Time-dependent gene expression changes in a xenograft model of docetaxel cytotoxicity.

Abstract

Abstract Abstract #2009 Background: The antitumor effects of docetaxel primarily result from mitotic arrest with impaired cell proliferation. However, the key molecular pathways affected by this agent are largely unknown, and alternative mechanisms of action are likely. Knowledge of these pathways and mechanisms will guide the rational design of more effective treatment schedules and drug combinations. We hypothesize that the molecular analysis of treated human breast cancer xenografts will allow for an evaluation of time-dependent alterations in the expression of genes involved in key cell processes related to docetaxel cytotoxicity.
 Methods: Gene expression profiles were generated from MDA-MB-231 breast cancer xenografts after a single intravenous dose of docetaxel. Tumors were harvested at 1, 6, 12, and 24 hours (hr) after drug administration; a control group of untreated tumors (0hr) was included. RNA was extracted, assessed for concentration and quality, and used to generate probes for hybridization to the Affymetrix Human U133A oligonucleotide gene chip. Gene expression levels were explored with MAS 5.0 software and in-house algorithms. Subsequent data analysis included cross-experiment normalization, differential gene selection, gene clustering, and peak-time latency analysis. Consistency between the in-house in vivo dataset and a public in vitro dataset (Oncogene. 26:2902) was investigated.
 Results: 5087 responsive genes were selected using both SAM and EDGE. Among these genes, 3009 were down-regulated and 190 were up-regulated by at least 2-fold. Compared with 0h, the most significant changes in gene expression took place 6h after drug administration (2023 down-regulated and 655 up-regulated genes). Gene clustering revealed the sequence of up-regulation or down-regulation of different gene groups. Apoptosis-related genes and cell cycle-related genes in various stages showed different time latency patterns. For example, most of the M phase genes (total of 65 from Gene Ontology) were down-regulated 6h and 12h after docetaxel treatment and recovered by 24h. In addition, 1489 of 3009 down-regulated genes and 55 of 190 up-regulated genes were found in a public in vitro dataset; these datasets share 282 down-regulated genes (18.9%) and 17 up-regulated genes (30.9%).
 Discussion: Analysis of serial gene expression changes in docetaxel sensitive breast cancer xenografts demonstrates that time-dependent changes in specific cell processes occur. These findings will be expanded to distinguish between early and late molecular mechanisms of docetaxel responsiveness. A parallel xenograft model of docetaxel nonresponsiveness is also under investigation. In this way, time-dependent pathways will be defined that will include predictive biomarkers and lead to the development of novel agents to overcome resistance or enhance sensitivity to docetaxel, one of the most effective agents available for the treatment of breast cancer. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2009.