Related Experiment Video
Updated: Feb 1, 2026
Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells
Published on: April 14, 2010
Differential Gene Expression in Erlotinib-Treated Fibroblasts
Karen E Wickersham1, Theresa K Hodges, Martin J Edelman
1Karen E. Wickersham, PhD, RN, was Assistant Professor, School of Nursing, University of Maryland, Baltimore; now Assistant Professor, University of South Carolina, College of Nursing, Columbia, South Carolina. Theresa K. Hodges, PhD, is Bioinformatics Analyst I, Institute for Genome Sciences, School of Medicine, University of Maryland, Baltimore, Maryland. Martin J. Edelman, MD, was Director, Medical Thoracic Oncology, University of Maryland Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland; now Professor and Chair, Department of Hematology/Oncology; Deputy Director, Cancer Center for Clinical Research; and G. Morris Dorrance Jr. Chair in Medical Oncology, Fox Chase Cancer Center, Philadelphia, Pennsylvania. Yang Song, PhD, is Bioinformatics Analyst II, Institute for Genome Sciences, School of Medicine, University of Maryland, Baltimore, Maryland. Mintong Nan, BS, was Laboratory Research Technician, Department of Pain and Translational Symptom Science, University of Maryland, Baltimore, School of Nursing, Baltimore, Maryland. Susan G. Dorsey, PhD, RN, FAAN, is Professor and Chair, Department of Pain and Translational Symptom Science, University of Maryland, Baltimore, School of Nursing, Baltimore, Maryland; and PhD Student at the University of Maryland.
Background:
Therapies targeting the epidermal growth factor receptor (EGFR) result in a painful rash, the most common and debilitating toxicity among patients with non-small cell lung cancer (NSCLC) who take EGFR tyrosine kinase inhibitor (TKI) therapy; however, predicting the development and the severity of the rash is difficult.
Objective:
The aim of this study was to examine how erlotinib-an EGFR TKI that NSCLC patients take to stop or slow tumor growth-altered the transcriptome of dermal fibroblasts.
Methods:
Dermal fibroblasts (ATCC PCS-201-012) were seeded in cell culture flasks, grown under standard conditions, and transferred to cell culture dishes. Cells were treated once daily for 3 days with erlotinib 100 nM (n = 5), erlotinib 1 μM (n = 5), vehicle 1 μM (dimethyl sulfoxide) (n = 5), or no treatment (n = 5). Total RNA was extracted using a standard TRIzol method and hybridized using Affymetrix GeneChip Human Genome U133 Plus 2.0 arrays. Raw intensities generated from the arrays were normalized using a Robust Multiarray Average method and analyzed using analysis of variance in Limma R software. Differentially expressed genes were analyzed using Ingenuity Pathway Analysis to identify canonical or noncanonical signaling pathways enriched in this dataset.
Results:
We selected genes for investigation based on their potential role in wound healing (AQP3), rash development (CCL2), fibroblast activation (PALLD), cancer and cancer progression (GDF-15, SLC7A11, MMP12, and DIRAS3), and cell cycle control (CDC6). We were able to validate four of these genes by both Western blot analysis and quantitative polymerase chain reaction (MMP12, CCL2, CDC6, and SLC7A11).
Discussion:
If found predictive of rash in future studies using patient samples, our findings may help to identify those at risk for severe rash so that (a) the dose of EGFR TKI therapy may be adjusted; (b) additional treatments for the rash can be developed; and/or (c) precise, patient-centered interventions can be developed so that patients with cancer can better self-manage their rash and adhere to EGFR TKI treatment.
Insights
Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) therapy for non-small cell lung cancer can cause severe skin rash. This study identified specific genes in dermal fibroblasts altered by erlotinib, potentially predicting rash development.
Area of Science:
- Dermatology
- Oncology
- Molecular Biology
Background:
- Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) therapies are crucial for non-small cell lung cancer (NSCLC) treatment.
- A common and debilitating side effect of EGFR-TKI therapy is a painful skin rash, making prediction and management challenging.
Purpose of the Study:
- To investigate the transcriptomic changes in dermal fibroblasts induced by erlotinib, an EGFR-TKI.
- To identify potential biomarkers for predicting EGFR-TKI-induced rash in NSCLC patients.
Main Methods:
- Dermal fibroblasts were treated with varying concentrations of erlotinib or vehicle control.
- Gene expression profiling was performed using Affymetrix arrays.
- Differentially expressed genes were analyzed using Ingenuity Pathway Analysis.
Main Results:
- Erlotinib treatment altered the expression of several genes in dermal fibroblasts.
- Key genes related to wound healing, inflammation, and cancer progression were identified.
- Four genes (MMP12, CCL2, CDC6, and SLC7A11) were validated using Western blot and qPCR.
Conclusions:
- The identified gene expression changes may serve as predictive markers for EGFR-TKI-induced rash.
- Findings could lead to better patient risk stratification and personalized treatment strategies.
- Potential for developing targeted interventions to manage rash and improve treatment adherence.
Related Concept Videos
01:42What is Gene Expression?
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
01:36What is Gene Expression?
02:35Chromatin Position Affects Gene Expression
Topologically Associated Domains (TADs)
The 3-dimensional positioning of chromatin in the nucleus influences the...
01:58Cell Specific Gene Expression
01:58Cell Specific Gene Expression
02:51mRNA Stability and Gene Expression
Cis-acting Elements involved in mRNA stability