Accumulation of betulinic, oleanolic, and ursolic acids in In vitro cell cultures of Lantana camara L. and their significant cytotoxic effects on HeLa cell lines

Abstract

This study reports the accumulation of three pentacyclic triterpenoids, betulinic acid (BA), oleanolic acid (OA), and ursolic acid (UA), in cell cultures of Lantana camara using leaf disc explants. Thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and mass spectroscopy (MS) of organic extract followed by comparison with the standards revealed the presence of the above three triterpenoids. By following the extraction procedure mentioned here the recovery of all these three compounds were > 95%. The bioactivity of the cell-derived extract was demonstrated using cancerous HeLa cell lines. Specifically, the effect of this extract on HeLa cells was noticed from 36 h (at 100 μg/mL) to 72 h (at 25 μg/mL) by employing 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay. The results show that an increase in the concentration or duration of the extract treatment was effective in killing cancerous cells. DNA laddering assay confirmed that cell death was due to apoptosis. Further, BA was identified for the first time from an in vitro source. Moreover, this was the first report describing this kind of studies conducted using Lantana plants.