Vissac Cécile, Peffault De Latour Monique, Communal Yves, Bignon Yves-Jean, Bernard-Gallon Dominique J
Laboratoire d'Oncologie Moléculaire, Centre Jean Perrin, 58 rue Montalembert-B.P. 392-63011, Clermont-Ferrand Cedex 1, France.
Clin Chim Acta. 2002 Jun;320(1-2):101-10. doi: 10.1016/s0009-8981(02)00055-4.
BRCA1 and BRCA2 are breast cancer susceptibility genes. Recent studies suggest that BRCA1 interacts with a great variety of proteins, including BRCA2, cell-cycle regulators, transcriptional activators and repressors. We investigated the expression of both BRCA1 and BRCA2 during the progression of the cell cycle of human tumor cell lines from different origins (MCF7, MDA-MB231, PA1 and CCL221) in two growth status (60% and 100% of confluency).
First, the growth status was characterized by determination of the cell cycle by flow cytometry analysis. At the same time, immunohistochemistry was performed to follow BRCA1 and BRCA2 protein expression and then, quantification of BRCA1 and BRCA2 transcripts was realized using real-time quantitative RT-PCR.
We reported in studied tumor cell lines with 60% of confluency by comparison with 100% of confluency, an increase in the BRCA1 and BRCA2 expression at the level of proteins and transcripts.
Therefore, the expression of both BRCA1 and BRCA2 genes at the protein and mRNA levels appear to be up-regulated after cell proliferation in human tumor cell lines from different origins.
BRCA1和BRCA2是乳腺癌易感基因。最近的研究表明,BRCA1与多种蛋白质相互作用,包括BRCA2、细胞周期调节因子、转录激活因子和抑制因子。我们研究了来自不同来源的人类肿瘤细胞系(MCF7、MDA-MB231、PA1和CCL221)在两种生长状态(60%和100%汇合度)的细胞周期进程中BRCA1和BRCA2的表达情况。
首先,通过流式细胞术分析确定细胞周期来表征生长状态。同时,进行免疫组织化学以追踪BRCA1和BRCA2蛋白表达,然后使用实时定量RT-PCR实现BRCA1和BRCA2转录本的定量分析。
我们报道,在所研究的肿瘤细胞系中,与100%汇合度相比,60%汇合度时BRCA1和BRCA2在蛋白质和转录本水平的表达增加。
因此,在来自不同来源的人类肿瘤细胞系中,细胞增殖后BRCA1和BRCA2基因在蛋白质和mRNA水平的表达似乎均上调。