Cancer Genomics Program, Peter MacCallum Cancer Centre, East Melbourne, Australia.
PLoS One. 2010 Nov 12;5(11):e15498. doi: 10.1371/journal.pone.0015498.
Genomic amplification of 19q12 occurs in several cancer types including ovarian cancer where it is associated with primary treatment failure. We systematically attenuated expression of genes within the minimally defined 19q12 region in ovarian cell lines using short-interfering RNAs (siRNA) to identify driver oncogene(s) within the amplicon. Knockdown of CCNE1 resulted in G1/S phase arrest, reduced cell viability and apoptosis only in amplification-carrying cells. Although CCNE1 knockdown increased cisplatin resistance in short-term assays, clonogenic survival was inhibited after treatment. Gain of 20q11 was highly correlated with 19q12 amplification and spanned a 2.5 Mb region including TPX2, a centromeric protein required for mitotic spindle function. Expression of TPX2 was highly correlated with gene amplification and with CCNE1 expression in primary tumors. siRNA inhibition of TPX2 reduced cell viability but this effect was not amplicon-dependent. These findings demonstrate that CCNE1 is a key driver in the 19q12 amplicon required for survival and clonogenicity in cells with locus amplification. Co-amplification at 19q12 and 20q11 implies the presence of a cooperative mutational network. These observations have implications for the application of targeted therapies in CCNE1 dependent ovarian cancers.
19q12 基因扩增发生在多种癌症类型中,包括卵巢癌,其中它与原发性治疗失败有关。我们使用小干扰 RNA(siRNA)系统地减弱卵巢细胞系中最小定义的 19q12 区域内的基因表达,以鉴定扩增子内的驱动癌基因。CCNE1 的敲低导致 G1/S 期停滞,仅在携带扩增的细胞中降低细胞活力和凋亡。尽管 CCNE1 敲低在短期测定中增加了顺铂耐药性,但克隆存活在治疗后受到抑制。20q11 的增益与 19q12 扩增高度相关,跨越包括 TPX2 在内的 2.5 Mb 区域,TPX2 是有丝分裂纺锤体功能所必需的着丝粒蛋白。TPX2 的表达与原发性肿瘤中的基因扩增和 CCNE1 表达高度相关。TPX2 的 siRNA 抑制降低了细胞活力,但这种效应与扩增子无关。这些发现表明,CCNE1 是 19q12 扩增子中存活和克隆性所必需的关键驱动基因。19q12 和 20q11 的共扩增意味着存在协同突变网络。这些观察结果对 CCNE1 依赖性卵巢癌中靶向治疗的应用具有重要意义。