The aim of the current study was to evaluate the anticancer potential of tamarind bark and its bioactive fraction on HeLa and PA-1 cell lines. Tamarind bark extract was prepared with dichloromethane as the solvent using Soxhlet apparatus and treated on HeLa and PA-1 cell lines to determine the anticancer potential by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Further, bioassay guided fractionation of the extract was carried out using thin layer chromatography. In vitro assays like fluorescence microscopy, DNA fragmentation analysis, caspase-9 activity assay and flow cytometry analyses were carried out to determine the mechanism of anticancer activity of the selected bioactive fraction. The fraction was further characterized by gas chromatography-mass spectrometry. Tamarind bark dichloromethane extract was found to be cytotoxic to HeLa and PA-1 cells. Bioassay guided fractionation indicated that the fourth fraction of the extract possessed the bioactive component. Further, in vitro assays demonstrated that the bioactive fraction induced caspase-9-mediated apoptosis in the cells and was able to reduce the total cell count as evidenced by flow cytometry analyses. The gas chromatography-mass spectrometry analysis of the bioactive fraction indicated the presence of cantharidin, an anticancer compound earlier reported from the blister beetles. It can be concluded that the presence of cantharidin might be responsible for inhibition of proliferation and induction of apoptosis in the cancer cells. Key words: Apoptosis, anticancer, cantharidin, caspase, tamarind bark extractCancer is one of the most dreaded diseases as mortality associated is very high despite the availability of modern treatment modalities [1,2] . Hence, research towards novel, more effective and tumor-specific treatment has gained momentum during the past decades. In this pursuit, several medicinal and dietary plants have provided many anticancer compounds till date [3,4] . Tamarindus indica L. commonly known as tamarind is one of the most important dietary plants in which all parts have some nutritional or medicinal value. It has a history that dates back to ancient tribal people who used to prepare decoctions of the parts and use it for medication. It was reported that tamarind bark and leaves were used to treat wounds in West and East Africa [5] . Fruit and leaf have laxative property, paste of dried seeds were used to set fractured bones, and decoction of the bark and leaves were used to treat jaundice [6][7][8] . The bark of tamarind tree was reported as to have antiinflammatory, analgesic and wound healing properties [9,10] . In West Africa, the macerated fresh bark of the young twigs was used both as a purgative and to relieve abdominal pain [11] .Cantharidin is a colorless, odourless terpene secreted by many species of blister beetles. Cantharidin, isolated from the Chinese blister beetles, Mylabris phalerata or M. cichorii, displays antitumor activity and induces apoptosis in many types of tumor cells. It can be poisono...
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