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The Role of CtBP in Tumor Progression: Insights from the pINDUCER inducible gene expression system
Author(s) -
Yi Dae Ik,
Li Daniel,
Park Sam,
Crawford Lindsey,
Liang Genqing,
Kabbout Mohamed,
HernandezMora Roberto,
Yan Tingfen,
Byun Jung,
Gardner Kevin
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.698.11
Subject(s) - carcinogenesis , tumor progression , cancer research , biology , epigenetics , tumor initiation , metastasis , microrna , corepressor , transcription factor , epithelial–mesenchymal transition , psychological repression , regulation of gene expression , gene expression , genome instability , chromatin , microbiology and biotechnology , gene , cancer , genetics , dna damage , dna
As a transcriptional corepressor, CtBP plays a significant role in tumorigenesis and tumor progression through epigenetic regulation. Interacting with transcription factors and chromatin modifying enzymes, CtBP mediates repression of tumor suppressor genes that maintain genome stability, repress cellular proliferation and inhibit EMT. This contributes to the evolution of more aggressive forms of multiple epithelial malignancies, including breast cancer. CtBP is also considered a metabolic sensor due to its ability to bind to NADH, the high energy intermediate generated during carbohydrate metabolism. However, despite its potential role in tumor progression and its link to metabolic imbalance, a detailed mechanism explaining how CtBP promotes aggressive breast cancer behavior is unclear. Thus, we are employing an inducible gene expression approach that utilizes the lentiviral pINDUCER system to temporally manipulate the levels of CtBPs using Doxycycline. We successfully established three inducible human mammary epithelial cell lines that exhibit distinct characteristics spanning from Luminal A to Triple‐Negative, Claudin‐low type such as MDA‐MB‐231, MCF7, and MCF10A. The fidelity of this system allows us to characterize CtBP‐regulated pathways including EMT, invasion, metastasis, genome instability, and uncontrolled self‐renewal. Functional assays such as radius migration and Boyden chamber invasion assays showed that high expression of CtBP correlated with mesenchymal features including EMT. On the other hand, low CtBP expression resulted in epithelial‐like characteristics. Through metabolomics studies, we also found that CtBP drives deregulated cellular energetics such as Warburg‐like effects in these mammary epithelial cell lines. Eventually, we will develop orthotopic xenograft mouse models to study breast cancers in vivo. Together, this functional implementation of the pINDUCER system will shed light on the role of CtBP in breast tumorigenesis and progression. Support or Funding Information NCI, NIHDiagrams of the pINDUCER vector seriespINDUCER10‐14: CtBP Knockdown / pINDUCER20–22: CtBP OverexpressionComparison of cell motility and wound healing process in MDA‐MB‐231 to check how CtBP expression influences EMT or aggressive behaviors of breast cancer