Franke F E, Pauls K, Kerkman L, Steger K, Fink L, Burkhardt E, Klonisch T, Bergmann M, Danilov S M
Institut für Pathologie, Justus-Liebig-Universität Giessen.
Verh Dtsch Ges Pathol. 2000;84:199-209.
CD143 (angiotensin I-converting enzyme) occurs in two isoforms: a testicular form (tCD143) expressed during spermatogenesis, and a somatic form (sCD143) generally found in certain other cell types. To study these isoforms in normal and neoplastic germ cells of humans, we analyzed a broad collective of different testicular germ cell tumors (GCTs) of adults, adjacent intratubular germ cell neoplasms (IGCNs), and testicular tissues representing the regular germ cell development. Different techniques were employed on fresh frozen and formalin-fixed, paraffin-embedded tissues: CD143-mRNAs were analyzed by RT-PCR on selected cells after UV-laser-assisted cell picking and by in-situ hybridization using cRNA probes; the proteins were analyzed by semi-quantitative immunohistochemistry using monoclonal antibodies to CD143, and to PLAP/GCAP as controls. In contrast to normal germ cells bearing only tCD143 during spermiogenesis, both mRNA and protein of sCD143 were detected in neoplastic cells of all IGCNs and in the majority of seminomas. sCD143 expression also was found during testicular development, but was differently regulated in fetal germ cells and in GCTs compared with PLAP/GCAP. Thus, our findings (i) demonstrate profound changes in the expression of both CD143 isoforms during regular germ cell development and maturation, (ii) suspect sCD143 being involved in the regulation of germinal stem cell proliferation, (iii) are in agreement with the concept of an 'embryonic state' of neoplastic germ cells, (iv) indicate a close molecular relationship between IGCN and seminoma and, finally, (v) suggest sCD143 as an appropriate marker in the diagnosis of seminomas in addition to PIAP/GCAP.
CD143(血管紧张素I转换酶)存在两种同工型:一种是在精子发生过程中表达的睾丸型(tCD143),另一种是通常在某些其他细胞类型中发现的体细胞型(sCD143)。为了研究人类正常和肿瘤性生殖细胞中的这些同工型,我们分析了大量成年男性不同类型的睾丸生殖细胞肿瘤(GCT)、相邻的管内生殖细胞肿瘤(IGCN)以及代表正常生殖细胞发育的睾丸组织。对新鲜冷冻组织和福尔马林固定、石蜡包埋组织采用了不同技术:通过紫外线激光辅助细胞挑选后对选定细胞进行RT-PCR分析CD143-mRNA,并使用cRNA探针进行原位杂交;使用针对CD143的单克隆抗体通过半定量免疫组织化学分析蛋白质,并以PLAP/GCAP作为对照。与精子形成过程中仅携带tCD143的正常生殖细胞不同,在所有IGCN的肿瘤细胞以及大多数精原细胞瘤中均检测到了sCD143的mRNA和蛋白质。在睾丸发育过程中也发现了sCD143的表达,但与PLAP/GCAP相比,其在胎儿生殖细胞和GCT中的调控方式不同。因此,我们的研究结果(i)表明在正常生殖细胞发育和成熟过程中,两种CD143同工型的表达发生了深刻变化;(ii)怀疑sCD143参与了生殖干细胞增殖的调控;(iii)与肿瘤性生殖细胞“胚胎状态”的概念一致;(iv)表明IGCN和精原细胞瘤之间存在密切的分子关系;最后,(v)提示除PIAP/GCAP外,sCD143可作为精原细胞瘤诊断的合适标志物。