DNA fingerprinting tags novel altered chromosomal regions and identifies the involvement of SOX5 in the progression of prostate cancer
DNA fingerprinting tags novel altered chromosomal regions and identifies the involvement of SOX5 in the progression of prostate cancer
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DOI：
10.1002/ijc.24243
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发表时间：
2009-05-15
影响因子：
6.4
通讯作者：
Chan, Kwok Wah
中科院分区：
文献类型：
--
作者：
Ma, Stephanie;Chan, Yuen Pin;Chan, Kwok Wah
识别与前列腺癌进展相关的基因组改变可能有助于更好地了解这种高度可变的疾病的发展。采用激光捕获显微切割法(LCM)从人前列腺癌根治术标本中获取匹配的正常、癌前高级别前列腺上皮内瘤细胞和浸润性前列腺癌细胞。从这些细胞中，通过一种名为显微解剖档案病变扫描(SMAL)的改进的基于PCR的技术生成比较DNA指纹。重复出现的多态指纹片段被用于标记改变的染色体区域。在细胞带1p31.3、1q44、2p23.1、3p26.3、3q22.3、4q22.3、4q35.2、5q23.2、8q22.3、8q24.13、9q21.3、9q22.32、10q11.21、11p13、12p12.1、13q12.1、16q12.2和18q21.31处有改变区域。提出了周围区域可能含有突变的候选基因，这些突变可能会改变正常的前列腺细胞，使其进入肿瘤阶段。在这些片段中，在所有26个筛选的正常样本中都没有420个碱基的改变，在2个肿瘤中观察到。该片段被克隆、测序并定位于染色体12p12.1。在该区域内，提出了候选基因性别决定区Y-box 5(SOX5)。对SOX5在细胞系、异种移植和人类前列腺标本中的进一步研究，在RNA和蛋白质水平上都发现该基因在肿瘤中过度表达。随后，通过荧光原位杂交发现，这种过度表达是由该区域的扩增引起的。综上所述，我们的结果表明LCM结合SMAI-PCRDNA指纹图谱是一种筛选和鉴定染色体区域和癌症发生相关基因的有用方法。此外，SOX5的过度表达与前列腺癌的进展和远处转移的早期发展有关。(C)2008年Wiley Liss，Inc.
Identification of genomic alterations associated with the progression of prostate cancer may facilitate the better understanding of the development of this highly variable disease. Matched normal, premalignant high-grade prostatic intraepithelial neoplasia and invasive prostate carcinoma cells were procured by laser capture microdissection (LCM) from human radical prostatectomy specimens. From these cells, comparative DNA fingerprints were generated by a modified PCR-based technique called scanning of microdissected archival lesion (SMAL)-PCR. Recurrent polymorphic fingerprint fragments were used in tagging altered chromosomal regions. Altered regions were found at cytobands 1p31.3, 1q44, 2p23.1, 3p26.3, 3q22.3, 4q22.3, 4q35.2, 5q23.2, 8q22.3, 8q24.13, 9q21.3, 9q22.32, 10q11.21, 11p13, 12p12.1, 13q12.1, 16q12.2 and 18q21.31. Candidate genes in the surrounding area that may possibly harbor mutations that change normal prostatic cells to progress into their tumor stages were proposed. Of these fragments, a 420 bp alteration, absent in all 26 normal samples screened, was observed in 2 tumors. This fragment was cloned, sequenced and localized to chromosome 12p12.1. Within this region, candidate gene sex determining region Y-box 5 (SOX5) was proposed. Further studies of SOX5 in cell lines, xenografts and human prostate specimens, at both the RNA and protein levels, found overexpression of the gene in tumors. This overexpression was then subsequently found by fluorescent in situ hybridization to be caused by amplication of the region. In conclusion, our results suggest LCM coupled with SMAI-PCR DNA fingerprinting is a useful method for the screening and identification of chromosomal regions and genes associated with cancer development. Further, overexpression of SOX5 is associated with prostate tumor progression and early development of distant metastasis. (C) 2008 Wiley Liss, Inc.