The Role Of PPARG And Its Splice Variant In Polycystic Ovary Syndrome

Objective:(1)To detect the expression levels of PPARG and its splice variant in granulosa cells of polycystic ovary syndrome and control group,and to explore the relationship between PPARG splice variants and clinical phenotype.(2)Stabilize transfected cells by lentivirus to investigate the effects of PPARG splice variant on cell proliferation,migration and apoptosis.(3)To explore the relationship between PPARG splice variant and high androgen,and to detect the effects of PPARG splice variant on the expression of important genes related to metabolism.Design:(1)case-control study;(2)basic researchMaterials and Methods:(1)153 women with polycystic ovary syndrome and 153 normal women who underwent in vitro fertilization-embryo transfer therapy due to tubal factors or male factors were recruited in our hospital.The basal hormone levels in peripheral blood were detected.The granulosa cells were collected after egg retrieval.The full-length sequence of PPARG1 in granulosa cells of PCOS patients was detected by RACE technique.The expression of PPARG and its splice variants in PCOS group and control group were detected by PCR and analyzed with clinical data.(2)The overexpressed cell model stably transfected with PPARG and its deletion splice variant were established using lentivirus.The cells used were identified as ovarian granulosa cell carcinoma cell lines KGN.Then we detected cell proliferation,migration and apoptosis.(3)Using the above cell line,the androgen levels in the cell culture supernatant were measured.Besides,cells were collected after stimulation with a concentration gradient DHT to detect the expression level of PPARG deletion splice variant.The expression levels of important genes in the metabolic pathway after overexpression of PPARG and its deletion splice variant were screened by PCR ARRAY.RESULTS:(1)The expression of PPARG deletion splice variant was detected in both PCOS and control groups.The sequencing results showed that exon5 was deleted.The expression of PPARG was decreased in PCOS group and the expression of its deletion splice variant was increased compared to the control group.(2)Clinical data showed that there were obvious menstrual disorders,higher androgen and estrogen level in PCOS group.Besides,the clinical phenotype of PCOS patients with PPARG deletion splice variant was more obvious.(3)Upregulation of PPARG deletion splice variant inhibits cell proliferation,migration and cell apoptosis.(4)The androgen levels in the supernatant of PPARG deletion splice variant overexpressing cells were significantly higher than those in the control group,while DHT stimulation at different concentrations had no significant effect on the expression level of PPARG deletion splice variant.(5)The high expression of PPARG splice variant mainly caused changes in lipid metabolism related gene expression level.Conclusion: The expression level of PPARG deletion splice variant is increased in granulosa cells of PCOS patients,suggesting that it plays a role in the pathogenesis of PCOS.In addition,high expression of PPARG deletion splice variant inhibited cell proliferation,migration and apoptosis,which may lead to developmental disorders and ovulation disorders of follicular cells,and induced a series of clinical manifestations of PCOS,especially rare ovulation or anovulation.Granulocyte androgen anabolic disorders caused by PPARG deletion splice variant may be one of the mechanisms of PCOS ovary high androgen.Abnormalities in lipid metabolismrelated genes caused by PPARG deletion splice variant may lead to the appearance of a clinically obese phenotype of PCOS.