Antiproliferative and apoptotic effect of Astrodaucus orientalis (L.) drude on T47D human breast cancer cell line: Potential mechanisms of action

Abbreviations: AEC, 3-Amino-9-ethylcarbazole; ATCC, American type culture collection; DAPI, 4, 6-diamidine-2-phenylindole dihydrochloride; DEPC, diethylpyrocarbonate; DMSO, dimethyl sulfoxide; dNTP, PCR nucleotide mix; DTT, dithiothreitol; FBS, fetal bovine serum; FITC, fluorescein isothiocyanate; IC sub(50), the median inhibitory concentration; M-MuLV, reverse transcriptase; MTT, 3-(4,5-Dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide; PBS, phosphate buffer solution; Pen-strep, penicillin-streptomycin; PI, propidium iodide; TAM, tamoxifen; triPure, RNA, DNA and protein isolation reagent. Breast cancer is one of the most common cancers in women. Here, we report the antiproliferative and the mechanism of cell death exhibited by Astrodaucus orientalis in human breast carcinoma cell line, T47D. The antiproliferative effect determined with MTT assay and the changes in cell cycle pattern were studied with DAPI reagent and Annexin V/PI method used for apoptotic analysis. The p53 and Bcl-2 are believed to play a crucial role in tumorigenesis and cell death. The p53 and Bcl-2 genes and proteins expression alterations in T47D were studied using RT-PCR and immunocytochemistry technique. Both extracts showed strong antiproliferative effects on cells. Root extract showed relatively similar cell cycle pattern to RPMI but the aerial extract showed cell cycle arrest in G2/M. Both extracts induced apoptosis and the effect of aerial extract was more prominent than that of root extract. The p53 gene expression decreased in presence of extracts and Bcl-2 gene expression also significantly decreased as their expressions had not detectable. Both extracts also showed decrease in p53 and Bcl-2 protein expression. These data are first reported on potential of anticancer activity of A. orientalis extracts and one of its possible mechanisms of action on cancer cell proliferation.