Determination of BRCA2 oncosuppressor protein expression in human mammary cells by affinity perfusion high-performance chromatography

Vissac, Cécile; Antoine-Vincent, David; Maurizis, Jean‐Claude; Bignon, Yves‐Jean; Bernard‐Gallon, Dominique

doi:10.1016/s0165-022x(01)00227-5

Cited by 7 publications

(3 citation statements)

References 12 publications

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“…We displayed the specificity of the anti-BRCA1 and anti-BRCA2 polyclonal antibodies by competition with the synthetic peptides used to generate the antibodies. A complete displacement of the equilibrium was obtained in each case demonstrating the specificity of the antibodies (data not shown) (Hizel et al, 1999;Vissac et al, 2001Vissac et al, , 2002.…”

Section: Discussionmentioning

confidence: 78%

Resveratrol increases BRCA1 and BRCA2 mRNA expression in breast tumour cell lines

et al. 2003

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The phytochemical resveratrol, found in grapes, berries and peanuts, has been found to possess cancer chemopreventive effects by inhibiting diverse cellular events associated with tumour initiation, promotion and progression. Resveratrol is also a phyto-oestrogen, binds to and activates oestrogen receptors that regulate the transcription of oestrogen-responsive target genes such as the breast cancer susceptibility genes BRCA1 and BRCA2. We investigated the effects of resveratrol on BRCA1 and BRCA2 expression in human breast cancer cell lines (MCF7, HBL 100 and MDA-MB 231) using quantitative real-time RT -PCR, and by perfusion chromatography of the proteins. All cell lines were treated with 30 mM resveratrol. The expressions of BRCA1 and BRCA2 mRNAs were increased although no change in the expression of the proteins were found. These data indicate that resveratrol at 30 mM can increase expression of genes involved in the aggressiveness of human breast tumour cell lines.

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Section: Discussionmentioning

confidence: 78%

Resveratrol increases BRCA1 and BRCA2 mRNA expression in breast tumour cell lines

et al. 2003

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“…The protein A affinity chromatography gives the amount of DNA-binding proteins that bind specifically to anti-BRCA1 or anti-BRCA2 antibodies, and a ratio is calculated as follows: (activity (disintegrations per minute) of BRCA1 or BRCA2 DNA-binding proteins that bind specifically to the anti-BRCA1 or BRCA2 antibodies : activity (disintegrations per minute) of labelled DNA-binding proteins eluted from heparin column) £100. All data are mean values and standard deviations of four assays (Hizel et al 1999;Vissac et al 2001). Untreated cells corresponded to the arbitrary value of 1 and treated cell expression was normalized to untreated cells.…”

Section: Brca1 and Brca2 Protein Quantificationmentioning

confidence: 99%

Differential effects ofn-3 andn-6 polyunsaturated fatty acids onBRCA1andBRCA2gene expression in breast cell lines

et al. 2002

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Current evidence strongly supports a role for the breast tumour suppressor genes, BRCA1 and BRCA2, in both normal development and carcinogenesis. In vitro observations reported that BRCA1 and BRCA2 are expressed in a cell cycle-dependent manner. Interestingly, differences in the actions of n-3 and n-6 polyunsaturated fatty acids have been observed: while the n-3 polyunsaturated fatty acids have been described to reduce pathological cell growth, the n-6 polyunsaturated fatty acids have been found to induce tumour proliferation. Here, we examined the expression of BRCA1 and BRCA2 in breast cell lines after treatment with polyunsaturated fatty acids. Real-time quantitative polymerase chain reaction determinations conclusively demonstrated increases in BRCA1 and BRCA2 mRNA expressions in MCF7 and MDA-MB 231 tumour cell lines after treatment with n-3 polyunsaturated fatty acids (eicosapentaenoic acid and docosahexaenoic acid), but no variation was noticed with the n-6 polyunsaturated fatty acid (arachidonic acid). On the other hand, no variation of the expression of BRCA1 and BRCA2 mRNA was detected in MCF10a normal breast cell line treated by polyunsaturated fatty acids. The level of BRCA1 and BRCA2 proteins quantified by affinity chromatography remained unchanged in tumour (MCF7, MDA-MB 231) and normal (MCF10a) breast cell lines. We suggest the presence of a possible transcriptional or post-transcriptional regulation of BRCA1 and BRCA2 after n-3 polyunsaturated fatty acid treatment in breast tumour cells.

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“…To accurately determine the expression level of BRCA1 or BRCA2 proteins in breast, a quantitative method previously developed by our laboratory to assay the expression of HLA-DR glycoproteins [23] was adapted to BRCA1 glycoproteins [24], late a faster method using perfusion chromatography was developed [25,26]. To measure the percentage of phosphorylated BRCA1 and BRCA2 compared with total proteins, three successive affinity chromatographies were used, starting with a heparin affinity column for initial isolation of DNA-binding proteins, because BRCA1 protein has a Zn finger at its amino terminus and BRCA2 has 8 BRC repeats, both characteristics of DNA-binding proteins.…”

Section: Introductionmentioning

confidence: 99%

Quantification by affinity perfusion chromatography of phosphorylated BRCAl and BRCA2 proteins from tumor cells after lycopene treatment

Chalabi

Maurizis

Corre

et al. 2005

Journal of Chromatography B

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Determination of BRCA2 oncosuppressor protein expression in human mammary cells by affinity perfusion high-performance chromatography

Cited by 7 publications

References 12 publications

Resveratrol increases BRCA1 and BRCA2 mRNA expression in breast tumour cell lines

Resveratrol increases BRCA1 and BRCA2 mRNA expression in breast tumour cell lines

Differential effects ofn-3 andn-6 polyunsaturated fatty acids onBRCA1andBRCA2gene expression in breast cell lines

Quantification by affinity perfusion chromatography of phosphorylated BRCAl and BRCA2 proteins from tumor cells after lycopene treatment

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