Cytotoxic and apoptotic effects of six herbal plants against the human hepatocarcinoma (HepG2) cell line

<p>Abstract</p> <p>Background</p> <p>Six plants from Thailand were evaluated for their cytotoxicity and apoptosis induction in human hepatocarcinoma (HepG2) as compared to normal African green monkey kidney epithelial cell lines.</p> <p>Methods</p> <p>Ethanol-water crude extracts of the six plants were tested with neutral red assay for their cytotoxicity after 24 hours of exposure to the cells. Apoptotic induction was tested in the HepG2 cells with diamidino-2-phenylindole staining. DNA fragmentation, indicative of apoptosis, was analyzed with agarose gel electrophoresis. Alkylation, indicative of DNA damage, was also evaluated <it>in vitro </it>by 4-(4'-nitrobenzyl) pyridine assay.</p> <p>Results</p> <p>The extract of <it>Pinus kesiya </it>showed the highest selectivity (selectivity index = 9.6) and potent cytotoxicity in the HepG2 cell line, with an IC₅₀value of 52.0 ± 5.8 μg/ml (mean ± standard deviation). Extract of <it>Catimbium speciosum </it>exerted cytotoxicity with an IC₅₀value of 55.7 ± 8.1 μg/ml. Crude extracts from <it>Glochidion daltonii</it>, <it>Cladogynos orientalis</it>, <it>Acorus tatarinowii </it>and <it>Amomum villosum </it>exhibited cytotoxicity with IC₅₀values ranging 100-500 μg/ml. All crude extracts showed different alkylating abilities <it>in vitro</it>. Extracts of <it>P. kesiya, C. speciosum </it>and <it>C. orientalis </it>caused nuclei morphological changes and DNA laddering.</p> <p>Conclusion</p> <p>The extracts of <it>C. speciosum</it>, <it>C. orientalis </it>and <it>P. kesiya </it>induced apoptosis. Among the three plants, <it>P. kesiya </it>possessed the most robust anticancer activity, with specific selectivity against HepG2 cells.</p

Evaluation of the anticancer potential of six herbs against a hepatoma cell line

Abstract Background Six herbs in the Plant Genetics Conservation Project that have been used as complementary medicines were chosen on the basis of their medicinal value, namely Terminalia mucronata, Diospyros winitii, Bridelia insulana, Artabotrys harmandii, Terminallia triptera, and Croton oblongifolius. This study aims to evaluate the potential anticancer activity of 50% ethanol-water extracts of these six herbs. Methods Fifty percent ethanol-water crude extracts of the six herbs were prepared. The cytotoxicity of the herbal extracts relative to that of melphalan was evaluated using a hepatoma cell line (HepG2), and examined by neutral red assays and apoptosis induction by gel electrophoresis and flow cytometry after 24 h. Results A significant difference was found between the cytotoxicity of the 50% ethanol-water crude extracts and melphalan (P = 0.000). The 50% ethanol-water crude extracts of all six herbs exhibited cytotoxicity against HepG2 cells, with IC50 values ranging from 100 to 500 μg/mL. The extract of T. triptera showed the highest cytotoxicity with an IC50 of 148.7 ± 12.3 μg/mL, while melphalan had an IC50 of 39.79 ± 7.62 μg/mL. The 50% ethanol-water crude extracts of D. winitii and T. triptera, but not A. harmandii, produced a DNA ladder. The 50% ethanol-water crude extracts of D. winitii, T. triptera, and A. harmandii induced apoptosis detected by flow cytometry. Conclusion The 50% ethanol-water crude extracts of D. winitii, T. triptera, and A. harmandii showed anticancer activity in vitro.</p

FTIR Microspectroscopy for the Assessment of Mycoplasmas in HepG2 Cell Culture

To assess the presence and absence of mycoplasma contamination in cell culture, Fourier transform infrared (FTIR) microspectroscopy, coupled with multivariate analysis, was deployed to determine the biomolecular changes in hepatocellular carcinoma cells, HepG2, before and after mycoplasma contamination. The contaminated HepG2 cells were treated with antibiotic BM-Cyclin to decontaminate the mycoplasma, and polymerase chain reaction (PCR) was then performed to confirm the presence or the absence of mycoplasma contamination. The contaminated and decontaminated HepG2 cells were analyzed by FTIR microspectroscopy with principal component analysis (PCA) and peak integral area analysis. The results showed that the FTIR spectra of contaminated HepG2 cells demonstrated the alteration in the IR spectra corresponding to the lipid, protein, and nucleic acid regions. PCA analysis distinguished the spectral differences between the groups of mycoplasma-contaminated and -decontaminated cells. The PCA loading plots suggest that lipid and protein are the main contributed molecules for the difference between these two cell groups. Peak integral area analysis illustrated the increase of lipid and nucleic acid and the decrease of protein contents in the contaminated HepG2 cells. FTIR microspectroscopy is, therefore, proven to be a potential tool for assessing mycoplasma removal by monitoring biomolecular alterations in cell culture