In vitro anti-breast cancer activity of ethanolic extract of Wrightia tomentosa: Role of pro-apoptotic effects of oleanolic acid and urosolic acid |
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Authors: | Bandana Chakravarti Ranjani Maurya Jawed Akhtar Siddiqui Hemant Kumar Bid SM Rajendran Prem P Yadav Rituraj Konwar |
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Institution: | 1. Division of Endocrinology, CSIR-Central Drug Research Institute (CDRI), U.P., Lucknow 226001, India;2. Division of Medicinal and Process Chemistry, CSIR-Central Drug Research Institute (CDRI), U.P., Lucknow 226001, India;3. Botany Division, CSIR-Central Drug Research Institute (CDRI), U.P., Lucknow 226001, India |
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Abstract: | Ethnopharmacological relevanceWrightia tomentosa Roem. & Schult. (Apocynaceae) is known in the traditional medicine for anti-cancer activity along with other broad indications like snake and scorpion bites, renal complications, menstrual disorders etc. However, the anti-cancer activity of this plant or its constituents has never been studied systematically in any cancer types so far.Aim of the studyTo evaluate the anti-cancer activities of the ethanolic extract of W. tomentosa and identified constituent active molecule(s) against breast cancer.Material and methodsPowdered leaves of W. tomentosa were extracted with ethanol. The ethanolic extract, subsequent hexane fractions and fraction F-4 of W. tomentosa were tested for its anti-proliferative and pro-apoptotic effects in breast cancer cells MCF-7 and MDA-MB-231.ResultsThe ethanolic extract, subsequent hexane fractions and fraction F-4 of W. tomentosa inhibited the proliferation of human breast cancer cell lines, MCF-7 and MDA-MB-231. The fraction F-4 obtained from hexane fraction inhibited proliferation of MCF-7 and MDA-MB-231 cells in concentration and time dependent manner with IC50 of 50 μg/ml and 30 μg/ml for 24 h, 28 μg/ml and 22 μg/ml for 48 h and 25 μg/ml and 20 μg/ml for 72 h respectively. The fraction F-4 induced G1 cell cycle arrest, reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential and subsequent apoptosis. Apoptosis is indicated in terms of increased Bax/Bcl-2 ratio, enhanced Annexin-V positivity, caspase 8 activation and DNA fragmentation. The active molecule isolated from fraction F-4, oleanolic acid and urosolic acid inhibited cell proliferation of MCF-7 and MDA-MB-231 cells at IC50 value of 7.5 μM and 7.0 μM respectively, whereas there is devoid of significant cell inhibiting activity in non-cancer originated cells, HEK-293. In both MCF-7 and MDA-MB-231, oleanolic acid and urosolic acid induced cell cycle arrest and apoptosis as indicated by significant increase in Annexin-V positive apoptotic cell counts.ConclusionOur results suggest that W. tomentosa extracts has significant anti-cancer activity against breast cancer cells due to induction of apoptosis pathway. Olenolic and urosolic acid are important constituent molecules in the extract responsible for anti-cancer activity of W. tomentosa. |
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Keywords: | ER Estrogen receptor DMEM Dulbecco's modified essential medium FCS Fetal calf serum DMSO Dimethyl sulphoxide PBS Phosphate buffer saline LDH Lactate dehydrogenase FITC Fluorescein isothiocyanate BrdU Bromodeoxyuridine qPCR Quantitative PCR AIF Apoptosis inducing factor ROS Reactive oxygen species FACS Fluorescence-activated cell sorter DCFH-DA-2&prime 7&prime Dichlorodihydrofluorescein diacetate DCF-2&prime 7&prime Dichlorofluorescein |
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