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Antiproliferative activity and standardization of Tecomella undulata bark extract on K562 cells
Authors:Ravi Alvala  Mallika A  Sama Venkatesh  Begum A Sajeli  Khan Rukaiyya S  Reddy B Madhava
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
Abstract:

Ethnopharmacological relevance

The 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.

Aim

To 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.

Methods

Induction 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.

Result

Results 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.

Conclusion

The 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.
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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