MicroRNA-135a Regulates VEGFC Expression and Promotes Luteinized Granulosa Cell Apoptosis in Polycystic Ovary Syndrome


Androgen is known to regulate microRNA-135a (miR-135a) and can be regulated by androgen, suggesting that it may contribute to polycystic ovary syndrome (PCOS) with hyperandrogenism. However, its roles and mechanisms of action in PCOS are unknown. In this study, the role and molecular mechanisms underlying miR-135a in granulosa cells (GCs) in PCOS were evaluated. miR-135a expression was upregulated in patients with PCOS and in GCs isolated from patients compared with that in the respective controls (P < 0.01), as determined by RT-qPCR. The overexpression of miR-135a inhibited GC proliferation and induced GC apoptosis, as observed by CCK-8 assay and apoptosis assay. Furthermore, miR-135a overexpression increased the expression of double-strand break maker, γH2AX, as confirmed by western blotting. Our results further suggest that these effects were mediated via downregulation of vascular endothelial growth factor C (VEGFC), which was identified as a direct target of miR-135a. Moreover, levels of VEGFC and miR-135a expression showed a negative correlation. These findings indicate that miR-135a promotes apoptosis and the DNA damage response in GCs in PCOS, likely via VEGFC signaling. This study provides novel insights into GC dysregulation in PCOS and suggests that miR-135a is a promising therapeutic target for PCOS treatment.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. 1.

    Glintborg D, Andersen M. An update on the pathogenesis, inflammation, and metabolism in hirsutism and polycystic ovary syndrome. Gynecol Endocrinol. 2010;26:281–96.

    CAS  Article  Google Scholar 

  2. 2.

    ESHRE TR, Group APCW. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril. 2004;81:19–25.

    Google Scholar 

  3. 3.

    Yildiz BO, Bozdag G, Yapici Z, Esinler I, Yarali H. Prevalence, phenotype and cardiometabolic risk of polycystic ovary syndrome under different diagnostic criteria. Hum Reprod. 2012;27:3067–73.

    Article  Google Scholar 

  4. 4.

    Maciel GA, Baracat EC, Benda JA, Markham SM, Hensinger K, Chang RJ, et al. Stockpiling of transitional and classic primary follicles in ovaries of women with polycystic ovary syndrome. J Clin Endocrinol Metab. 2004;89:5321–7.

    CAS  Article  Google Scholar 

  5. 5.

    Jahromi BN, Mosallanezhad Z, Matloob N, Davari M, Ghobadifar MA. The potential role of granulosa cells in the maturation rate of immature human oocytes and embryo development: a co-culture study. Clinical and experimental reproductive medicine. 2015;42:111–7.

    Article  Google Scholar 

  6. 6.

    Homburg R. Involvement of growth factors in the pathophysiology of polycystic ovary syndrome. Gynecol Endocrinol. 1998;12:391–7.

    CAS  Article  Google Scholar 

  7. 7.

    Tiwari M, Prasad S, Tripathi A, Pandey AN, Ali I, Singh AK, et al. Apoptosis in mammalian oocytes: a review. APOPTOSIS. 2015;20:1019–25.

    CAS  Article  Google Scholar 

  8. 8.

    Erickson GF, Magoffin DA, Garzo VG, Cheung AP, Chang RJ. Granulosa cells of polycystic ovaries: are they normal or abnormal? Hum Reprod. 1992;7:293–9.

    CAS  Article  Google Scholar 

  9. 9.

    Willis DS, Watson H, Mason HD, Galea R, Brincat M, Franks S. Premature response to luteinizing hormone of granulosa cells from anovulatory women with polycystic ovary syndrome: relevance to mechanism of anovulation. J Clin Endocrinol Metab. 1998;83:3984–91.

    CAS  PubMed  Google Scholar 

  10. 10.

    Ambros V. MicroRNAs: tiny regulators with great potential. CELL. 2001;107:823–6.

    CAS  Article  Google Scholar 

  11. 11.

    Wen-Na Yuan LT. MicroRNA320 inhibits insulin resistance in patients with PCOS through regulating erk12 signaling pathway. BIOMED RES-TOKYO. 2017;28:4946–9.

    Google Scholar 

  12. 12.

    Fu X, He Y, Wang X, Peng D, Chen X, Li X, et al. MicroRNA-16 promotes ovarian granulosa cell proliferation and suppresses apoptosis through targeting PDCD4 in polycystic ovarian syndrome. Cell Physiol Biochem. 2018;48:670–82.

    CAS  Article  Google Scholar 

  13. 13.

    Anja Elaine Sørensen MLWS, Dalgaard ALT. MicroRNAs related to polycystic ovary syndrome (PCOS). GENES-BASEL. 2014;5:684–708.

    Article  Google Scholar 

  14. 14.

    Cho S, Mutlu L, Grechukhina O, Taylor HS. Circulating microRNAs as potential biomarkers for endometriosis. Fertil Steril. 2015;103:1252–60.

    CAS  Article  Google Scholar 

  15. 15.

