Investigation of Cytotoxic Activity on Human Cancer Cell Lines of Arborinine and Furanoacridones Isolated from Ruta graveolens

Abstract

The cytotoxic effects of a series of furanoacridones isolated from Ruta graveolens L. (Rutaceae) and of two further acridone alkaloids (arborinine and evoxanthine) were investigated by means of the MTT assay, using the human cell lines HeLa, MCF7 and A431. Arborinine proved best in inhibiting the proliferation of all three cell lines. The cytotoxic potency of the furacridone alkaloids was a function of their lipid solubility, which was determined by means of PAMPA. The capacity of the most effective furanoacridones to induce apoptosis was demonstrated by flow cytometric cell cycle analysis and by staining with ethidium bromide and acridine orange. This finding was reinforced by determining the apoptosis-regulating factors Bcl-2 and Bax, which were revealed by means of RT-PCR to change dose-dependently. The data presented here indicate that naturally occurring furanoacridones can be regarded as excellent starting structures for the potential development of new anticancer agents.

Abbreviations

AO:acridine orange

cDNA:copy deoxyribonucleic acid

DMSO:dimethyl sulfoxide

dNTP:deoxyribonucleotide triphosphate

EB:ethidium bromide

hGAPDH:human glyceraldehyde-3-phosphate dehydrogenase

MMLV-RT:moloney murine leukemia virus-reverse transcriptase

mRNA:messenger ribonucleic acid

MTT:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PAMPA:parallel artificial membrane permeability assays

PBS:phosphate buffered saline

RNA:ribonucleic acid

RT-PCR:reverse transcription-polymerase chain reaction

Key words

Ruta graveolens L. - Rutaceae - acridone alkaloids - cytotoxicity - apoptosis

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