| Wilms tumor is one of the most common solid tumors in children. The Wilms' tumor 1 gene ( WT -1) has been identified as a tumor suppressor gene involved in the etiology of Wilms' tunior. Approximately 5-25% of all Wilms' tumors carry mutations in the WT, gene. Alterations in the WTj gene have also been observed in other tumor types, such as leukemia, mesothelioma and desmoplastic small round cell tumor. Dependent on the tumor type, WT - 1 proteins might either function as tumor suppressor proteins or as survival factors. The Wilms tumor suppressor gene WT, encodes a zinc finger protein, expressed as different splicing variants, that has all the hallmarks of a transcription factor. The - KTS form of WT, displays a homogeneous localization within the nucleus and has been shown to activate or repress the activity of various target genes. In contrast, the WT, ( + KTS) variant demonstrates a speckled pattern of expression within the nucleus. This and its association with factors of the splicing machinery has led to the hypothesis that WTt ( + KTS) might play a role in post -transcriptional processes.Mutations in the WT -1 gene can also result in congenital abnormalities as observed in Denys - Drash and Frasier syndrome patients. Mouse models have proven the critical importance of WT - 1 expression for the development of several organs, including the kidneys, the gonads and the spleen. The WT -1 proteins seem to perform two main functions. They regulate the transcription of a variety of target genes and may be involved in post - transcriptional processing of RNA. These isoforms have partially distinct biological functions and effects, which in many cases are also specific for the model system in which WT - 1 is studied. This discusses the molecular mechanisms by which the various WT, isoforms exert their functions in normal development and how alterations in WT - 1 may lead to developmental abnormalities and tumor growth. , Nephroblastomas ( Wilms' tumors) are curable with survival rates about 80%. Wilms' yumor pa-tients can now be achieved using combination therapy with chemotherapy, surgery, and in some cases radiotherapy. But these methods can produce more damages to children than to adults. So we hope a new approaches to therapy Wilms' tumor patients. In addition to there is little research about the biology of nephroblastoma in our country, so many work need to do.PURPOSES( 1) Our purposes is to study the expression of both genes WT - 1 and EGR - I in Wilms' tumor using an immunohistochemical approach. We analysis the expression of both genes in normal kidney tissues , in the blastema! , epithelial component of Wilms' tumor and stromal tissue . We hope to detect the association between the expression of WT - 1 and EGR -1 in the three cells of Wilms' tumor and the clinical progression. morever,We study the relationship between WT - 1 and EGR -1 expression in clinical nephroblastomas. (2) We want to get 32P labled cDNA probe by Reverse Transcription - PCR , and then find WT - 1 gene through in situ blot with the the colon of cDNA libarary of embryomic kidney tissue. (3) To determine whether Wilms' tumor cell lines are suppressed by WT - Igene, we performed chromosome transfer experiments : transfect WT -1 gene into the pediatric kidney - derived cell line G401. we want to Detecting the effects on the population doubling times of the cell line, proliferative capacity , the expression of WT - 1 gene in G401 cell line,and the apoptosis of G401.METHODS(1) Immunohistochemistry: The following primary antibodies were used: a mouse monoclonal antibody against WT - 1; and rabbit polyclonal antibody a-gainst, The peroxidase - antiperoxidase technique was used. Serial sections from all samples which were incubated overnight, in a 60 incubator. After dewa-xing in fresh xylene for 10 min and rehydration in 100% methanol for 10 min, the sections were rinsed in methanol containing 3% hydrogen peroxide for 20min to block endogenous peroxidase activity. Slides were incubated in PBS ?5% BSA for 15 min a...
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