Apoptosis-Induced Cell Death due to Oleanolic Acid in HaCaT Keratinocyte Cells -a Proof-of-Principle Approach for Chemopreventive Drug Development

Abstract

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells -a cell-based epithelial model system for toxicity/ethnopharmacology-based studies -was conducted. Radical scavenging activity (DPPH•) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 µM) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

Antitumor Constituents from Anthriscus Sylvestris (L.) Hoffm

“…Syzygium aromaticum L showed inhibition activity of some cancer cell lines like HeLa (Dwivedi et al, 2011). In addition, some compounds like oleanolic acid and ursolic acid from food and medicinal herbs had relatively low toxicity and antitumor activities (Gayathri et al, 2009;George et al, 2012). In this study, the bioassayguided chemical investigation of the roots of A. sylvestris resulted in the isolation of nine known compounds, and their antitumor activities were tested.…”

Triterpenes as Potentially Cytotoxic Compounds

“…Activity of the triterpenes was evaluated with the use of MTT and SRB tests, showing that easy modifications may transform the compounds into agents manifesting some promising anti-cancer activity [75]. The oleanolic acid is also capable of inducing apoptosis in line HaCaT (immortalized keratinocyte cell line) at relatively low cytotoxicity, which points to potential anti-cancer properties of this compound [76]. As in case of other pentacyclic triterpenes, synthetized and looked for are derivatives of the oleanolic acid, supposed to show stronger cytotoxicity and selectivity as well as better capabilities of inducing apoptosis.…”

Research Article Anticancer and antimicrobial activities of diosmin

“…Dosedependent and time-dependent increase in the DNA fragmentation of diosmin-treated MDA-MB-231 cells, determined by the increase in absorbance values, should indicate that diosmin causes apoptosis by inducing DNA damage in this aggressive cell type. Apoptosis is generally characterized by morphological changes in apoptotic cells, such as volume reduction, condensation of the nucleus, swelling of the cytoplasm, loss of membrane integrity followed by plasma membrane blebbing, and finally the formation of apoptotic bodies (Choi et al, 2012;George et al, 2012). Morphological change in MDA-MB-231 cells was observed under an inverted microscope after exposure to diosmin at IC 50 .…”