The Role Of WT1 Gene In Polycystic Ovary Syndrome

Chpter I.The Association Between WT1 and PCOSBACKGROUND:Polycystic ovary syndrome(PCOS)is a heterogeneous disorder characterized by chronic ovulatory dysfunction,hyperandrogenism and polycystic ovaries.Wilms' tumor 1(WT1)encoding a transcription factor involved in the differentiation of granulosa cells(GCs)regulates androgen receptor in the development of male genitalia.However,the expression pattern and possible role of WT1 in ovaries of PCOS patients are still unknown.OBJECTIVE:To detect the association between WT1 and clinical characteristics of PCOS patients and the role of Wtl in the granulosa cells of mice.MATERIALS AND METHODS:GCs from 102 PCOS patients(PCOS group)and 65 healthy controls(control group)were isolated.The expression of WT1 in GCs was quantified using reverse transcription-polymerase chain reaction.The correlation between WT1 expression and clinical characteristics was evaluated in PCOS patients.RESULTS:WT1 expression was increased in PCOS patients compared with the normal controls.The expression of WT1 was moderately correlated with testosterone(r = 0.334,P = 0.001)and luteinizing hormone(r = 0.357,P = 0.001)levels and the antral follicle counts(AFC)(r = 0.337,P= 0.001).Logistic regression analysis showed that WT1 increase the risk in PCOS:OR=1.46,95%CI(1.12?1.92),P=0.006.After adjusting the BMI,the conclusion was not changed,OR=1.388,95%CI(1.07-1.80),Padj=0.014.CONCLUSION:WT1 was overexpressed in granulosa cells of PCOS patients.WT1 is likely to increase the risk in PCOS.Chapter II.The Effect of Tox3 on the Synthesis of Estrogen2 via Wt1BACKGROUND:Through polycystic ovary syndrome related GWAS,Tox3 was identified an susceptible gene to PCOS.TOX3 encodes a calcium dependent transcription factor TOX3.In structure,TOX3 consists of a nuclear location signal at the N terminal,a high mobility group box region and the C terminal.The high migration box region can change the structure of chromosomes in the DNA region.The interaction of calcium binding protein(CBP)complex in rabbit neurons regulates CRE mediated transcription.Our team has found that Wt1 was upregulated after upregulating Tox3 through microarray.Our team has proved that WT1 was expressed highly in the granulosa cells of PCOS patients.Patient granulosa cells are differentiated terminal cells that cannot be passed on and borrowed.OBJECTIVE:To detect if Tox3 affect the expression of Wt1 and the synthesis of estrogen.MATERIALS AND METHODS:We constructed overexpressive vectors of Tox3 and Wtl respectively.Different expression genes were obtained by microarray.We detected the expressive level of Cyp19 and Fshr by Western Blot.We analysed the concentration of E2 in the culture.supernat of cells by Access Immunoassay System.We observe the proliferation and apoptosis of cells by CCK8 and Hochest experiments.RESULTS:WB verified that Wtl was upregulated in granulosa cells with over expression of TOX3.CYP19A1 and FSHR were decreased in Wtl upregulated granulosa cells.What is more,estrogen 2 in culture supernate of Wt1 upregulated granulosa cells was lower than the control.Wtl had no evident effect on the proliferation and apoptosis of granulosa cells.CONCLUSION:This study suggest that Tox3 may involve in inhibiting the differatiation of cells and the product of E2 in granulosa cells through upregulating Wt1,but not through inhibiting the proliferation of granulosa cells.