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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 tamoxifen in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations (1.56–25 μg mL-1). The IC50 value for HepG2 is 3 μg mL-1 and 18.6 μg mL-1 for non-malignant Chang's liver cells. Each data point represent values from three independent experiments (n = 3).
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Figure 2: MTT assaying of tamoxifen in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations (1.56–25 μg mL-1). The IC50 value for HepG2 is 3 μg mL-1 and 18.6 μg mL-1 for non-malignant Chang's liver cells. Each data point represent values from three independent experiments (n = 3).

Mentions: Comparatively, tamoxifen, a drug with anti-oestrogenic activity, was used in this study as a positive control. Tamoxifen imposed an inhibitory effect in the HepG2 cell line with an IC50 value of 3 μg mL-1 and in the non-malignant Chang's liver cell line with a value of 18.6 μg mL-1(Figure 2). Therefore, tamoxifen induced cytotoxic activity in both carcinoma (HepG2) and non-carcinoma (non-malignant Chang's liver) cells. Both cells induced IC50 below 30 μg mL-1 and were thus considered to induce cytotoxic activity to the treated cells, as recommended by National Cancer Institute (NCI) [19]. NCI recommended that any extract generates IC50 below than 30 μg mL-1 is considered possess cytotoxic activity. As a result, MTT assay analysis showed that the ethanolic extract of P. sarmentosum induced cytotoxic activity in HepG2 cells, but not in the non-malignant Chang's liver cells. In contrast, an anticarcinogenic drug (tamoxifen) induced cytotoxic activity in hepatocellular carcinoma, HepG2 and non-malignant Chang's liver cell lines.


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 tamoxifen in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations (1.56–25 μg mL-1). The IC50 value for HepG2 is 3 μg mL-1 and 18.6 μg mL-1 for non-malignant Chang's liver cells. Each data point represent 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 2: MTT assaying of tamoxifen in HepG2 and non-malignant Chang's liver cells. Both cells were treated at various concentrations (1.56–25 μg mL-1). The IC50 value for HepG2 is 3 μg mL-1 and 18.6 μg mL-1 for non-malignant Chang's liver cells. Each data point represent values from three independent experiments (n = 3).
Mentions: Comparatively, tamoxifen, a drug with anti-oestrogenic activity, was used in this study as a positive control. Tamoxifen imposed an inhibitory effect in the HepG2 cell line with an IC50 value of 3 μg mL-1 and in the non-malignant Chang's liver cell line with a value of 18.6 μg mL-1(Figure 2). Therefore, tamoxifen induced cytotoxic activity in both carcinoma (HepG2) and non-carcinoma (non-malignant Chang's liver) cells. Both cells induced IC50 below 30 μg mL-1 and were thus considered to induce cytotoxic activity to the treated cells, as recommended by National Cancer Institute (NCI) [19]. NCI recommended that any extract generates IC50 below than 30 μg mL-1 is considered possess cytotoxic activity. As a result, MTT assay analysis showed that the ethanolic extract of P. sarmentosum induced cytotoxic activity in HepG2 cells, but not in the non-malignant Chang's liver cells. In contrast, an anticarcinogenic drug (tamoxifen) induced cytotoxic activity in hepatocellular carcinoma, HepG2 and non-malignant Chang's liver cell lines.

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