Functional analysis and application of cancer treatment of unknown gene express in spermatogonial stem cells and cancer cells

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We have isolated PTP-X that is specifically expressed in spermatogonial cells by cDNA RDA subtraction. PTP-X mRNA is expressed at a consistent level during all developmental stage of the testis and testicular tumor cell lines. So, we investigated the regulation of expression and the biological function of PTP-X gene. We compared gene expression between testicular tumors, tumor cell lines and wild type testis by northern blot analysis, but not detected alteration of splicing and expression.We introduced short interference RNA (siRNA) into embryonic stem (ES) cells, and isolated SSEA-1 positive cells from embryoid bodies (EBs). These cells were differentiated to germ cell line in the presence of retinoic acid.We examined the expression of genes implicated in germ cell development.There are no difference expression levels of genes that are expressed in the spermatogonia to spermatocyte. In contrast, expressions of spermatid specific genes were reduced in knock down cells.Next, we examined siRNA chimera testis produced from siRNA ES cells. Expression pattern of germ cell markers was analyzed by immunohistochemistry on sections from frozen siRNA chimera testis or wild testis at 14 weeks. GATA-1 positive cells were presence at seminiferous tubules, but Dmc1 and SCP1 positive cells were presence at lumen.Both in vitro and in vivo results indicate that PTP-X involve in differentiation of spermatogenesis.