多囊卵巢综合征患者卵丘颗粒细胞中特异性环状RNA的表达研究

Novel circular RNAs expression in cumulus granulosa cells of patients with polycystic ovary syndrome

目的研究多囊卵巢综合征(PCOS)患者卵丘颗粒细胞中特异性环状RNA(circRNA)的表达,探讨circRNA在诊断PCOS中的意义。方法收集2017年5~12月期间于烟台毓璜顶医院生殖医学科行IVF-ET/ICSI助孕的6名典型PCOS患者和6名卵巢正常对照者的卵丘颗粒细胞进行circRNA芯片检测,另收集同期行IVF-ET/ICSI助孕的PCOS及正常对照者的卵丘颗粒细胞各25例进行实时荧光定量PCR(qRT-PCR)验证芯片结果,记录两组患者的基础内分泌及促排卵情况等临床资料,Spearman秩相关分析差异表达的circRNA与PCOS患者临床特征资料之间的相关性,并绘制受试者工作特征曲线(ROC)评估circRNA对PCOS的诊断效力。结果circRNA芯片共检测出286个差异表达的circRNA(与正常对照相比,PCOS患者卵丘颗粒细胞中分别有167个上调,119个下调),经后续挑选和qRT-PCR验证,hsa_circ_0043533在PCOS组中显著高表达(P<0.05),而hsa_circ_0097636显著低表达(P<0.01)。Spearman秩相关分析提示PCOS患者血睾酮(T)值与hsa_circ_0043533和hsa_circ_0097636表达水平均存在正相关性(分别为r=0.3999,P=0.0476;r=0.5075,P=0.0096)。ROC曲线分析显示,联合hsa_circ_0097636和T诊断PCOS的曲线下面积(AUC)显著高于单独采用hsa_circ_0043533(0.893vs.0.709)或hsa_circ_0097636(0.893vs.0.738)。结论circRNA在PCOS及正常患者卵丘颗粒细胞中的表达存在显著差异,经qRT-PCR验证出的两个circRNA可能为深入理解PCOS发病机制和探索诊治方法提供更为广阔的思路。

Objective:To investigate the expression and potential significance of circular RNA(circRNA)in the cumulus granulosa cells of patients with polycystic ovary syndrome(PCOS).Methods:The cumulus granulosa cells were collected from 6 PCOS patients and 6 normal control individuals for microarray analysis in the Center for Reproductive Medicine,Yantai Yuhuangding Hospital from May to Dec.2017.In addition,the cumulus granulosa cells were collected from 25 PCOS patients and 25 normal controls assisted by IVF-ET/ICSI for real-time quantitative fluorescence PCR(qRT-PCR)to verify the chip results during the same period.All patients’clinical characteristics including the basal hormone levels and ovarian stimulation were collected.Spearman’s rank correlation and receiver operating characteristic(ROC)were performed to analyze the potential correlation between novel circRNA and patients’clinical characteristics and evaluate the diagnostic efficacy of circRNA for PCOS.Results:A total of 286 differential expression of circRNA were identified by microarray in PCOS group.Compared with normal controls,there were 167 up-regulation and 119 down-regulation of expressions of circRNA in cumulus granule cells of PCOS patients.The further qRT-PCR validation found that expression level of hsa_circ_0043533 was significantly higher(P<0.05)and hsa_circ_0097636 was significantly lower in PCOS group(P<0.01).Spearman’s rank correlation analysis indicated that serum testosterone(T)level was positively correlated with the expression of hsa_circ_0043533 and hsa_circ_0097636 in PCOS group(r=0.399 9,P=0.047 6 and r=0.507 5,P=0.009 6,respectively).The ROC curve analysis showed that the combination of hsa_circ_0097636 and T as diagnostic indicator for PCOS resulted in a larger area under the curve(AUC)than hsa_circ_0043533(0.893 vs.0.709)or hsa_circ_0097636(0.893 vs.0.738)alone.Conclusions:The circRNA expressions in cumulus granulosa cells are significantly different between PCOS and normal control individuals.Two circRNAs identified by qRT-PCR may offer great promises in understanding the pathogenesis and developing effective therapeutic interventions for PCOS patients.