Tsunoda Toshiyuki, Furusato Bungo, Takashima Yasuo, Ravulapalli Suma, Dobi Albert, Srivastava Shiv, McLeod David G, Sesterhenn Isabell A, Ornstein David K, Shirasawa Senji
Faculty of Medicine, Department of Cell Biology, Fukuoka University, Nanakuma, Jonan-ku, Fukuoka, Japan.
Prostate. 2009 Sep 15;69(13):1398-403. doi: 10.1002/pros.20988.
Three-dimensional culture (3DC) is a relevant in vitro model used to study prostate development and carcinogenesis. Recent studies have indicated that 3DC-associated genes would be more sensitive as prognostic markers for cancer; however, no 3DC-associated genes in prostate cancer (CaP) have thus far been elucidated.
Candidate 3DC-associated genes in non-malignant prostatic epithelial cells, RWPE-1 and TA2, were selected, based on a comparison of microarray gene expression data between cells grown in two-dimensional culture (2DC) and in 3DC. To extract CaP-associated genes among the 3DC-associated genes, gene expression levels from the microdissected tissue samples were compared between 20 well-differentiated or 20 poorly differentiated CaP, as well as matched normal prostate epithelium. The expression levels of CaP-associated genes in 3DC were validated by quantitative RT-PCR using TA2 cells in 2DC and 3DC. Protein expression of periostin (POSTN), which is one of the 3DC CaP-associated genes, was further evaluated in the clinical samples by immunohistochemistry.
Several genes including POSTN were identified as CaP-associated genes using a 3DC system. Immunohistochemical analyses revealed that POSTN expression was increased in the early stages of CaP (Gleason score 6-7), but not in the advanced stages of CaP. Furthermore, the positive ratio observed for the expression of POSTN in tumor-associated stroma was significantly correlated with the degree of malignancy.
POSTN, one of the 3DC-associated genes, may serve as a potential biomarker for predicting the pathological grade and prognosis of CaP. Prostate 69: 1398-1403, 2009. (c) 2009 Wiley-Liss, Inc.
三维培养(3DC)是用于研究前列腺发育和致癌作用的一种相关体外模型。近期研究表明,与3DC相关的基因作为癌症预后标志物可能更具敏感性;然而,迄今为止,前列腺癌(CaP)中尚未阐明任何与3DC相关的基因。
基于二维培养(2DC)和3DC中生长的细胞之间的微阵列基因表达数据比较,选择非恶性前列腺上皮细胞RWPE - 1和TA2中与3DC相关的候选基因。为了在与3DC相关的基因中提取与CaP相关的基因,比较了20例高分化或20例低分化CaP以及匹配的正常前列腺上皮组织微切割样本中的基因表达水平。使用TA2细胞在2DC和3DC中通过定量RT - PCR验证了3DC中与CaP相关基因的表达水平。通过免疫组织化学进一步评估了临床样本中骨膜蛋白(POSTN)的蛋白表达,POSTN是与3DC CaP相关的基因之一。
使用3DC系统鉴定了包括POSTN在内的几个基因作为与CaP相关的基因。免疫组织化学分析显示,POSTN表达在CaP早期(Gleason评分6 - 7)增加,但在CaP晚期未增加。此外,在肿瘤相关基质中观察到的POSTN表达阳性率与恶性程度显著相关。
POSTN作为与3DC相关的基因之一,可能作为预测CaP病理分级和预后的潜在生物标志物。《前列腺》69: 1398 - 1403, 2009。(c)2009威利 - 利斯公司。