Expression And Clinical Significance Of MiR-93 In The Serum Of Patients With Polycystic Ovary Syndrome

Objective:Polycystic ovary syndrome(PCOS)is a common endocrine and metabolic disease for childbearing-age women,which features hyperandrogenemia,ovulation dysfunction and insulin resistance.PCOS can cause amenorrhea,infrequent menstruation,infertility and polycystic ovarian manifestations,also it is the crucial factor of type 2 diabetes,cardiovascular disease and endometrial cancer,with other metabolic disorders such as obesity,insulin resistance and dyslipidemia.It does harmful to the patients' lives.MicroRNA(miRNA)refers to small RNA molecules with gene regulation function,which works through controlling target gene mRNA 3 'untranslated region(UTR)and further blocking degradation or translation of target gene mRNA.Recent studies have shown close relationship between the differential expression of miRNA and the development of PCOS.This study detected the change of miR-93 expression in PCOS patients and analyzed the relationship between the change of miR-93 and PCOS clinical features,and discussed about the clinical meaning of miR-93 in the development of PCOS.Furthermore,we examined the effect of miR-93 on the proliferation of human ovarian granular cell line KGN cells,and identified the target gene of miR-93.This research aims to providing further basis for syndrome pathogenesis and clinical treatment of PCOS.Methods:We collected the serum samples from 25 PCOS patients and also took serum of patients with normal ovarian function as the control group(n=25).The age,height,weight,six hormones(including follicle-stimulating hormone,luteinizing hormone,estrogen,progesterone,testosterone and prolactin),serum insulin concentration and serum glucose concentration were recorded and compared.SYBR Green I real-time quantitative PCR was used to detect the relative expression level of miR-93 in serum.The relationship between the relative level of miR-93 and clinical features of polycystic ovary syndrome was analyzed further.To evaluate the diagnostic value of miR-93 in the polycystic ovary syndrome,ROC curve was plotted and the AUC was calculated.We detected the growth and proliferation of human granulosa-like tumor cell line KGN cells using MTT and colony formation.Moreover,we predicted the target gene of miR-93 through bioinformatics,and verified the direct target gene of miR-93 by the double luciferase in the human granulosa-like tumor cell line KGN.SPSS17.0 was used;P < 0.05 showed that the difference was statistically significant.Results:Through the analysis of data from PCOS patients and normal ovarian function patients,we found that PCOS patients,the BMI,luteinizing hormone level,testosterone level,insulin concentration and serum glucose level were significantly higher(P > 0.05).The relative expression level of miR-93 in patients with polycystic ovary syndrome was significantly increased(Z=7.614,P<0.01).In the control group,the correlation coefficients(r value)of miR-93 with testosterone concentration,insulin concentration,and insulin resistance(HOMA-IR)were 0.03,0.42,and 0.39 respectively(P=0.95,0.11,0.13).In the PCOS group,the r value of miR-93 with testosterone concentration,insulin concentration,and insulin resistance(HOMA-IR)were 0.33,0.24,and 0.28 respectively(P=0.09,0.41,0.47).ROC curve showed that AUC area is 0.73,with 70% sensitivity and 78% specificity,also the 95% confidence interval is [0.59-0.87].In human granulosa-like tumor cell line KGN,compared with the ASO control group,the growth activity and the proliferating rate of the cells were significantly decreased after closing the expression of miR-93;however,the growth activity and the proliferating ability of the cells were significantly increased after over-expression of miR-93.The results of luciferase experiment showed that miR-93 can target RAB5 B mRNA 3' UTR and inhibit the activity of luciferase.In human granulosa-like tumor cell line KGN,the levels of RAB5 B mRNA and protein were significantly increased after transfection with miR-93 ASO,while we got the opposite results for KGN cells transfected with miR-93 mimics.Conclusions:1.The relative serum level of miR-93 in patients with PCOS is significantly high,and there was no correlation between miR-93 with any of the metabolic and hormonal parameters including insulin,insulin resistance(HOMA-IR)or testosterone.2.The serum of miR-93 for diagnosing PCOS is more accuracy,and maybe has much value for the diagnosing of PCOS.3.The miR-93 can promote the growth and proliferation of human granulosa-like tumor cell line KGN cells.4.RAB5 B is a direct target gene of miR-93 and negatively regulated by it.