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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 studies by inverted microscope at actual magnification 100×. HepG2 and non-malignant Chang's liver cell line were treated without (A, C) and with 12.5 μg mL-1 of P. sarmentosum ethanolic extract (B, D) for 72 hours. Both types of treatment (C and D) produced similar cellular morphology and antiproliferative effect. However, in HepG2 cells, the confluency appeared to be reduced from 90% in untreated cells to 10% in treated cells. Similar cellular morphology was observed in three independent experiments (n = 3).
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Figure 3: Morphological studies by inverted microscope at actual magnification 100×. HepG2 and non-malignant Chang's liver cell line were treated without (A, C) and with 12.5 μg mL-1 of P. sarmentosum ethanolic extract (B, D) for 72 hours. Both types of treatment (C and D) produced similar cellular morphology and antiproliferative effect. However, in HepG2 cells, the confluency appeared to be reduced from 90% in untreated cells to 10% in treated cells. Similar cellular morphology was observed in three independent experiments (n = 3).

Mentions: Light microscopic observation of the P. sarmentosum ethanolic extract-treated HepG2 cell line after 72 hours of exposure showed typical morphological features of apoptosis. The characterisation of morphological changes observed were reduction in cell volume, cell shrinkage, reduction in chromatin condensation and formation of cytoplasmic blebs [20]. Figure 3B shows that the HepG2 cells treated with ethanolic extract at 12.5 μg mL-1were changed into round shapes as compared to untreated HepG2 cells (Figure 3A). The untreated cells (HepG2) also showed a high confluency of monolayer cells (Figure 3A) compared to ethanolic extract-treated cells, which showed a reduction in cell volume and cell shrinkage (Figure 3B). Figure 3C shows that the morphology of the untreated non-malignant Chang's liver cell line is a confluent monolayer. The non-malignant Chang's liver cell line was then treated with 12.5 μg mL-1 of P. sarmentosum ethanolic extract. After 72 hours of incubation, the morphology of the treated non-malignant Chang's liver cell line (Figure 3D) showed similar morphology to that untreated non-malignant Chang's liver cell line (Figure 3C).


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 studies by inverted microscope at actual magnification 100×. HepG2 and non-malignant Chang's liver cell line were treated without (A, C) and with 12.5 μg mL-1 of P. sarmentosum ethanolic extract (B, D) for 72 hours. Both types of treatment (C and D) produced similar cellular morphology and antiproliferative effect. However, in HepG2 cells, the confluency appeared to be reduced from 90% in untreated cells to 10% in treated cells. Similar cellular morphology was observed in three independent experiments (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Morphological studies by inverted microscope at actual magnification 100×. HepG2 and non-malignant Chang's liver cell line were treated without (A, C) and with 12.5 μg mL-1 of P. sarmentosum ethanolic extract (B, D) for 72 hours. Both types of treatment (C and D) produced similar cellular morphology and antiproliferative effect. However, in HepG2 cells, the confluency appeared to be reduced from 90% in untreated cells to 10% in treated cells. Similar cellular morphology was observed in three independent experiments (n = 3).
Mentions: Light microscopic observation of the P. sarmentosum ethanolic extract-treated HepG2 cell line after 72 hours of exposure showed typical morphological features of apoptosis. The characterisation of morphological changes observed were reduction in cell volume, cell shrinkage, reduction in chromatin condensation and formation of cytoplasmic blebs [20]. Figure 3B shows that the HepG2 cells treated with ethanolic extract at 12.5 μg mL-1were changed into round shapes as compared to untreated HepG2 cells (Figure 3A). The untreated cells (HepG2) also showed a high confluency of monolayer cells (Figure 3A) compared to ethanolic extract-treated cells, which showed a reduction in cell volume and cell shrinkage (Figure 3B). Figure 3C shows that the morphology of the untreated non-malignant Chang's liver cell line is a confluent monolayer. The non-malignant Chang's liver cell line was then treated with 12.5 μg mL-1 of P. sarmentosum ethanolic extract. After 72 hours of incubation, the morphology of the treated non-malignant Chang's liver cell line (Figure 3D) showed similar morphology to that untreated non-malignant Chang's liver cell line (Figure 3C).

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