    Wan X, Pu H, Huang W, Yang S, Zhang Y, Kong Z, et al. Androgen-induced miR-135a acts as a tumor suppressor through downregulating RBAK and MMP11, and mediates resistance to androgen deprivation therapy. Oncotarget. 2016;7:51284–300.

    Article  Google Scholar 

  16. 16.

    Kroiss A, Vincent S, Decaussin-Petrucci M, Meugnier E, Viallet J, Ruffion A, et al. Androgen-regulated microRNA-135a decreases prostate cancer cell migration and invasion through downregulating ROCK1 and ROCK2. ONCOGENE. 2015;34:2846–55.

    CAS  Article  Google Scholar 

  17. 17.

    Catteau-Jonard S, Jamin SP, Leclerc A, Gonzales J, Dewailly D, di Clemente N. Anti-Mullerian hormone, its receptor, FSH receptor, and androgen receptor genes are overexpressed by granulosa cells from stimulated follicles in women with polycystic ovary syndrome. J Clin Endocrinol Metab. 2008;93:4456–61.

    CAS  Article  Google Scholar 

  18. 18.

    Wei Y, Lai X, Yu S, et al. Exosomal miR-221/222 enhances tamoxifen resistance in recipient ER-positive breast cancer cells. Breast Cancer Res Treat. 2014;147:423–31.

    CAS  Article  Google Scholar 

  19. 19.

    Roos WP, Kaina B. DNA damage-induced cell death by apoptosis. Trends Mol Med. 2006;12:440–50.

    CAS  Article  Google Scholar 

  20. 20.

    Hughesdon PE. Morphology and morphogenesis of the Stein-Leventhal ovary and of so-called “hyperthecosis”. OBSTET GYNECOL SURV. 1982;37:59–77.

    CAS  Article  Google Scholar 

  21. 21.

    Lin F, Li R, Pan ZX, et al. miR-26b promotes granulosa cell apoptosis by targeting ATM during follicular atresia in porcine ovary. PLoS One. 2012;7:e38640.

    CAS  Article  Google Scholar 

  22. 22.

    Chen Y, Heneidi S, Lee J, Layman LC, Stepp DW, Gamboa GM, et al. miRNA-93 inhibits GLUT4 and is overexpressed in adipose tissue of polycystic ovary syndrome patients and women with insulin resistance. DIABETES. 2013;62:2278–86.

    CAS  Article  Google Scholar 

  23. 23.

    Dokras A, Sarwer DB, Allison KC, Milman L, Kris-Etherton PM, Kunselman AR, et al. Weight loss and lowering androgens predict improvements in health-related quality of life in women with PCOS. J Clin Endocrinol Metab. 2016;101:2966–74.

    CAS  Article  Google Scholar 

  24. 24.

    Hominick D, Silva A, Khurana N, et al. VEGF-C promotes the development of lymphatics in bone and bone loss. ELIFE. 2018;7:e34323.

    Article  Google Scholar 

  25. 25.

    Fang S, Nurmi H, Heinolainen K, Chen S, Salminen E, Saharinen P, et al. Critical requirement of VEGF-C in transition to fetal erythropoiesis. BLOOD. 2016;128:710–20.

    CAS  Article  Google Scholar 

  26. 26.

    Zhao L, Zhu Z, Yao C, Huang Y, Zhi E, Chen H, et al. VEGFC/VEGFR3 signaling regulates mouse spermatogonial cell proliferation via the activation of AKT/MAPK and cyclin D1 pathway and mediates the apoptosis by affecting caspase 3/9 and Bcl-2. Cell Cycle. 2018;17:225–39.

    CAS  Article  Google Scholar 

  27. 27.

    Zheng Q, Zhou F, Cui X, Liu M, Li Y, Liu S, et al. Novel serum biomarkers detected by protein array in polycystic ovary syndrome with low progesterone level. Cell Physiol Biochem. 2018;46:2297–310.

    CAS  Article  Google Scholar 

Download references


This work was supported in part by grants from the Shanghai Commission of Science and Technology (17DZ2271100), Program of Shanghai Academic Research Leader in Shanghai Municipal Commission of Health and Family Planning (No. 2017BR015), Shanghai Technological Innovation Plan (No. 18140902400), National Natural Science Foundation of China (No. 81901442), and Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant Support (No. 20161413).

Author information




Yifang Wei and Yu Hu designed and performed experiments and wrote the original draft manuscript, Shenglian Lu collected the clinical samples, Li Guo and Xiaoyu Wu analyzed the data, Xing Liu performed experiments, and Yun Sun oversaw the entire study and edited the manuscript.

Corresponding authors

Correspondence to Yifang Wei or Yun Sun.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(DOC 33 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wei, Y., Lu, S., Hu, Y. et al. MicroRNA-135a Regulates VEGFC Expression and Promotes Luteinized Granulosa Cell Apoptosis in Polycystic Ovary Syndrome. Reprod. Sci. 27, 1436–1442 (2020). https://doi.org/10.1007/s43032-020-00155-0

Download citation


  • miR-135a
  • PCOS
  • Apoptosis