Differential gene expression profiles between tumor biopsies and short-term primary cultures of ovarian serous carcinomas: Identification of novel molecular biomarkers for early diagnosis and therapy
Differential gene expression profiles between tumor biopsies and short-term primary cultures of ovarian serous carcinomas: Identification of novel molecular biomarkers for early diagnosis and therapy
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DOI：
10.1016/j.ygyno.2006.03.056
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发表时间：
2006-11-01
影响因子：
4.7
通讯作者：
Santin, Alessandro D.
中科院分区：
文献类型：
--
作者：
Bignotti, Eliana;Tassi, Renata A.;Santin, Alessandro D.
目的：通过基因表达谱分析确定对卵巢癌早期诊断和治疗有用的新型分子生物标志物。比较速冻卵巢浆液性癌与匹配的高纯度原代肿瘤细胞培养物的基因指纹。
方法：使用与超过14500个人类基因互补的寡核苷酸微阵列，分析19例速冻卵巢浆液性乳头状癌（OSPC）的基因表达谱，并与15例对照（高纯度人卵巢表面上皮短期培养物，HOSE）进行比较。此外，将5例体外培养不到2周的高纯度原代OSPC的基因表达谱与从匹配样本中获取的速冻卵巢癌活检组织进行比较。采用定量逆转录聚合酶链反应（RT - PCR）和免疫组化（IHC）染色技术，在RNA和蛋白质水平上对一些差异表达基因的微阵列数据进行验证。
结果：对基因表达数据的无监督分析很容易将正常组织与速冻OSPC区分开来，并确定了在OSPC中与HOSE相比分别表现出>3倍上调或下调的901个和557个基因。乳腺珠蛋白2，一种卵巢分泌蛋白，被确定为OSPC中差异表达最显著的基因（19例OSPC中有19例，而15例HOSE中为0例），相对于HOSE上调超过827倍。与HOSE相比，在OSPC中发现紧密连接蛋白和激肽释放酶家族蛋白，包括产气荚膜梭菌肠毒素受体紧密连接蛋白3和4、激肽释放酶6、7、8、10、11以及免疫调节分子B7 - H4是高度过表达的基因之一。发现速冻OSPC的基因指纹与纯化的原代OSPC短期体外培养物的基因指纹高度相关，两组之间仅有8637个基因中的31个（0.35%）差异表达。
结论：原代卵巢癌的短期体外培养可大大提高用于基因表达谱分析的卵巢肿瘤RNA的纯度，而不会导致OSPC指纹发生重大改变。乳腺珠蛋白2、激肽释放酶6、7、8、10、11、紧密连接蛋白3和4以及B7 - H4基因表达产物是具有很大潜力的候选生物标志物，可用于OSPC患者的早期筛查和治疗。（c）2006爱思唯尔公司。保留所有权利。
Objective. To identify novel molecular biomarkers useful for the early diagnosis and therapy of ovarian cancer by gene expression profiling. To compare the genetic fingerprints of flash-frozen ovarian serous carcinomas to those of matched highly purified primary tumor cell cultures.Methods. Gene expression profiles of 19 flash-frozen ovarian serous papillary carcinoma (OSPC) were analyzed and compared to 15 controls (highly purified human ovarian surface epithelium short-term cultures, HOSE) using oligonucleotide microarrays complementary to > 14,500 human genes. In addition, gene expression profiling of 5 highly purified primary OSPC cultured in vitro for less than 2 weeks was compared to flash-frozen ovarian carcinoma biopsies obtained from matched samples. Quantitative RT-PCR and IHC staining techniques were used to validate microarray data at RNA and protein levels for some of the differentially expressed genes.Results. Unsupervised analysis of gene expression data readily distinguished normal tissue from flash-frozen OSPC and identified 901 and 557 genes that exhibited > 3-fold up-regulation or down-regulation, respectively, in OSPC when compared to HOSE. Mammaglobin 2, an ovarian secreted protein, was identified as the top differentially expressed gene in OSPC (19 out 19 OSPC versus 0 out of 15 HOSE) with over 827-fold up-regulation relative to HOSE. The claudin and kallikrein family of proteins including the clostridium perfringens enterotoxin receptors claudin 3 and 4, kallikreins 6, 7, 8, 10, 11 and the immunomodulatory molecule B7-H4 were found among the most highly overexpressed genes in OSPC when compared to HOSE. Genetic fingerprints of flash-frozen OSPC were found to have high correlation with those of purified primary OSPC short-term in vitro cultures with only 31 out of 8637 genes (0.35%) differentially expressed between the two groups.Conclusions. Short-term in vitro culture of primary ovarian carcinomas may greatly increase the purity of ovarian tumor RNA available for gene expression profiling without causing major alteration in OSPC fingerprints. Mammaglobin 2, kallikreins 6, 7, 8, 10, 11, claudin 3 and 4 and B7-H4 gene expression products represent candidate biomarkers endowed with great potential for early screening and therapy of OSPC patients. (c) 2006 Elsevier Inc. All rights reserved.