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Antiproliferative and
apoptotic effect of Astrodaucus orientalis (L.) drude
on T47D human breast
cancer cell line:
Potential mechanisms of action
M. H. Abdolmohammadi1,5*, S. Fouladdel1,2,
A. Shafiee3, G. Amin4, S. M. Ghaffari5
and E. Azizi1,2
1Department
of Pharmacology and Toxicology, Faculty of Pharmacy, Tehran
University of Medical Sciences, Tehran, Iran.
2National
Center of Excellence of Toxicology, Faculty of Pharmacy,
Tehran University of Medical Sciences, Tehran, Iran.
3Department
of Medicinal Chemistry and Pharmaceutical Sciences Research
Center, Faculty of Pharmacy, Tehran University of Medical
Sciences, Tehran, Iran.
4Department
of Pharmacognosy, Faculty of Pharmacy, Tehran University of
Medical Sciences, Faculty of Pharmacy, Tehran University of
Medical Sciences, Tehran, Iran.
5Institute
of Biochemistry and Biophysics (IBB), University of Tehran,
Tehran, Iran.
*Corresponding author. E-mail:
mh_abdolmohammadi@ibb.ut.ac.ir
Tel: +9821-66409517.
Fax:
+9821-66404680.
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; IC50, 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.
Accepted 3
April, 2009 |
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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.
Key
words:
Astrodaucus orientalis, T47D cells, breast cancer,
p53, Bcl-2, flowcytometry, RT-PCR, immunocytochemistry. |
