Abstract:
Retinoblastoma (RB) is a pediatric cancer of the eye that occurs in 1/15000 live births worldwide. Albeit RB is initiated by the inactivation of RB1 gene, the disease progression relies largely on transcriptional alterations. Therefore, evaluating gene expression is vital to unveil the therapeutic targets in RB management. In this study, we employed an RT2 Profiler™ PCR array for a focused analysis of 84 cancer-specific genes in RB. An interaction network was built with gene expression data to identify the dysregulated pathways in RB. The key transcript alterations identified in 13 tumors by RT2 Profiler™ PCR array was further validated in 15 tumors by independent RT-qPCR. Out of 84 cancer-specific genes, 68 were dysregulated in RB tumors. Among the 68 genes, 23 were chosen for further analysis based on statistical significance and abundance across multiple tumors. Pathway analysis of altered genes showed the frequent perturbations of cell cycle, angiogenesis and apoptotic pathways in RB. Notably, upregulation of MCM2, MKI67, PGF, WEE1, CDC20 and downregulation of COX5A were found in all the tumors. Western blot confirmed the dysregulation of identified targets at protein levels as well. These alterations were more prominent in invasive RB, correlating with the disease pathogenesis. Our molecular analysis thus identified the potential therapeutic targets for improving retinoblastoma treatment. We also suggest that PCR array can be used as a tool for rapid and cost-effective gene expression analysis.
摘要:
视网膜母细胞瘤(RB)是一种儿科眼癌,发生在全球1/15000活产中。尽管RB是由RB1基因失活引发的,疾病进展在很大程度上依赖于转录改变。因此,评估基因表达对于揭示RB管理中的治疗靶标至关重要。在这项研究中,我们使用RT2Profiler™PCR阵列对RB中84个癌症特异性基因进行了集中分析.利用基因表达数据构建相互作用网络以鉴定RB中失调的途径。通过RT2Profiler™PCR阵列在13种肿瘤中鉴定的关键转录物改变通过独立的RT-qPCR在15种肿瘤中进一步验证。在84个癌症特异性基因中,68在RB肿瘤中失调。在68个基因中,基于多个肿瘤的统计显著性和丰度,选择23个用于进一步分析。改变基因的通路分析显示细胞周期的频繁扰动,RB中的血管生成和凋亡途径。值得注意的是,MCM2、MKI67、PGF、在所有肿瘤中均发现了WEE1,CDC20和COX5A的下调。Western印迹也证实了所鉴定的靶标在蛋白质水平上的失调。这些改变在侵入性RB中更为突出,与疾病的发病机制有关。因此,我们的分子分析确定了改善视网膜母细胞瘤治疗的潜在治疗靶点。我们还建议PCR阵列可用作快速且具有成本效益的基因表达分析工具。