Expression profiling in ovarian clear cell carcinoma identification of hepatocyte nuclear factor-1β as a molecular marker and a possible molecular target for therapy of ovarian clear cell carcinoma
Expression profiling in ovarian clear cell carcinoma identification of hepatocyte nuclear factor-1β as a molecular marker and a possible molecular target for therapy of ovarian clear cell carcinoma
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DOI：
10.1016/s0002-9440(10)63605-x
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发表时间：
2003-12-01
影响因子：
6
通讯作者：
Hirohashi, S
中科院分区：
文献类型：
--
作者：
Tsuchiya, A;Sakamoto, M;Hirohashi, S
在所有的上皮性卵巢癌中，卵巢透明细胞癌（CCC）的预后最差。我们应用寡核苷酸芯片技术，以确定基因通常参与CCC。在分析的12,600个相似基因中，28个在4个CCC和7个非CCC细胞系中表达显著不同。在CCC中上调的16个基因中，我们进一步研究了一个转录因子，肝细胞核因子-1 β（hepatocyte nuclear factor-1 beta，HNF-1 beta）。我们使用实时定量逆转录-聚合酶链反应和免疫印迹技术验证了CCC中HNF-1 β在mRNA和蛋白水平上的上调。83例手术切除的卵巢癌的免疫组化分析显示，几乎所有CCC标本（22例中的21例）的细胞核中均有HNF-1 β染色，而大多数非CCC标本（61例中的60例）的细胞核中无免疫染色或仅有灶性和微弱的染色。此外，我们研究了HNF-1 β在CCC中表达的意义，使用RNA干扰。通过RNA干扰减少HNF-1 β表达诱导卵巢CCC细胞凋亡，这一点通过末端dUTP缺口末端标记和荧光激活细胞分选分析得到证实。提示HNF-1 β不仅是一种良好的CCC特异性分子标记物，而且可作为治疗卵巢CCC的分子靶点。
of all of the epithelial ovarian cancers, clear cell carcinoma (CCC) of the ovary has the worst prognosis. We applied the oligonucleotide array technique to identify genes generally involved in CCC. Of the similar to12,600 genes that were analyzed, 28 were expressed significantly differently between four CCC and seven non-CCC cell lines. Among 16 up-regulated genes in CCC, we further investigated a transcription factor, hepatocyte nuclear factor-1beta (HNF-1beta). We validated up-regulation of HNF-1beta in CCC in terms of both mRNA and protein level using real-time quantitative reverse transcriptase-polymerase chain reaction and immunoblotting. Immunohistochemical analysis of 83 surgically resected ovarian cancers showed that almost all CCC specimens (21 of 22 cases) had nuclear staining for HNF-1beta, whereas most non-CCC specimens (60 of 61 cases) showed no immunostaining or only focal and faint staining in the nucleus. Furthermore, we investigated the significance of HNF-1beta expression in CCC using RNA interference. The reduction of HNF-1beta expression by RNA interference induced apoptotic cell death in ovarian CCC cells, which was confirmed by terminal dUTP nick-end labeling and fluorescence-activated cell-sorting analyses. Our results suggest that HNF-1beta is not only an excellent CCC-specific molecular marker but also a molecular target for therapy of ovarian CCC.