Related Experiment Video
Updated: May 31, 2026
Generation of Prostate Cancer Cell Models of Resistance to the Anti-mitotic Agent Docetaxel
Published on: September 8, 2017
Two different docetaxel resistant MCF-7 sublines exhibited different gene expression pattern
Ozlem Darcansoy Işeri1, Meltem Demirel Kars, Ufuk Gündüz
1Department of Biological Sciences, Middle East Technical University, 06531, Ankara, Turkey.
Abstract:
The objective of the present study was to investigate gene expression pattern of two docetaxel resistant MCF-7 breast carcinoma sublines step wisely selected in 30 and 120 nM docetaxel. Cell proliferation assay was performed in order to demonstrate development of docetaxel resistance. cDNA microarray analysis was performed using Affymetrix(®) Human Genome U133 Plus 2.0 Arrays in duplicate experiments. Quantitative and semi-quantitative gene expression analysis was also performed to confirm gene expression analysis for selected genes. XTT results demonstrated that 30 (MCF-7/30nM DOC) and 120 nM (MCF-7/120nM DOC) docetaxel selected cells were 13- and 47-fold resistant, respectively. cDNA microarray analysis demonstrated that expression profiles of MCF-7 and MCF-7/30nM DOC were more similar to each other where expression profile of MCF-7/120nM DOC was different as examined by line graphs and scatter plots. 2,837 and 4,036 genes were significantly altered in 30 and 120 nM docetaxel resistant sublines, respectively. Among these, 849 genes were altered in common in two docetaxel resistant sublines. Antiapoptotic gene expression (e.g., Bcl-2 and APRIL) were noticeably altered in MCF-7/30nM DOC. However, docetaxel resistance in MCF-7/120nM DOC were more complicated with the involvement of ECM related gene expression, cytokine and growth factor signaling, ROS metabolism and EMT related gene expression together with higher level of MDR1 expression. Expression profiles in 30 and 120 nM docetaxel resistant sublines changed gradually with increasing resistance index. Drug resistance development seems to be step wise event in MCF-7 cells.
Insights
This study reveals that docetaxel resistance in MCF-7 breast cancer cells develops stepwise. Gene expression changes, including anti-apoptotic and EMT-related genes, increase with docetaxel resistance levels.
Area of Science:
- Oncology
- Molecular Biology
- Genomics
Background:
- Docetaxel is a key chemotherapy agent for breast cancer.
- Acquired drug resistance limits treatment efficacy.
- Understanding resistance mechanisms is crucial for improving patient outcomes.
Purpose of the Study:
- To investigate gene expression patterns in docetaxel-resistant MCF-7 breast carcinoma sublines.
- To characterize the stepwise development of docetaxel resistance.
- To identify molecular markers associated with increasing resistance levels.
Main Methods:
- Development of docetaxel-resistant MCF-7 sublines (MCF-7/30nM DOC and MCF-7/120nM DOC).
- Cell proliferation assays (XTT) to quantify resistance.
- cDNA microarray analysis (Affymetrix Human Genome U133 Plus 2.0 Arrays).
- Quantitative and semi-quantitative gene expression analysis.
Main Results:
- MCF-7/30nM DOC and MCF-7/120nM DOC exhibited 13- and 47-fold docetaxel resistance, respectively.
- Gene expression profiles diverged with increasing resistance; MCF-7/120nM DOC showed significant alterations.
- 2,837 and 4,036 genes were altered in 30nM and 120nM resistant sublines, respectively.
- Antiapoptotic genes (Bcl-2, APRIL) were altered in MCF-7/30nM DOC.
- MCF-7/120nM DOC resistance involved ECM, cytokine signaling, ROS metabolism, EMT, and MDR1 expression.
Conclusions:
- Docetaxel resistance in MCF-7 cells is a gradual, stepwise event.
- Distinct molecular mechanisms contribute to early-stage (30nM) versus advanced (120nM) docetaxel resistance.
- Gene expression profiling provides insights into the complex pathways underlying acquired chemoresistance.
