Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis
Abstract Background Polycystic ovary syndrome (PCOS) pathogenesis involves dysregulated granulosa cell function, but molecular mechanisms remain unclear. Methods High-throughput RNA sequencing was performed on ovarian granulosa cells from 6 PCOS patients and 3 controls to identify differentially exp...
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BMC
2025-08-01
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| Series: | Journal of Ovarian Research |
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| Online Access: | https://doi.org/10.1186/s13048-025-01746-8 |
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| author | Ping Tao Xiaohong Yan Zhanxiang Wang |
| author_facet | Ping Tao Xiaohong Yan Zhanxiang Wang |
| author_sort | Ping Tao |
| collection | DOAJ |
| description | Abstract Background Polycystic ovary syndrome (PCOS) pathogenesis involves dysregulated granulosa cell function, but molecular mechanisms remain unclear. Methods High-throughput RNA sequencing was performed on ovarian granulosa cells from 6 PCOS patients and 3 controls to identify differentially expressed mRNAs. Bioinformatics analyses including ceRNA network construction predicted the KRTAP5-AS1/miR-199b-5p/CYP19A1 regulatory axis, which was experimentally validated through dual-luciferase reporter assays. qRT-PCR confirmed the expression patterns of these molecules in expanded clinical cohorts (38 PCOS vs. 30 controls), with Pearson correlation analysis examining relationships between gene expression and clinical parameters. Using the KGN granulosa cell line, functional studies included: (1) ELISA quantification of estradiol production; (2) proliferation assessment via CCK-8 and colony formation assays; and (3) apoptosis evaluation by flow cytometry and Bax/Bcl-2 protein analysis. These experiments were performed following both gain-of-function (overexpression) and loss-of-function (shRNA knockdown) manipulations of KRTAP5-AS1 and miR-199b-5p. Results Through RNA sequencing of ovarian granulosa cells from 6 PCOS patients and 3 controls, we identified CYP19A1 as significantly upregulated in PCOS. Expanded validation in 38 PCOS vs. 30 controls confirmed elevated CYP19A1 and reduced miR-199b-5p in PCOS, with KRTAP5-AS1 showing negative correlation to miR-199b-5p and positive to CYP19A1. Clinically, CYP19A1 upregulation correlated with poor embryo quality, elevated testosterone, AMH, BMI, and infertility duration, while miR-199b-5p levels associated positively with embryo quality. In KGN granulosa cells, miR-199b-5p overexpression suppressed CYP19A1 expression and estradiol synthesis, whereas KRTAP5-AS1 overexpression alleviated this suppression via competitive miR-199b-5p binding. Functional studies demonstrated that miR-199b-5p overexpression combined with KRTAP5-AS1 knockdown inhibited proliferation, promoted apoptosis, and reduced estradiol production, while opposite manipulations reversed these effects. Conclusions Our findings reveal that KRTAP5-AS1 modulates granulosa cell dysfunction in PCOS through the miR-199b-5p/CYP19A1 axis, highlighting miR-199b-5p as a potential therapeutic target for PCOS-related ovarian dysfunction and endocrine abnormalities. Clinical trial number Not applicable. |
| format | Article |
| id | doaj-art-fb24cbd0e42448cd928d2f036511864a |
| institution | Kabale University |
| issn | 1757-2215 |
| language | English |
| publishDate | 2025-08-01 |
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| series | Journal of Ovarian Research |
| spelling | doaj-art-fb24cbd0e42448cd928d2f036511864a2025-08-20T04:03:01ZengBMCJournal of Ovarian Research1757-22152025-08-0118111710.1186/s13048-025-01746-8Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesisPing Tao0Xiaohong Yan1Zhanxiang Wang2Department of Reproductive Medicine Centre, First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen UniversityDepartment of Reproductive Medicine Centre, First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen UniversityDepartment of Neurosurgery, First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen UniversityAbstract Background Polycystic ovary syndrome (PCOS) pathogenesis involves dysregulated granulosa cell function, but molecular mechanisms remain unclear. Methods High-throughput RNA sequencing was performed on ovarian granulosa cells from 6 PCOS patients and 3 controls to identify differentially expressed mRNAs. Bioinformatics analyses including ceRNA network construction predicted the KRTAP5-AS1/miR-199b-5p/CYP19A1 regulatory axis, which was experimentally validated through dual-luciferase reporter assays. qRT-PCR confirmed the expression patterns of these molecules in expanded clinical cohorts (38 PCOS vs. 30 controls), with Pearson correlation analysis examining relationships between gene expression and clinical parameters. Using the KGN granulosa cell line, functional studies included: (1) ELISA quantification of estradiol production; (2) proliferation assessment via CCK-8 and colony formation assays; and (3) apoptosis evaluation by flow cytometry and Bax/Bcl-2 protein analysis. These experiments were performed following both gain-of-function (overexpression) and loss-of-function (shRNA knockdown) manipulations of KRTAP5-AS1 and miR-199b-5p. Results Through RNA sequencing of ovarian granulosa cells from 6 PCOS patients and 3 controls, we identified CYP19A1 as significantly upregulated in PCOS. Expanded validation in 38 PCOS vs. 30 controls confirmed elevated CYP19A1 and reduced miR-199b-5p in PCOS, with KRTAP5-AS1 showing negative correlation to miR-199b-5p and positive to CYP19A1. Clinically, CYP19A1 upregulation correlated with poor embryo quality, elevated testosterone, AMH, BMI, and infertility duration, while miR-199b-5p levels associated positively with embryo quality. In KGN granulosa cells, miR-199b-5p overexpression suppressed CYP19A1 expression and estradiol synthesis, whereas KRTAP5-AS1 overexpression alleviated this suppression via competitive miR-199b-5p binding. Functional studies demonstrated that miR-199b-5p overexpression combined with KRTAP5-AS1 knockdown inhibited proliferation, promoted apoptosis, and reduced estradiol production, while opposite manipulations reversed these effects. Conclusions Our findings reveal that KRTAP5-AS1 modulates granulosa cell dysfunction in PCOS through the miR-199b-5p/CYP19A1 axis, highlighting miR-199b-5p as a potential therapeutic target for PCOS-related ovarian dysfunction and endocrine abnormalities. Clinical trial number Not applicable.https://doi.org/10.1186/s13048-025-01746-8KRTAP5-AS1miR-199b-5pCYP19A1Polycystic ovary syndromeGranulosa cell dysfunction |
| spellingShingle | Ping Tao Xiaohong Yan Zhanxiang Wang Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis Journal of Ovarian Research KRTAP5-AS1 miR-199b-5p CYP19A1 Polycystic ovary syndrome Granulosa cell dysfunction |
| title | Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis |
| title_full | Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis |
| title_fullStr | Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis |
| title_full_unstemmed | Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis |
| title_short | Mechanistic role of the KRTAP5-AS1/miR-199b-5p/CYP19A1 axis in polycystic ovary syndrome pathogenesis |
| title_sort | mechanistic role of the krtap5 as1 mir 199b 5p cyp19a1 axis in polycystic ovary syndrome pathogenesis |
| topic | KRTAP5-AS1 miR-199b-5p CYP19A1 Polycystic ovary syndrome Granulosa cell dysfunction |
| url | https://doi.org/10.1186/s13048-025-01746-8 |
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