Khabele Dineo, Lopez-Jones Melissa, Yang WanCai, Arango Diego, Gross Susan J, Augenlicht Leonard H, Goldberg Gary L
Albert Einstein Cancer Center and Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Gynecol Oncol. 2004 Apr;93(1):19-26. doi: 10.1016/j.ygyno.2003.11.058.
Epothilone B (EpoB) is a non-taxane microtubule-stabilizing agent with a mode of action similar to that of paclitaxel, but with the advantage of being active in paclitaxel-resistant cells. Knowledge regarding other mechanisms of EpoB action is limited. The purpose of this study was to identify gene expression profiles associated with the biological response to EpoB in an ovarian cancer cell line (SKOV3).
SKOV3 cells were maintained in McCoy's 5A media. Equal densities cells were treated with or without EpoB, and were evaluated for cell growth arrest and apoptosis. mRNA expression was evaluated by cDNA microarrays and quantitative, real time reverse transcription polymerase chain reaction (QRTPCR).
EpoB (10 nM) led to cell cycle arrest and apoptosis in SKOV3 cells. Microarray analysis, comparing EpoB-treated to untreated cells, revealed altered expression of 41 genes. There was a predominance of sequences related to the TNFalpha stress response pathway. Differential expression of selected genes was confirmed by QRTPCR.
We demonstrated that cDNA microarrays are a useful tool to rapidly screen for patterns of gene expression that characterize drug response. The microarray data suggest that the microtubule-stabilizing agent, EpoB, triggers stress-related signal transduction pathways related to TNFalpha. These pathways may contribute to mechanisms of EpoB action and potential mechanisms of resistance in ovarian cancer.
埃坡霉素B（EpoB）是一种非紫杉烷类微管稳定剂，其作用模式与紫杉醇相似，但具有在紫杉醇耐药细胞中仍保持活性的优势。关于EpoB作用的其他机制的知识有限。本研究的目的是确定与卵巢癌细胞系（SKOV3）对EpoB的生物学反应相关的基因表达谱。
SKOV3细胞在 McCoy's 5A培养基中培养。将等密度的细胞用或不用EpoB处理，并评估细胞生长停滞和凋亡情况。通过cDNA微阵列和定量实时逆转录聚合酶链反应（QRTPCR）评估mRNA表达。
EpoB（10 nM）导致SKOV3细胞的细胞周期停滞和凋亡。微阵列分析比较了用EpoB处理的细胞和未处理的细胞，发现41个基因的表达发生了改变。与肿瘤坏死因子α应激反应途径相关的序列占主导。通过QRTPCR证实了所选基因的差异表达。
我们证明cDNA微阵列是快速筛选表征药物反应的基因表达模式的有用工具。微阵列数据表明，微管稳定剂EpoB触发了与肿瘤坏死因子α相关的应激相关信号转导途径。这些途径可能有助于EpoB的作用机制以及卵巢癌耐药的潜在机制。
