背景:下咽癌是一种相对罕见的恶性肿瘤,预后较差。目前的化疗算法离个性化医疗还很远,而真正有活性的治疗性生物标志物和/或靶标的鉴定是热切期待的。
方法:关注常规关键化疗药物,我们确定了与多西他赛(TXT)细胞敏感性最相关的基因(和/或蛋白质),顺铂(CDDP)和5-氟尿嘧啶(5-FU)的表达水平,通过3个步骤的方法:全基因组筛选,关于量化表达水平的确认研究,以及候选物的敲低和转染分析。所选基因的可能作用途径通过使用大规模数据库的创造性途径分析进行了检查,癌症基因组图谱。
结果:首次全基因组筛选研究在15个细胞系中获得了16个与细胞药物敏感性高度相关的基因(CDDP和5-FU的|R|>0.8,P<0.01;TXT的|R|>0.5,P<0.05)。在10个基因中,观察到的相关性在定量的基因表达水平上得到了证实,最后,敲除和转染分析提供了4个分子作为最有效的预测标记-AGR2(前梯度2同源基因),和PDE4D(磷酸二酯酶4D,cAMP特异性基因)用于TXT;NINJ2(神经损伤诱导蛋白2);CDC25B(细胞分裂周期25同源物B基因)用于5-FU-在基因和蛋白质表达水平上。AGR2、PDE4D的过表达表明对TXT的反应较差,抑制的表达使TXT活性敏感。与调查结果相反,在其他两个分子中,NINJ2和CDC25观察到与细胞药物对相关药物的反应相反的关系。IPA提高了每种选择的分子与相关药物的主要作用途径(和/或靶标)的功能相互作用的潜力,例如TXT的微管蛋白β链基因,CDDP的DNA复制途径,5-FU的DNA合成途径和胸苷酸合成酶基因。
结论:我们新提出了4个分子-AGR2,PDE4D,NINJ2和CDC25B)是预测下咽癌中3种关键化疗药物的细胞反应的强大探索性标志物,也表明它们有可能成为治疗靶标,这可能有助于下咽患者基本化疗的精准医学的发展。(339字)
BACKGROUND: Hypopharyngeal cancer is a relatively rare malignancy with poor prognosis. Current chemotherapeutic algorithm is still far from personalized medicine, and the identification of the truly active therapeutic biomarkers and/or targets is eagerly awaited.
METHODS: Venturing to focus on the conventional key chemotherapeutic drugs, we identified the most correlative genes (and/or proteins) with cellular sensitivity to docetaxel (TXT), cisplatin (CDDP) and 5-fluorouracil (5-FU) in the expression levels, through 3 steps approach: genome-wide screening, confirmation
study on the quantified expression levels, and knock-down and transfection analyses of the candidates. The probable action pathways of selected genes were examined by Ingenuity Pathway Analysis using a large-scale database, The Cancer Genome Atlas.
RESULTS: The first genome-wide screening
study derived 16 highly correlative genes with cellular drug sensitivity in 15 cell lines (|R| > 0.8, P < 0.01 for CDDP and 5-FU; |R| > 0.5, P < 0.05 for TXT). Among 10 genes the observed correlations were confirmed in the quantified gene expression levels, and finally knock-down and transfection analyses provided 4 molecules as the most potent predictive markers-AGR2 (anterior gradient 2 homolog gene), and PDE4D (phosphodiesterase 4D, cAMP-specific gene) for TXT; NINJ2 (nerve Injury-induced protein 2); CDC25B (cell division cycle 25 homolog B gene) for 5-FU- in both gene and protein expression levels. Overexpression of AGR2, PDE4D signified worse response to TXT, and the repressed expression sensitized TXT activity. Contrary to the findings, in the other 2 molecules, NINJ2 and CDC25, there observed opposite relationship to cellular drug response to the relevant drugs. IPA raised the potential that each selected molecule functionally interacts with main action pathway (and/or targets) of the relevant drug such as tubulin β chain genes for TXT, DNA replication pathway for CDDP, and DNA synthesis pathway and thymidylate synthetase gene for 5-FU.
CONCLUSIONS: We newly propose 4 molecules -AGR2, PDE4D,NINJ2 and CDC25B) as the powerful exploratory markers for prediction of cellular response to 3 key chemotherapeutic drugs in hypopharyngeal cancers and also suggest their potentials to be the therapeutic targets, which could contribute to the development of precision medicine of the essential chemotherapy in hypopharyngeal patients. (339 words).