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Proteomic analysis of estrogen response of premalignant human breast cells using a 2‐D liquid separation/mass mapping technique
Author(s) -
Zhao Jia,
Zhu Kan,
Lubman David M.,
Miller Fred R.,
Shekhar Malthy P. V.,
Gerard Brigitte,
Barder Timothy J.
Publication year - 2006
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500195
Subject(s) - estrogen , chromatofocusing , proteomics , estrogen receptor , chemistry , phenotype , biology , microbiology and biotechnology , biochemistry , enzyme , breast cancer , cancer , isoelectric point , endocrinology , gene , genetics
A 2‐D liquid‐phase separation method based on chromatofocusing and nonporous silica RP‐HPLC followed by ESI‐TOF‐MS was used to analyze proteins in whole cell lysates from estrogen‐treated and untreated premalignant, estrogen‐responsive cell line MCF10AT1 cells. 2‐D mass maps in the pH range 4.6–6.0 were generated with good correlation to theoretical M r values for intact proteins. Proteins were identified based on intact M r , p I and PMF, or MS/MS sequencing. About 300 unique proteins were identified and 120 proteins in mass range 5–75 kDa were quantified upon treatment of estrogen. Around 40 proteins were found to be more highly expressed (>four‐fold) and 17 were down‐regulated (>four‐fold) in treated cells. In our study, we found that many altered proteins have characteristics consistent with the development of a malignant phenotype. Some of them have a role in the ras pathway or play an important role in signal pathways. These changed proteins might be essential in the estrogen regulation mechanism. Our study highlights the use of the MCF10AT1 cell line to examine estrogen‐induced changes in premalignant breast cells and the ability of the 2‐D mass mapping technique to quantitatively study protein expression changes on a proteomic scale.