摘要
目的通过生物信息学方法探索与多囊卵巢综合征(PCOS)发病相关的基因并在卵巢颗粒细胞中进行验证,为筛选PCOS潜在分子标志物提供参考。方法分别以GSE34526和GSE137684数据集作为训练集和验证集,通过差异表达分析和加权基因共表达网络分析探索与PCOS发病潜在相关的炎症相关基因,随后通过受试者工作特征(ROC)曲线对其预测价值进行验证。选取在右江民族医学院附属医院就诊的PCOS患者30例(PCOS组)与非PCOS患者27例(对照组)为研究对象,应用RT-qPCR验证颗粒细胞中核心基因的表达水平,采用Spearman相关和多元线性回归分析核心基因表达量与各临床参数的相关性,并以ROC曲线分析核心基因表达量对PCOS的诊断效能,最后在动物模型上验证核心基因的蛋白表达水平。结果共鉴定出1888个差异表达基因、89个模块相关基因和366个炎症反应相关基因,通过取共表达基因及ROC曲线验证最终获得核心基因磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基γ(PIK3CG)。RT-qPCR结果显示PCOS组颗粒细胞中的PIK3CG表达量高于对照组(P<0.05)。Spearman相关分析显示,PIK3CG表达水平与BMI、睾酮(T)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)呈正相关(P<0.05)。多元线性回归分析显示,PCOS患者的FINS、HOMA-IR、BMI随着PIK3CG mRNA相对表达水平的升高而增加(P<0.05)。ROC曲线分析显示,颗粒细胞中PIK3CG mRNA相对表达水平对诊断PCOS的ROC曲线下面积为0.6593。免疫组化结果可见PCOS组小鼠卵巢组织中的PIK3CG蛋白表达明显升高。结论PIK3CG可能与PCOS发病有一定的相关性。
Objective To explore the genes related to the pathogenesis of polycystic ovary syndrome(PCOS)through bioinformatics methods and verify them in ovarian granulosa cells,providing a reference for screening potential molecular markers of PCOS.Methods The GSE34526 and GSE137684 datasets were respectively used as the training set and validation set.The inflammation-related genes potentially associated with the onset of PCOS were explored through differential expression analysis and weighted gene co-expression network analysis.Subsequently,their predictive value was verified through the receiver operating characteristic(ROC)curve.Thirty patients with PCOS(the PCOS group)and 27 patients without PCOS(the control group)who were treated in Affiliated Hospital of Youjiang Medical University for Nationalities were selected as the research objects.RT-qPCR was applied to verify the expression level of core genes in granular cells.Spearman correlation and multiple linear regression were used to analyze the correlation between the expression level of core genes and various clinical parameters,and the ROC curve was used to analyze the diagnostic efficacy of the expression level of core genes for PCOS.Finally,the protein expression level of core genes was verified on animal models.Results A total of 1888 differentially expressed genes,89 module-related genes and 366 inflammatory response-related genes were identified.The core gene phosphatidylinositol-4,5-diphosphate 3-kinase catalytic subunit γ(PIK3CG)was ultimately obtained by taking co-expressed genes and verifying with the ROC curve.RT-qPCR results showed that the expression level of PIK3CG in granulosa cells of the PCOS group was higher than that of the control group(P<0.05).Spearman correlation analysis showed that the expression level of PIK3CG was positively correlated with BMI,testosterone(T),fasting insulin(FINS),and insulin resistance index(HOMA-IR,P<0.05).Multivariate linear regression analysis showed that FINS,HOMA-IR and BMI were increased with the increasing of PIK3CG expression level(P<0.05).ROC curve analysis showed that the area under the curve for diagnosing PCOS patients with PIK3CG mRNA expression levels in granulosa cells was 0.6593.The immunohistochemical results showed that the expression of PIK3CG protein in the ovarian tissues of mice in the PCOS group was significantly increased.Conclusion PIK3CG may be potentially associated with the pathogenesis of PCOS.
作者
黄利营
李丽宁
覃晓
凌少华
田纯
凌燕兰
陈荣
覃莉
HUANG Liying;LI Lining;QIN Xiao;LING Shaohua;TIAN Chun;LING Yanlan;CHEN Rong;QIN Li;△(Graduate School,Youjiang Medical University for Nationalities,Baise,Guangxi 533000,China;Reproductive Medicine Center,Baise People’s Hospital,Baise,Guangxi 533000,China;Reproductive Medicine Center,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise,Guangxi 533099,China)
出处
《重庆医学》
2025年第8期1771-1780,共10页
Chongqing Medical Journal
基金
广西自然科学基金项目(2019GXNSFBA245020)
广西壮族自治区卫生健康委自筹经费科研课题(Z-L20230901)
右江民族医学院附属医院高层次人才科研项目(Y20196316)。
作者简介
通信作者:覃莉,E-mail:qinlimy@126.com。
