Induction of apoptosis in human hepatocellular carcinoma cells by extracts of Lannea coromandelica (Houtt.) Merr. and Diospyros castanea (Craib) Fletcher.
BACKGROUND
Herbal plants are a preferred source of anticancer agents. This study aims to screen the anticancer activity of a crude extract of twigs of (a) Bombax anceps Pierre var. anceps (BA); (b) Catunaregam tomentosa (Blume ex DC.) Tirveng. (CT); (c) Erythrophleum succirubrum Gagnep. (ES); (d) Lannea coromandelica (Houtt.) Merr. (LC); and (e) leaves and (f) twigs of Diospyros castanea (Craib) Fletcher (DC).


METHODS
The 50 % ethanol-water extracts were prepared from each plant sample. In vitro anticancer effects of six extracts on the human hepatocellular carcinoma cell line (HepG2) in terms of cytotoxicity were investigated by neutral red assay, apoptosis induction by 4',6-diamidino-2-phenylindole (DAPI) staining, and DNA fragmentation by agarose gel electrophoresis. Normal Vero cells were tested for comparison and to determine cancer selectivity. Gas chromatography-mass spectrometry analysis was performed to identify the compounds in the extracts.


RESULTS
The six crude extracts had different cytotoxicities and were classified into three groups based on their IC50 value and selectivity index (SI). DC (twig) crude extract had both a high cytotoxicity and SI toward HepG2 cells comparable to melphalan (P = 0.023). The crude extracts of DC (leaves), LC (twig), and BA (twig) had moderate cytotoxicity and a lower SI. Although all crude plant extracts induced apoptosis in more than 50 % of the DAPI-positive apoptotic HepG2 cells, only DC (twig) and LC (twig) showed laddering in the DNA fragmentation assay. 2-Palmitoylglycerol was the major compound common to both. Pyrogallol and lupeol were the major compounds in DC (twig) crude extract. Hexadecanoic acid and octadecenoic acid were the major compounds in LC (twig) crude extract, which had high toxicity but low selectivity.


CONCLUSIONS
Ethanolic extracts from DC and LC twigs induced apoptosis in the HepG2 cell line. Pyrogallol and lupeol in DC (twig) might be responsible for the cytotoxicity toward the HepG2 cancer cells.
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002 Thailand.
Natthida
N
Weerapreeyakul
Graduate School, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002 Thailand.
Cholpajsorn
C
Junhom
Faculty of Associate Medical Sciences, Khon Kaen University, Khon Kaen, 40002 Thailand.
Sahapat
S
Barusrux
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002 Thailand.
Thaweesak
T
Thitimetharoch
Apoptosis; Cytotoxicity; Diospyros castanea; HepG2; Hepatocellular carcinoma; Lannea coromandelica