Antiproliferative activity and standardization of Tecomella undulata bark extract on K562 cells |
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Authors: | Ravi Alvala Mallika A Sama Venkatesh Begum A Sajeli Khan Rukaiyya S Reddy B Madhava |
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Affiliation: | a G. Pulla Reddy College of Pharmacy, Mehdipatnam, Hyderabad-500 028, AP, India b Birla Institute of Technology and Science, Pilani-Hyderabad Campus, Hyderabad-500 078, AP, India |
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Abstract: | Ethnopharmacological relevanceThe bark of Tecomella undulata is primarily used in the treatment of syphilis, painful swellings and cancer by traditional healers. Also, it is claimed to be useful in treating urinary discharges, enlargement of spleen, leucorrhoea, leukoderma, tumors, liver disorders, gonorrhea, gout and promotes wound healing in Indian traditional system of medicine.AimTo establish a scientific validation for the antitumor effects of Tecomella undulata bark and explore the mechanistic pathway in chronic myeloid leukemia cell line, K562. The study was further extended to standardize the extract using quercetin as biomarker.MethodsInduction of apoptosis by chloroform extract of Tecomella undulata bark (CTUB) was determined by MTT, Annexin V and caspase activation assays. The cell cycle analysis was done by flow cytometer and nuclear staining by DAPI. The standardization of the extract was performed through reverse phase-HPLC method under PDA detection.ResultResults clearly showed the induction of apoptosis by CTUB in K562 cells. The effect was found to be dose dependent, having IC50 of 30 μg/ml with activation of FAS, FADD, caspase 8, caspase 3/7 and fragmentation of DNA. The bioactive CTUB was determined to possess 0.03% (w/w) of quercetin.ConclusionThe investigation clearly demonstrated the potential antitumor effect of CTUB, thereby validating the traditional claim. Quercetin, known to have anticancer activity is being reported and quantified for the first time from the bark of Tecomella undulata. |
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Keywords: | 7-AAD, 7-aminoactinomycin D ANOVA, one-way analysis of variance Caspase, cysteine-aspartic proteases or cysteine-dependent aspartate-directed proteases COLO-205, human colon carcinoma cell CTUB, chloroform extract of Tecomella undulata bark DAPI, 4&prime ,6-diamidino-2-phenylindole DMSO, dimethyl sulfoxide FADD, Fas-associated protein with death domain FAS, TNF receptor super familymember 6 FBS, fetal bovine serum FLICA, Fluorescent Labeled Inhibitor of Caspases HepG2, human hepatocellular liver carcinoma cell line HPLC, high-performance liquid chromatography K562, chronic myeloid leukemia MDA-MB23, human breast carcinoma cells MTT, 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide PBS, phosphate-buffered saline PDA, Photo Diode Array PI, propidium iodide PS, phosphatidylserine RPMI, Roswell Park Memorial Institute |
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