Mutation Study Of Human GDF-9 And BMP-15 Genes In Chinese Han Patients With Polycystic Ovary Syndrome

PartⅠMutation Analysis of the Growth Differentiation Facor-9 Gene in Chinese Han Patients with Polycystic Ovary SyndromeObjectives Polycystic ovary syndrome (PCOS) is one of the common defects which cause ovary dysfunction and link to the aberrant process of folliculogenesis.The preantral follicles growed excessively but the dominant follicle growed blocked are its pathophysiology characteristic.Growth differentiation factor-9 (GDF-9) is originatesd by ovocyte, belongs to the transformation growth factor-Beta (TGF-β) family.Recently study have discovered that in the PCOS patients GDF-9 expressed delayed and lower. In this current study we focused on the mutational analysis of the coding region of GDF-9 gene to find its relations with PCOS.Methods 1.Collected pcos and normal comparison blood preparation each 200 examples, record each name, age, first menstruation age, marriageable age, menstrual cycle, sterile history, height, body weight and so on, measured the foundation sex hormone: estradiol (E2), follicle stimulating hormone(FSH), luteinzing hormone (LH)), testosterone (T), prolactin(PRL). 2.With the blood genome team DNA extraction kit, extract DNA according to the explanation from 500ul peripheral blood. 3.Used primer premeir5 software design primers, by Subsequent polymerase chain reaction (PCR) to amplify the goal fragment. 4.Used DNA automatic sequencer (model: ABI3730XL) bidirectional sequencing all the PCR products .Result(s) Read out the sequencing maps by the chromas software, and compared in GENEBANK. In the coding region of GDF-9 gene, totally ten variants were found. Seven missense mutations (p.Asn5Leu, p.Leu40Val, p.Met45Val, p.Tyr342Phe, p.Ser425Arg, p.R146C, p.Ser428Thr) presented in different patients discretely, and three synonymous mutations (c.169G>T,c.447C>T and c.546G>A) discovered in both patients and controls. Five from the seven missense mutations were firstly discovered in Chinese pcos patients, namely c.15C>G, c.118T>G, c.133A>G, c.1025A>T, c.1275C>A. The rest two mutations(c.436C>T,c.1283G>C) have been reported before.Conclusion(s) The missense mutations in GDF-9 gene indicate this gene is potentially associated with PCOS in Chinese Han women.PartⅡIdentification of 4 Novel Missense Mutations of Human BMP15 in Chinese Han Women with Polycystic Ovary SyndromeObjectives To investigate whether bone morphogenetic protein-15 (BMP-15)mutation is present in Chinese Han women with polycystic ovary syndrome (PCOS).Methods Subsequent polymerase chain reaction (PCR) was employed to extend the candidated gene for the blood of 200 women with pcos, and 200 normal women. The amplified DNA was analyzed by direct sequencing. Results Of six missense mutations were found in the coding region of the BMP-15 gene in pcos patients, four novel missense mutations in BMP-15 were discovered, namely c.34C>G, c.109G>C, c.169C>G, c.288G>C,and two mutations(c.308A>G,c.788insTCT) have been reported.There is one snp( c. 598C>T )discovered in both patients and controls.Conclusions The missense mutations in the BMP-15 gene indicate this gene is potentially associated with PCOS in Chinese Han women.