Genetic analysis in human prostate carcinoma detected by two-dimensinal gel electrophoresis (RLGS method)

为了探索人前列腺癌的分子异常，从3种前列腺细胞系LNCaP、PC-3和DU-145中提取的基因组DNA，使用限制性标志基因组扫描（RLGS）方法进行检测，该方法是一种二维凝胶分析，可以评估大约2，000个Not I标志。在LNCaP、PC-3和DU-145中分别检测到24、18和23个扩增位点。11个斑点在所有3个细胞系中普遍增强，扩增范围为正常的2.1至134.1倍。对6例异质性前列腺癌的基因组DNA以及肿瘤内单个和组织学上不同的病灶的基因组DNA进行了检测。在这项研究中，癌症DNA与正常前列腺DNA对照的比较产生了至少35个点的改变。尽管肿瘤的组织学分级存在差异，但与对照相比，所有肿瘤样品共有的3个斑点显示出强度的一致扩增，其他8个共同斑点显示出强度的一致降低。此外，从单个肿瘤内分离的组织学相同的病灶之间发生斑点改变。在RLGS-DNA图谱中检测到的这些斑点变化反映了尚未鉴定的癌基因和抑癌基因的畸变，也表明前列腺癌不仅在组织学上是异质性和多灶性的，而且在遗传学上是多中心的。相反，在检查的16个增生中，每个增生中发现2 - 10个斑点的强度改变，与来自正常前列腺的等同斑点相比，但是对于任何增生样品，可以检测到很少的共同斑点分布。在某些形式的人类良性前列腺增生的发病机制中可能没有一致的遗传变化。这些结果表明，常见的遗传异常可能发生在人类前列腺癌。

To explore the molecular abnormalities in human prostate carcinome, the genomic DNAs extracted from 3 prostate cell lines-LNCaP,PC-3 and DU-145-were examined using restriction landmark genomic scanning (RLGS) methodology, a 2-dimensional gel analysis which allows evaluation of approximately, 2,000 Not I landmarks. The 24,18 and 23 amplified spots were detected in LNCaP,PC-3 and DU-145 DNA,respectively. Eleven spots were commonly intensified in all 3 cell lines, with a range of amplification of 2.1 to 134.1-fold over normal. Alterations in the genomic DNAs of 6 heterogeneous prostate carcinomas, as well as that of indiviual and histologically distinct foci within the tumors, were examined. In this study, comparison of cancer DNAs against normal prostate DNA controls yielded alterations in at least 35 spots. Despite differences in the histological grading of tumors, 3 spots common to all tumor samples showed consistent amplification of intensity and 8 other common spots demonstrated consistent reduction of intensity when compared to control. In addition, spot alterations occurred between histologically identical foci isolated from within single tumors. It is suggested that these spot changes detected in RLGS-generated DNA profiles reflect aberrations in as yet unidentified oncogenes and tumor suppressor genes, and indicate, as well, that prostate cancer is not only histologically heterogeneous and multifocal but also genetically multicentric.On the contrary, within each of the 16 hyperplasias examined, 2 to 10 spots were found with altered intensities, as compared to equivalent spots from normal prostate, but could detect few common spot profiles for any hyperplasia samples. There may be no consistent genetic changes in the pathogenesis of some forms of human benign prostatic hyperplasia. These results suggest that common genetic abnormalities may occur in carcinomas of the human prostate.