosis in HepG2 cells, as proven using May-Grunwald-Giemsa's staining (Figure 4). The apoptotic morphological pictures clearly show the appearance of apoptotic bodies (indicated as white arrow) when using an inverted microscope at 100× actual magnification. Marked morphological changes of the apoptotic cells are represented by apoptotic bodies (indicated as white arrow), which are easily determined by May-Grunwald-Giemsa's staining (Figure 4A, C and 4E). These apoptotic cells can be seen when the cells are exposed to 12.5 μg mL-1 of the ethanolic extract for 24, 48 and 72 hours.


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)

Morphological observation with May-Grunwald-Giemsa's staining at actual magnification 100×. HepG2 cells were treated for 24 (A), 48 (C) and 72 (E) hours with 12.5 μg mL-1 of P. sarmentosum ethanolic extract while untreated HepG2 cells were grow in complete medium for 24 (B), 48 (D) and 72 (F) hours. The white arrows indicated apoptotic bodies. The figures shown are representative of three independent experiments (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Morphological observation with May-Grunwald-Giemsa's staining at actual magnification 100×. HepG2 cells were treated for 24 (A), 48 (C) and 72 (E) hours with 12.5 μg mL-1 of P. sarmentosum ethanolic extract while untreated HepG2 cells were grow in complete medium for 24 (B), 48 (D) and 72 (F) hours. The white arrows indicated apoptotic bodies. The figures shown are representative of three independent experiments (n = 3).
Mentions: Ethanolic extracts from P. sarmentosum can induce apoptosis in HepG2 cells, as proven using May-Grunwald-Giemsa's staining (Figure 4). The apoptotic morphological pictures clearly show the appearance of apoptotic bodies (indicated as white arrow) when using an inverted microscope at 100× actual magnification. Marked morphological changes of the apoptotic cells are represented by apoptotic bodies (indicated as white arrow), which are easily determined by May-Grunwald-Giemsa's staining (Figure 4A, C and 4E). These apoptotic cells can be seen when the cells are exposed to 12.5 μg mL-1 of the ethanolic extract for 24, 48 and 72 hours.

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