The Expression And Significance Of FGFR2 In Preputial Tissue With Hypospadias

Hypospadias was anterior urethral hypoplasia, resulting in normal urethral meatus ectopic proximal to the perineum way, some patients with malformation of chordee of penis. It was the one common Congenital malformations in human urogenital system, the incidence in newborns was 250/100,000.Second only to congenital heart disease. The incidence of hypospadias has an upward trend in recent years. Little was known about The etiology of hypspasias was complicated and was not well elucidated。multiple factors were involved in The genesis of hypospadias. It has been demonstrated that the etiology of hypospadias include: gene mutations, endocrine disturbance, intracellular sigal transmation , androgen receptor dysfunction and low expression of epidermal growth factor and environmental factors etc'.Petiot et al.'s reports had discovered that FGFR2-ⅢB absence can provoke hypospadias with animal model.In the FGFR2-ⅢB---/- rat model of hypospadias, urethral signaling regions, as indicated by SHH and FGF8 expression;however,cell proliferation arrests prematurely and maturation of the urethral epithelium is disrupted. FGFR2-ⅢB-/- mutants fail to maintain the progenitor cell population required for uroepithelial renewal during tubular morphogenesis. They show that flutamide antagonism of the androgen receptor (AR) leads to loss of FGFR2-ⅢB and FGF10 expression in the urethra, and an associated hypospadias phenotype, suggesting that these genes are downstream targets of AR during external genital development. In addition, Malformations of the urogenital tract also occur in Beare-Stevenson cutis gyrata syndrome,Split Hand/Split Foot syndrome, limb-mammary syndrome, ectrodactyly-ectodermal dysplasia-cleft lip/palate (EEC) and Hay-Wells syndrome,which was characterized by external genital defects such as hypospadias and bifid scrotum, And Mis-sense mutations in FGFR2 was found in these hereditary disease. Besides that ,by either genetic or environmental factors, may therefore involve negative regulation of the FGFR2 pathway. The studies above indicated that the etiology of hypospadias has intimate corelation with FGFR2 pathway.Currently, environmental factors, genetic mutation, hormone regulation, the abnormal receptor (ER,AR abnormal) most associated with the study of the hypospadias, involving epithelial the urethra, and its development and its associated molecular pathogenesis-related genes and little of this research. The between the development and growth of urethral epithelium and FGFR2 was dosed. But it is not well understood that whether FGFR2 defection lead to human hypospadias and the related molecule molecule mechanism.The expression of FGFR2 protein and mRNA was deteced in the preputial tissue of hypospadias patients and normal control in our exprement to explore the correlation between FGFR2 expression and hypospadias and to laid foundation for futher research of FGFR2 gene function in the pathogenesie of hypospadias.Chapter 1: The expression of tibroblast growth factor receptor 2 in patients' preputial tissue with hypospadias[Objective] To Study the expression of fibroblast growth factor receptor 2 in patients of hypospadias group and the normal control group in preputial tissue for explore the relationships between fibroblast growth factor receptor 2 proteinum and hypospadias.[Methods] Immunohistochemistry and Western blotting technique was used to detect the expression of fibroblast growth factor receptor 2 in both groups.[Results] The expression of fibroblast growth factor receptor 2 in preputial tissue of normal and hypospadias was significantly (P＜0.05,). The expression of fibroblast growth factor receptor 2 in patients with hypospadias was weakened. Between the mild, moderate, severe three groups, the expression of fibroblast growth factor receptor 2 was no statistical significance (P＞0.05) .The FGFR2 protein located in the preputial tissue endochylema of the keratinocyte and fibroblast cell.The amount of protein of Western bloting resulted that the hypospadias patients preputial tissue fibroblast growth factor receptor 2 protein concentrations lower than the normal control group. Correspondence quantitation of FGFR2 protein was 0.39±0.12 in normal control group. Correspondence quantitation of FGFR2 protein was 0.19±0.09 in hypospadias group. Correspondence quantitation of FGFR2 protein was 0.27±0.08 in mild group, 0.16±0.04 in moderate group,0.09±0.03 in severe group . Between Hypospadias group and the normal control group was statistically significant(P＜0.05). In the three groups, no comparison between moderate and severe groups. (P＞0.05).[Conclusion]1, Positive result was coloration in the preputial tissue epidermis fibroblast cell and keratinize cell of basilar part in the preputial tissue.2, Immunohistochemistry : hadro- masculine and masculine in bulk normal preputial tissue, Negative in bulk hypospadias.3,The expression of FGFR2 of preputial tissue in hypospadias was down regulation than normal.The degression in hypospadias patients's preputial tissue of the expression of fibroblast growth factor receptor 2 suggested that the relationship of fibroblast growth factor receptor 2 and hypospadias was closely. Further research of FGFR2 gene function in hypospadias may reveal the pathogenesis of hypospadias.Chapter 2 The expression of transcriptional level of Fibroblast growth factor receptor 2 in preputial tissue with hypospadias[Objective] To detect the expression of fibroblast growth factor receptor 2 mRNA in hypospadias patients's preputial tissue. Exploration the relationships of hypospadias and fibroblast growth factor receptor 2 gene function.[Methods] We used RT-PCR and Real time PCR technique to detect the expression of fibroblast growth factor receptor 2 mRNA in both groups.[Results] The expression of hypospadias patients's fibroblast growth factor receptor 2 mRNA was1.22±0.052,the normal wasl.30±0.069. Between the two groups was significant(P＜0.05). Mild and moderate of hypospadias in those groups ,the expression of fibroblast growth factor receptor 2 mRNA was no significant(P＞0.05). Based on the statistics, The expression of fibroblast growth factor receptor 2 mRNA in severe levels lower than the other two groups.[Conclusion] The degression of fibroblast growth factor receptor 2 in hypospadias patients.It cued that the relationship of fibroblast growth factor receptor 2 gene and hypospadias was closely.The further study of the fibroblast growth factor receptor 2 gene maybe explore the pathogenesy of hypospadias.