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Thujone-Rich Fraction of Thuja occidentalis Demonstrates Major Anti-Cancer Potentials: Evidences from In Vitro Studies on A375 Cells

Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India.
Evidence-based Complementary and Alternative Medicine (Impact Factor: 1.88). 02/2011; 2011:568148. DOI: 10.1093/ecam/neq042
Source: PubMed

ABSTRACT CRUDE ETHANOLIC EXTRACT OF THUJA OCCIDENTALIS (FAM: Cupressaceae) is used as homeopathic mother tincture (TOΦ) to treat various ailments, particularly moles and tumors, and also used in various other systems of traditional medicine. Anti-proliferative and apoptosis-inducing properties of TOΦ and the thujone-rich fraction (TRF) separated from it have been evaluated for their possible anti-cancer potentials in the malignant melanoma cell line A375. On initial trial by S-diphenyltetrazolium bromide assay, both TOΦ and TRF showed maximum cytotoxic effect on A375 cell line while the other three principal fractions separated by chromatography had negligible or no such effect, because of which only TRF was further characterized and subjected to certain other assays for determining its precise anti-proliferative and apoptotic potentials. TRF was reported to have a molecular formula of C(10)H(16)O with a molecular weight of 152. Exposure of TRF of Thuja occidentalis to A375 cells in vitro showed more cytotoxic, anti-proliferative and apoptotic effects as compared with TOΦ, but had minimal growth inhibitory responses when exposed to normal cells (peripheral blood mononuclear cell). Furthermore, both TOΦ and TRF also caused a significant decrease in cell viability, induced inter-nucleosomal DNA fragmentation, mitochondrial transmembrane potential collapse, increase in ROS generation, and release of cytochrome c and caspase-3 activation, all of which are closely related to the induction of apoptosis in A375 cells. Thus, TRF showed and matched all the anti-cancer responses of TOΦ and could be the main bio-active fraction. The use of TOΦ in traditional medicines against tumors has, therefore, a scientific basis.

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    • "Essential oil of B. oriantalis has been widely used in steam bath. The thujone-rich fraction separated from crude ethanolic extract of Biota had been reported to possess anticancer potential (Biswas et al., 2011). Antibacterial, antifungal, analgesic, antinflammatory, anthelmintic and pharmacological effects of this plant has also been studied by some workers (Ezzat, 2001; Guleria et al., 2008; Bissa et al., 2008; Bhan et al., 2011). "
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    • "Essential oil of B. oriantalis has been widely used in steam bath. The thujone-rich fraction separated from crude ethanolic extract of Biota had been reported to possess anticancer potential (Biswas et al., 2011). Antibacterial, antifungal, analgesic, antinflammatory, anthelmintic and pharmacological effects of this plant has also been studied by some workers (Ezzat, 2001; Guleria et al., 2008; Bissa et al., 2008; Bhan et al., 2011). "
    Dataset: biota
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    • "Briefly, treated or untreated cells were stained with acridine orange (10 ␮g/ml) and ethidium bromide (10 ␮g/ml) and analyzed under fluorescence microscope with excitation at 488 nm and 550 nm. To detect nuclear damage or chromatin condensation, treated and untreated cells were washed twice with PBS and stained with 10 ␮g/ml of DAPI and observed under fluorescence microscope with excitation at 359 nm and emission at 461 nm using DAPI filter at 200× magnification (Biswas et al., 2011). "
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