| Objective: To detect the association between FTO gene variants(rs79206939A/G,rs9930506A/G,rs8050136A/C and rs1588413C/T)and PCOS susceptibility,as well as clinical biochemical data and IVF outcomes.We also detect the expression of FTO,IRX3 and IRX5 proteins in different phenotypes.Methods: A total of 516 subjects including 333 PCOS patients and 183 healthy females were recruited from reproductive health hospital of Nanhua-Xinghui from December 2015 to December 2017.PCOS was defined according to the criteria of Rotterdam Revised 2003,and the controls was defined when the infertility caused by the male.A total of 2m L peripheral blood was collected,1 m L was used to extract DNA,the rest of sample was used to define some clinical biochemical data.On-off switch and a classic polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)were used to genotyping.We divided the whole sample into 4 groups: PCOS and obesity,PCOS and non-obesity,control and obesity,control and non-obesity.The protein of FTO,IRX3 and IRX5 were extracted respectively from follicular fluid and granule cells.The expression the proteins were analyzed by Western-Blot.IVF outcomes were assessed the relationship to the FTO gene variants too.Results:1.For rs79206939A/G,the frequencies of AA,AG and GG in PCOS group were respectively 3.0%,23.1%,73.9%,and 3.3%,16.4%,80.3% in controls(p=0.197),so there was no statistic difference.In the recessive model,the frequencies of AA+AG and GG in PCOS group were respectively 26.1%,3.9% and 19.7%,80.3% in controls(p=0.100),so there was no statistic difference.Allele frequencies of A and G in PCOS group were respectively 14.6%,85.4%,and 11.5%,88.5% in controls(p=0.164),no statistic difference was observed.No significant differences were detected in the clinical biochemical data in both PCOS and controls.Compared with GG subgroup,the high qualified embryo rate and implantation rate in AA+AG subgroup were higher in the IVF outcomes of PCOS(p=0.020;0.018).2.For rs9930506A/G,the GG,AG and AA frequencies in PCOS were respectively 1.2%,30.9%,67.9%,and 1.1%,27.3%,71.6% in controls(p=0.682),no statistic difference was detected.There is no statistic difference in the recessive model,as the frequencies of GG+AG and AA were respectively 31.2%,68.8%,and 28.4%,71.6% in controls(p=0.382).No significant difference was also found in allele frequencies.Compared with AA subgroup,the level of testosterone(T)in the GG+AG subgroup was significantly lower(p=0.008).No significant differences were detected between GG+AG and AA subgroup in the IVF outcomes of PCOS.3.For rs8050136A/C,the frequencies of AA,AC and CC in PCOS were respectively 1.5%,15.3%,83.2%,followed 1.0%,7.1%,91.9% in controls(p=0.009),so there is significant difference in additive model.The frequencies of AA+AC and CC were respectively 16.8%,83.2%,and 8.2%,91.8% in controls(p=0.007).Significant difference was also discovered in allele frequencies,as the frequencies of A and C in PCOS were respectively 9.2%,90.8%,followed 4.6%,95.4% in controls(p=0.023).Compared with CC subgroup,the level of follicle-stimulating hormone(FSH)in AA+AC was lower(p=0.006),but the level of body mass index(BMI)was higher(p=0.030).And ovulation number in AA+AC was higher than CC in the IVF analysis of PCOS(p=0.015).4.For rs1588413C/T,the frequencies of TT,CT and CC in PCOS were respectively 1.2%,46.8%,52.0%,followed 1.6%,35.5%,62.9% in controls(p=0.045).Significant difference was discovered in the recessive model,as the frequencies of TT+CT and CC were 48.0%,52.0%,and 37.2%,62.8% in controls(p=0.017).But interestingly,no significant difference was no defined in allele frequencies(p=0.055).Compared with CC subgroup,those with TT+CT genotype had higher luteinizing hormone(LH)and estradiol(E2)level(p=0.018;0.041 respectively).Also,patients with CC genotype had higher implantation rate than those with TT+CT subgroup(p=0.024).5.In follicular fluid,FTO protein expression in obesity was higher than non-obesity.However,the expression of IRX3 was lower in obesity compared with non-obesity group.IRX3 protein expression is also different between PCOS&obesity and control&obesity groups.The expression of IRX5 in PCOS&obesity was higher than in PCOS&non-obesity and control&obesity,and it in control&obesity was higher than in PCOS&obesity and control&non-obesity.6.In granule cells,FTO protein expression in obesity was higher than non-obesity group just like in follicular fluid.And FTO protein expression was higher in PCOS&obesity than in control&obesity group.As for IRX3,the expression of it in obesity or PCOS was lower than in non-obesity or control groups.IRX5 protein expression in PCOS was higher than controls,and in PCOS group,it was higher in PCOS&obesity subgroup compared with PCOS&non-obesity subgroup.Conclusions:1.rs8050136A/C and rs1588413C/T variants may be associated with PCOS susceptibility.2.rs9930506A/G variant is associated with the level of T in PCOS;rs8050136A/C variant is associated with the levels of BMI and FSH in PCOS;and rs1588413C/T SNP has a relationship with the levels of LH and E2 in PCOS.3.FTO expression in obesity is significantly higher than in non-obesity group no matter what in follicular fluid or granule cells,while the IRX3 expression was opposite.IRX3 expression in obesity is lower than in non-obesity group.There are many factors influence the expression of IRX5,such as PCOS or not,obesity or not.And it is different for IRX5 expression trend in follicular fluid and granule cells.4.rs79206939A/G variant is associated with high qualified embryo rate and implantation rate in PCOS.rs8050136A/C variant is associated with ovulation number in PCOS.rs1588413C/T variant has a relationship to implantation rate in PCOS. |
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