紫杉醇诱导的卵巢癌耐药基因表达谱与信号通路分析

The Analysis of Gene Expression Profiling and Network of Paclitaxel-resistant Cell Line in Ovarian Cancer

目的:探讨卵巢癌细胞对紫杉醇的耐药分子机制。方法:取对数生长期细胞按Trizol一步法抽提细胞总RNA,应用人类全基因组基因芯片检测卵巢癌耐药细胞株SKOV3/TS与敏感细胞株SKOV3细胞基因表达谱的变化,GOTermMapper软件鉴定差异基因功能,进一步利用IPA软件分析基因通路网络,筛查获得性紫杉醇卵巢癌耐药的相关基因。结果:紫杉醇诱导的卵巢癌耐药差异表达基因分别与细胞生长增殖,细胞合成和装配,细胞死亡,细胞周期调控和细胞信号传导相关。涉及DNA修复功能,DNA复制和细胞周期阻滞的基因主要是过度表达,而在代谢相关基因(特别是脂质代谢),信号传导,免疫和炎症反应,运输,转录调控和蛋白质的合成则是抑制表达。网络分析结果表明微管蛋白网络和以BAX为节点的细胞凋亡信号转导通路被促进。结论:紫杉醇耐药主要是与细胞合成与装备(包括许多微管蛋白基因诱导相关的基因和通路),脂质代谢、细胞粘附和细胞凋亡相关。BAX蛋白从微管蛋白释放被抑制可能是诱导卵巢癌细胞耐药的重要原因。

Objective： The mechanism of chemotherapy-resistance in ovarian cancer is complex. To better understand the molecular mechanisms of ovarian cancer cells to paclitaxel exposure, we have performed gene expression proffling in paclitaxel-resistant cell line SKOV3/TS by comparing it with its parental cell SKOV3 and to explore the relationship between differential gene expression and drug-resistance. Methods： Total RNA was extracted from the cell line during the log phase with Trizol one-step method. Whole human genome microarray which included all annotated human genes （32 050 probe sets） was used. The differential genes compared between the SKOV3 3/TS and SKOV3 cell line were classified according to their function using the Gene Ontology （ GO TermMapper） classification system. Network analysis of the microarray data was completed using the Ingenuity Pathway Analysis software. Results： The differential expressions of gene induced by paclitaxel were associated with cell growth and proliferation, cellular assembly and organization, cell death, cell cycle control and cell signaling. Genes, functionally involved in DNA repair, DNA replication and cell cycle arrest were mostly over expressed, while genes implicated in metabolism （ especially lipid metabolism）, signal transduction, immune and inflammatory response, transport, transcription regulation and protein biosynthesis were commonly suppressed. Network analysis results showed that the tubulin network and BAX mediated apoptosis signal transduction pathway were promoted. Conclusion： The drug-resistance of paclitaxel is mainly associated with induction of genes and pathways linked to cellular assembly and organization（ including numerous tubul in genes ）, lipid metabolism, cell adhesion and cell apoptosis. Our study demonstrates that the inhibited release of BAX protein from tubulin may be a major cause of paclitaxel -resistance in ovarian cancer.