Intrinsic anticarcinogenic effects of Piper sarmentosum ethanolic extract on a human hepatoma cell line
Authors: Sahidan Senafi
Wan Haifa Haryani Wan Omar
Muhd Fauzi Safian
Rohaya Megat Abdul Wahab
Shahrul Hisham Zainal Ariffin
Zaidah Zainal Ariffin
Journal: Cancer Cell International (2009)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Piper sarmentosum; anticarcinogenic activity; HepG2 cell line; ethanolic extract; IC50 value
Affiliations: School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Selangor Darul Ehsan, Malaysia. 

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 &lt; 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. 

Retracted: No

PubPeer comments:
 unlikely zero change between D and F ![file] Disclaimer: These potential image duplications have been detected automatically using ImageTwin http: dx.doi.org 10.1145 3019612.3019875 Although results have been curated, the automatic algorithm may have generated false positive results and cannot take into account all scientific explanations for apparent duplications like stripped and reprobed blots, time series, serial sections and reused controls. , 
 This is between treated and untreated cells from the same cell line (C vs D and E vs F) . Cellular picture of treated and untreated cell were choosen at similar cell count to compare the different between treated and untreated cell. , 
 #2 Thank you for your reply. To be sure I understand, are you saying the untreated cells showed no discernible difference between 48 and 72 hours (other than a change in magnification)? ,