Gene expression profiling of a clonal isolate of oxaliplatin-resistant ovarian carcinoma cell line A2780/C10.
Gene expression profiling of a clonal isolate of oxaliplatin-resistant ovarian carcinoma cell line A2780/C10.
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DOI：
10.3892/or.14.4.925
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发表时间：
2005-10
期刊：
影响因子：
4.2
通讯作者：
R. Varma;S. Hector;K. Clark;W. Greco;L. Hawthorn;L. Pendyala
中科院分区：
文献类型：
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作者：
R. Varma;S. Hector;K. Clark;W. Greco;L. Hawthorn;L. Pendyala
铂类药物在癌症治疗中的疗效通常受到治疗后肿瘤细胞耐药性的限制。为了更好地理解这种现象的机制，如前所述，从奥沙利铂耐药的人卵巢癌细胞系（A2780/C10）中分离出一个克隆亚系（A2780/C10 B）。该细胞系对奥沙利铂的耐药性为18倍，对顺铂的交叉耐药性为3倍。在这里，我们报告了使用来自亲本细胞系和耐药变异体的对数生长期细胞的Affyphase HG-U95 Av 2寡核苷酸阵列的基因表达分析。使用基于模型的表达指数（dChip）和稳健多芯片平均（RMA）方法进行探针水平分析。采用微阵列显著性分析（SAM）方法鉴定两组间差异表达的基因，最小错误发现率<1%。我们在耐药细胞系中发现了43个过度表达的基因和39个低表达的基因。胶原VI（COL 6A 3）过表达62倍，是最高度上调的基因。这一发现与其他基于顺铂耐药和敏感卵巢癌细胞基因表达谱系列分析（SAGE）的已发表数据一致。在我们的研究中，过表达基因的重要功能组是细胞外基质基因（43个中的9个）和参与信号转导的基因（43个中的7个）。细胞外基质基因包括两种基质金属蛋白酶（MMP 3和MMP 12）。整合素α 1（ITGA 1）和WNT 5A也过表达。编码细胞外基质蛋白的基因也在耐药细胞系中被发现下调。发现参与细胞周期和生长调节的几个基因表达不足，包括细胞因子信号传导抑制因子2（SOCS 2）、necdin（NDN）和磷脂酰肌醇蛋白聚糖（GPC 3和GPC 4）。采用实时荧光定量RT-PCR方法对6个差异表达基因（COL 6A 3、MMP 12、MMP 3、WNT 5A、NID和HMGB 2）的mRNA水平进行验证。这些基因的鉴定应该有助于更好地了解导致铂类药物耐药性的途径。
The efficacy of platinum drugs in the treatment of cancer is often restricted by the acquisition of tumor cell resistance subsequent to treatment. To better understand mechanisms involved in this phenomenon, a clonal subline (A2780/C10B) isolated from an oxaliplatin-resistant human ovarian carcinoma cell line (A2780/C10) was developed, as reported previously. This cell line is 18-fold resistant to oxaliplatin and shows a 3-fold cross resistance to cisplatin. Here, we report on the gene expression analysis using Affymetrix HG-U95Av2 oligonucleotide arrays of cells in log phase growth from both the parental cell line and drug-resistant variant. Probe level analysis was perfomed using the model based expression index (dChip) and robust multichip average (RMA) methods. Genes that were differentially expressed between the two groups were identified using the significance analysis of microarrays (SAM) method with a minimum false discovery rate <1%. We identified 43 genes that were overexpressed, and 39 underexpressed in the drug-resistant cell line. Collagen VI (COL6A3) was overexpressed 62-fold and the most highly up-regulated gene. This finding is consistent with other published data based on serial analysis of gene expression (SAGE) profiling of cisplatin-resistant and sensitive ovarian carcinoma cells. Among the significant functional groups of overexpressed genes in our study were extracellular matrix genes (9 of 43) and those involved in signal transduction (7 of 43). Extracellular matrix genes included two matrix metalloproteinases (MMP3 and MMP12). Integrin alpha 1 (ITGA1) and WNT5A were also overexpressed. Genes that encode for extracellular matrix proteins were also among those found down-regulated in the resistant cell line. Several genes involved in the regulation of cell cycle and growth were found to be underexpressed, including the suppressor of cytokine signaling 2 (SOCS2), necdin (NDN), and glypicans (GPC3 and GPC4). The mRNA levels of six differentially expressed genes (COL6A3, MMP12, MMP3, WNT5A, NID, and HMGB2) were validated using real-time quantitative RT-PCR. The identification of these genes should aid in a better understanding of the pathways resulting in platinum drug resistance.