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Published on: September 19, 2018
Dissection of signaling pathways in fourteen breast cancer cell lines using reverse-phase protein lysate microarray
M Akkiprik1, D Nicorici, D Cogdell
1Department of Pathology, Unit 85, The University of Texas, M. D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030, USA.
Abstract:
Signal transduction pathways play a crucial role in breast cancer development, progression, and response to different therapies. A major problem in breast cancer therapy is the heterogeneity among different tumor types and cell lines commonly used in preclinical studies. To characterize the signaling pathways of some of the commonly used breast cancer cell lines and dissect the relationship among a number of pathways and some key genetic and molecular events in breast cancer development, such as p53 mutation, ErbB2 expression, and estrogen receptor (ER)/progesterone receptor (PR) status, we performed pathway profiling of 14 breast cancer cell lines by measuring the expression and phosphorylation status of 40 different cell signaling proteins with 53 specific antibodies using a protein lysate array. Cluster analysis of the expression data showed that there was close clustering of phosphatidylinositol 3-kinase, Akt, mammalian target of rapamycin (mTOR), Src, and platelet-derived growth factor receptor beta (PDGFRbeta) in all of the cell lines. The most differentially expressed proteins between ER- and PR-positive and ER- and PR-negative breast cells were mTOR, Akt (pThr308), PDGFRbeta, PDGFRbeta (pTyr751), panSrc, Akt (pSer473), insulin-like growth factor-binding protein 5 (IGFBP5), Src (pTyr418), mTOR (pSer2448), and IGFBP2. Many apoptotic proteins, such as apoptosis-inducing factor, IGFBP3, bad, bax, and cleaved caspase 9, were overexpressed in mutant p53-carrying breast cancer cells. Hexokinase isoenzyme 1, ND2, and c-kit were the most differentially expressed proteins in high and low ErbB2-expressing breast cancer cells. This study demonstrated that ER/PR status, ErbB2 expression, and p53 status are major molecules that impact downstream signaling pathways.
Insights
This study profiles signaling pathways in 14 breast cancer cell lines, revealing key protein differences linked to Estrogen Receptor/Progesterone Receptor status, ErbB2 expression, and p53 mutations. These findings clarify breast cancer heterogeneity and potential therapeutic targets.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Signal transduction pathways are critical in breast cancer development, progression, and therapy response.
- Breast cancer exhibits significant heterogeneity, complicating preclinical studies and treatment strategies.
- Understanding signaling pathway variations is essential for targeted breast cancer therapies.
Purpose of the Study:
- To characterize signaling pathways in common breast cancer cell lines.
- To investigate the relationships between pathways and key molecular events (p53 mutation, ErbB2 expression, ER/PR status).
- To identify differentially expressed proteins based on these molecular characteristics.
Main Methods:
- Pathway profiling of 14 breast cancer cell lines.
- Measurement of expression and phosphorylation of 40 cell signaling proteins using a protein lysate array with 53 antibodies.
- Cluster analysis of expression data.
Main Results:
- Phosphatidylinositol 3-kinase, Akt, mTOR, Src, and PDGFRbeta pathways showed close clustering across all cell lines.
- Significant protein expression differences were observed between ER/PR-positive and ER/PR-negative cells, including mTOR, Akt, PDGFRbeta, and Src.
- Mutant p53 breast cancer cells overexpressed apoptotic proteins, while ErbB2 expression levels correlated with Hexokinase isoenzyme 1, ND2, and c-kit.
- ER/PR status, ErbB2 expression, and p53 status were identified as major determinants of downstream signaling.
Conclusions:
- Breast cancer cell line heterogeneity impacts signaling pathway profiles.
- Specific signaling proteins and pathways are differentially regulated by Estrogen Receptor/Progesterone Receptor status, ErbB2 expression, and p53 mutation status.
- These molecular drivers significantly influence downstream signaling, offering insights into targeted therapeutic approaches.
