Primary Research Of The Clinical Features And Molecular Genetic Mechanism Of A Family With Gonadoblastoma

Gonadoblastoma (GB) is a rare tumor in dysgenetic gonad, the happens of GB is correlated with disorders of sex development. The main cause of GB can be divided into environmental and non-environmental factor, environmental factor included smoking, drinking wine and occupational exposure; and non-environmental factor is the abnormal of genomic DNA in gonadal. Researchers has been identified lots of genes correlated with GB, and Testis specific protein on Y chromosome (TSPY) is regards as the oncogene of GB. Because the development of GB involves multiple genes, the molecular mechanism of the development of GB is still not clear. Here, samples from a family of gonadoblastoma were collected, and the clinical features and genetic characterization of this family were analyzed by different molecular biological technologies, it may be helpful for clarify the mechanism of GB development.Methods:Firstly, the karyotype, biochemical, immunohistochemical and pathological character of this GB family were tested;Secondly, the copy number of 12 candidate genes, WNT4、SRY、ZFY、UTY、 NR5A1、NR0B1、COL4A5、GJB1、PQBP1、CXorf21、PHEX、SOX9 were detected by multiplex ligation-dependent probe amplification (MLPA),Thirdly, the copy number of the genomic genes were analyzed by next generation sequencing,Fourthly, the copy number of TSPY gene is tested by quantificative PCR,The fifth, the SNP and insert and delete mutation of the genomic genes were analyzed by next generation sequencing (deep),The sixth, the exonic and untranscriptic areas of SRY、WT1、DHH、GATA4、 MAP3K1、NR5A1、SOX9、TSPY were tested by Sanger sequencing,Results:1. the karyotype of this GB family is 46, XY, the size of tumor tissue is medium compared with previous reports, and a large parts of tumor is calculium, the expression of OCT3/4 and CD117 proteins were positive by immunohistochemical test,2. Non copy number variations of candidate genes and the genomic genes were found by MLPA and next generation sequencing technology,3. Exon sequencing found six SNPs in rs3829125 (AKR1C4 (exon4:C434G)、 rs4880718 (AKRIC4 (exon7:A749G))、rs17134592 (AKRIC4 (exon9:C931G)、 rs3739583 (DMRTI(exonl:T133A))、rs3824419 (DMRT2 (exon4:G1141C))、 rs11545029 (WWOX (exon5:G196A) which related to DSD happens,4. The missense mutation in TSPY genes (c.615G> T) identified by Sanger sequencing,5. The copy number of TSPY gene is 9-11, less than 30-50 of normal populations,Discussion and conclusion:This report use several molecular biological technologies to study a family with GB, and this research firstly report the TSPY gene exit mutation and the copy number of TSPY gene is low. Based on the comprehensive analysis of the gene interaction network of sex development and sterm cell development, we point out that the lower expression of GB is the primitive cause of GB development, not the higher expression of TSPY protein. Moreover, based on the gene interaction network, we suggested a potential model of GB development which included gene expression level and pathological level. In summary, this research would be helpful to prompt the further research of the mechanism of GB development.