Ottesen Anne Marie, Skakkebaek Niels E, Lundsteen Claes, Leffers Henrik, Larsen Jacob, Rajpert-De Meyts Ewa
Department of Growth and Reproduction, Juliane Marie Centre, University Hospital of Copenhagen, Copenhagen, Denmark.
Genes Chromosomes Cancer. 2003 Oct;38(2):117-25. doi: 10.1002/gcc.10244.
High-resolution comparative genomic hybridization (HR-CGH) analysis was performed on DNA purified from laser-capture microdissected carcinoma in situ (CIS) cells from nine cases of CIS, either from tissue without any invasive tumor or from testicular parenchyma adjacent to seminoma, nonseminoma, or a combined germ cell tumor. Before CGH analysis, DNA was amplified by degenerate oligonucleotide primed PCR (DOP-PCR) and directly labeled with a mixture of FITC-dUTP and FITC-dCTP. CGH analysis revealed extra chromosome arm 12p material in six out of seven cases with CIS adjacent to overt tumors, but only a diminutive gain of 12q was noted in one of the two cases of CIS without invasive elements. In addition, gains of parts of chromosome 8 (3/7) and losses of chromosome 5 (2/7) were demonstrated in CIS adjacent to invasive tumors. Gains of parts of chromosome 7 were found in CIS adjacent to seminoma (4/4), whereas relative gains of chromosome 15 were identified in some cases of CIS adjacent to seminoma and in isolated CIS in comparison to CIS adjacent to nonseminoma. Our data seem to indicate that extra 12p material is not present in the "dormant" CIS cell before development of an invasive tumor. The gain of extra chromosome 12 material may not be an early event in the neoplastic transformation, but is most likely associated with a more malignant progression of the CIS cell.
对从9例原位癌(CIS)的激光捕获显微切割细胞中纯化的DNA进行了高分辨率比较基因组杂交(HR-CGH)分析,这些原位癌病例来自无任何浸润性肿瘤的组织,或来自与精原细胞瘤、非精原细胞瘤或混合性生殖细胞肿瘤相邻的睾丸实质。在进行CGH分析之前,通过简并寡核苷酸引物PCR(DOP-PCR)扩增DNA,并直接用FITC-dUTP和FITC-dCTP混合物进行标记。CGH分析显示,在7例与明显肿瘤相邻的CIS病例中,有6例存在额外的12号染色体短臂物质,但在2例无浸润成分的CIS病例中,仅1例发现12号染色体长臂有微小增加。此外,在与浸润性肿瘤相邻的CIS中,还发现了8号染色体部分区域的增加(3/7)和5号染色体的缺失(2/7)。在与精原细胞瘤相邻的CIS中发现了7号染色体部分区域的增加(4/4),而与非精原细胞瘤相邻的CIS相比,在一些与精原细胞瘤相邻的CIS病例和孤立的CIS中发现了15号染色体的相对增加。我们的数据似乎表明,在浸润性肿瘤发生之前,“休眠”的CIS细胞中不存在额外的12号染色体短臂物质。额外的12号染色体物质的增加可能不是肿瘤转化的早期事件,而很可能与CIS细胞更恶性的进展有关。