Ethanolic extract of Condurango (Marsdenia condurango) used in traditional systems of medicine including homeopathy against cancer can induce DNA damage and apoptosis in non small lung cancer cells, A549 and H522, in vitro
Sikdar Sourav, Mukherjee Avinaba, Boujedaini Naoual, Khuda Bukhsh Anisur Rahman,
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( Sikdar Sourav )
University of Kalyani Department of Zoology
( Mukherjee Avinaba )
University of Kalyani Department of Zoology
( Boujedaini Naoual )
Boiron Laboratory
( Khuda Bukhsh Anisur Rahman )
University of Kalyani Department of Zoology
KMID : 1140620130030010009
Abstract
In traditional systems of medicine including homeopathy, the Condurango extract (Con) is often used to cure stomach cancer mainly, without having any scientific validation of its anti-cancer ability. Con has therefore been tested against non-small-cell lung cancer cells (NSCLC) A549 and NCI-H522 (H522) known to contain the KRAS mutation, making them resistant to most chemotherapeutic agents. As cancer cells generally defy cytotoxicity developed by chemopreventive agents and escape cell death, any drug showing the capability of preferentially killing cancer cells through apoptosis is worth consideration for judicious application. A549 and H522 cells were exposed to 0.35 ¥ìg/¥ìl and 0.25 ¥ìg/¥ìl of Con, respectively, for 48 h and analysed based on various protocols associated with apoptosis and DNA damage, such as MTT assay to determine cell viability, LDH assay, DNA fragmentation assay, comet assay, and microscopical examinations of DNA binding fluorescence stains like DAPI, Hoechst 33258 and acridine orange/ethidium bromide to determine the extent of DNA damage made in drug-treated and untreated cells and the results compared. Changes in mitochondrial membrane potential and the generation of reactive oxygen species were also documented through standard techniques. Con killed almost 50% of the cancer cells but spared normal cells significantly. Fluorescence studies revealed increased DNA nick formation and depolarized membrane potentials after drug treatment in both cell types. Caspase-3 expression levels confirmed the apoptosis-inducing potential of Con in both the NSCLC lines. Thus, overall results suggest considerable anticancer potential of Con against NSCLC in vitro, validating its use against lung cancer by practitioners of traditional medicine including homeopathy.
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anticancer potential; Condurango extract; A549 and NCI-H522 non small-cell lung cancer cells; apoptosis; DNA damage; reactive oxygen species; mitochondrial membrane potential
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