The susceptibility of primordial germ cells to malignant transformation and isolation and characterization of members of a new gene family differentially expressed in invasive and non-invasive immortalized male germ cells

摘要: In this study, we examined the susceptibility of primordial germ cells (PGCs) to malignant transformation using a transgenic mouse model. model used simian virus 40 (SV40) large tumor antigen (LTA) fused promoter cell alkaline phosphatase (GCAP) specifically direct oncogenic activity SV40-LTA PGCs. Human GCAP is developmentally expressed in and also from 2-cell blastocyst stage preimplantation development reappears trace amounts adult testis. Furthermore, reexpressed precancerous stages testicular tumors (carcinoma situ) more differentiated tumors. We analyzed regulatory sequences 5´-flanking region human gene which are responsible for its expression different determine minimal sufficient high tissue specific GCAP. Three independent lines were generated fragment 1.7 kb 5´flanking as promoter. RT-PCR analysis showed testis LTA adults heterozygous homozygous mice amounts. could detect mRNA 8.5 dpc prenatal embryos. However, even after period about 20 months, no was observed any organ these mice. Using an "in vitro" approach analysing differentially genes GC-1spg through suppression subtractive hybridization (SSH), so far unknown found be highly invasive identified, named Pfcp (PHD-finger-like chromatin proteins) deduced protein sequence shows domain known "PHD-finger" like domain, involved binding. The cDNA isolated well genomic structure. pattern cellular subcellular location determined: screening cosmid library Southern blot family consists four murine genes. Northern one transcript at approximately 1.2 kb. tissues ubiquitous. PFCP ubiquitous well. localized nucleus binds structures interphase metaphase cycle.