@article {16606,
	title = {Development and applications of a real-time quantitative RT-PCR method (QRT-PCR) for BRCA1 mRNA},
	journal = {Clin.Biochem.},
	volume = {38},
	year = {2005},
	note = {DA - 20041220
IS - 0009-9120 (Print)
IS - 0009-9120 (Linking)
LA - eng
PT - Journal Article
PT - Research Support, Non-U.S. Gov{\textquoteright}t
RN - 0 (Antineoplastic Agents)
RN - 0 (BRCA1 Protein)
RN - 0 (RNA, Messenger)
RN - EC 2.4.2.8 (Hypoxanthine Phosphoribosyltransferase)
SB - IM},
	month = {2005/01//},
	pages = {50 - 57},
	abstract = {OBJECTIVES: To develop a real-time quantitative RT-PCR method for BRCA1 mRNA and then use it for the study of BRCA1 gene expression in human MCF-7 breast cancer cells after their exposure to antineoplastic agents and gamma irradiation. DESIGN AND METHODS: The developed QRT-PCR method is based on the real-time monitoring of a fluorescein-labeled TaqMan probe, specific for BRCA1 mRNA, during PCR in the LightCycler. A BRCA1 PCR amplicon was purified, quantitated and used as a standard of known concentration for the development and analytical evaluation of the assay. The method was applied to study the alteration of BRCA1 gene expression after exposure to taxol, doxorubicin, 5-fluorouracil, etoposide or gamma irradiation in human MCF-7 breast cancer cells. RESULTS: The developed method is quantitative, highly specific for mRNA and highly sensitive (detection limit of 4 BRCA1 copies per mug of total RNA). We observed a reduction of BRCA1 expression for all antineoplastic agents used, while the gamma irradiated MCF-7 cells had an increase of expression with a peak at the 10 Gy dose. CONCLUSIONS: The developed BRCA1 QRT-PCR method is quantitative, highly sensitive and specific. The proposed method is rapid, automated, and cost effective and can be used to study BRCA1 expression in a variety of clinical samples},
	keywords = {Amino Acid Sequence, analysis, Antineoplastic Agents, Base Sequence, BRCA1 Protein, Breast Neoplasms, Carcinoma, Drug Therapy, Female, Gamma Rays, genetics, Greece, Humans, Hypoxanthine Phosphoribosyltransferase, metabolism, methods, Molecular Sequence Data, Reverse Transcriptase Polymerase Chain Reaction, RNA,Messenger, Tumor Cells,Cultured},
	url = {PM:15607317},
	author = {Kroupis, C. and Stathopoulou,A. and Zygalaki,E. and Ferekidou,L. and Talieri,M. and Lianidou,E.S.}
}
