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Vol 74, No 3 (2023)
Original paper
Submitted: 2022-10-18
Accepted: 2022-12-06
Published online: 2023-05-08
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MicroRNA-646 inhibits the proliferation of ovarian granulosa cells via insulin-like growth factor 1 (IGF-1) in polycystic ovarian syndrome (PCOS)

Jiali Lu1, Feilan Xuan2, Aixue Chen3, Ruiying Jin4, Weimei Zhou4, Yongju Ye5, Yuefang Ren1
DOI: 10.5603/EP.a2023.0020
·
Pubmed: 37155307
·
Endokrynol Pol 2023;74(3):305-314.
Affiliations
  1. Department of Gynaecology, Huzhou Maternity and Child Health Care Hospital, Huzhou, China
  2. Department of Obstetrics and Gynaecology, Hangzhou Traditional Chinese Medical Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou, China
  3. Department of Gynaecology, Changxing People’s Hospital of Chongming District, Shanghai, China
  4. Department of Ultrasound, Jiaojiang Maternal and Child Health Hospital, Taizhou, China
  5. Department of Gynaecology, Lishui Hospital of Traditional Chinese Medicine, Lishui, China

open access

Vol 74, No 3 (2023)
Original Paper
Submitted: 2022-10-18
Accepted: 2022-12-06
Published online: 2023-05-08

Abstract

Introduction: Polycystic ovarian syndrome (PCOS) is a common endocrinopathy in women. MicroRNAs (miRNAs) have been proven to play a crucial role in balancing the proliferation and apoptosis of granulosa cells (GCs) in PCOS.

Material and methods: The miRNA of PCOS was screened by bioinformatics analysis, and microRNA 646 (miR-646) was found to be involved in insulin-related pathways by enrichment analysis. The cell counting kit-8 (CCK-8), cell colony formation, and the 5-ethynyl-2’-deoxyuridine (EdU) assays were used to explore the effect of miR-646 on proliferation of GCs, flow cytometry was used to measure the cell cycle and apoptosis, and Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to explore the biological mechanism of miR-646. The human ovarian granulosa cells KGN were selected by measuring the miR-646 and via insulin-like growth factor 1 (IGF-1) levels and used for cell transfection.

Results: Overexpressed miR-646 inhibited KGN cell proliferation, and silenced miR-646 advanced it. Most cells were arrested in the S phase of cell cycle with overexpressed-miR-646, while after silencing miR-646, cells were arrested in the G2/M phase. And the miR-646 mimic raised apoptosis in KGN cells. Also, a dual-luciferase reporter proved the regulation effect of miR-646 on IGF-1, miR-646 mimic inhibited IGF-1, and miR-646 inhibitor advanced IGF-1. The cyclin D1, cyclin-dependent kinase 2 (CDK2), and B-cell CLL/lymphoma 2 (Bcl-2) levels were inhibited with overexpressed-miR-646, while silenced-miR-646 promoted their expression, and the bcl-2-like protein 4 (Bax) level was the opposite. This study found that silenced-IGF1 antagonized the promotive effect of the miR-646 inhibitor on cell proliferation.

Conclusions: MiR-646 inhibitor treatment can promote the proliferation of GCs by regulating the cell cycle and inhibiting apoptosis, while silenced-IGF-1 antagonizes it.

Abstract

Introduction: Polycystic ovarian syndrome (PCOS) is a common endocrinopathy in women. MicroRNAs (miRNAs) have been proven to play a crucial role in balancing the proliferation and apoptosis of granulosa cells (GCs) in PCOS.

Material and methods: The miRNA of PCOS was screened by bioinformatics analysis, and microRNA 646 (miR-646) was found to be involved in insulin-related pathways by enrichment analysis. The cell counting kit-8 (CCK-8), cell colony formation, and the 5-ethynyl-2’-deoxyuridine (EdU) assays were used to explore the effect of miR-646 on proliferation of GCs, flow cytometry was used to measure the cell cycle and apoptosis, and Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to explore the biological mechanism of miR-646. The human ovarian granulosa cells KGN were selected by measuring the miR-646 and via insulin-like growth factor 1 (IGF-1) levels and used for cell transfection.

Results: Overexpressed miR-646 inhibited KGN cell proliferation, and silenced miR-646 advanced it. Most cells were arrested in the S phase of cell cycle with overexpressed-miR-646, while after silencing miR-646, cells were arrested in the G2/M phase. And the miR-646 mimic raised apoptosis in KGN cells. Also, a dual-luciferase reporter proved the regulation effect of miR-646 on IGF-1, miR-646 mimic inhibited IGF-1, and miR-646 inhibitor advanced IGF-1. The cyclin D1, cyclin-dependent kinase 2 (CDK2), and B-cell CLL/lymphoma 2 (Bcl-2) levels were inhibited with overexpressed-miR-646, while silenced-miR-646 promoted their expression, and the bcl-2-like protein 4 (Bax) level was the opposite. This study found that silenced-IGF1 antagonized the promotive effect of the miR-646 inhibitor on cell proliferation.

Conclusions: MiR-646 inhibitor treatment can promote the proliferation of GCs by regulating the cell cycle and inhibiting apoptosis, while silenced-IGF-1 antagonizes it.

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Keywords

polycystic ovarian syndrome; granulosa cells; microRNA; miR-646; IGF1; KGN cells

About this article
Title

MicroRNA-646 inhibits the proliferation of ovarian granulosa cells via insulin-like growth factor 1 (IGF-1) in polycystic ovarian syndrome (PCOS)

Journal

Endokrynologia Polska

Issue

Vol 74, No 3 (2023)

Article type

Original paper

Pages

305-314

Published online

2023-05-08

Page views

1499

Article views/downloads

441

DOI

10.5603/EP.a2023.0020

Pubmed

37155307

Bibliographic record

Endokrynol Pol 2023;74(3):305-314.

Keywords

polycystic ovarian syndrome
granulosa cells
microRNA
miR-646
IGF1
KGN cells

Authors

Jiali Lu
Feilan Xuan
Aixue Chen
Ruiying Jin
Weimei Zhou
Yongju Ye
Yuefang Ren

References (33)
  1. Goodarzi MO, Dumesic DA, Chazenbalk G, et al. Polycystic ovary syndrome: etiology, pathogenesis and diagnosis. Nat Rev Endocrinol. 2011; 7(4): 219–231.
    CrossRefPubMed
  2. Anagnostis P, Tarlatzis BC, Kauffman RP. Polycystic ovarian syndrome (PCOS): Long-term metabolic consequences. Metabolism. 2018; 86: 33–43.
    CrossRefPubMed
  3. Wolf WM, Wattick RA, Kinkade ON, et al. Geographical Prevalence of Polycystic Ovary Syndrome as Determined by Region and Race/Ethnicity. Int J Environ Res Public Health. 2018; 15(11).
    CrossRefPubMed
  4. Madnani N, Khan K, Chauhan P, et al. Polycystic ovarian syndrome. Indian J Dermatol Venereol Leprol. 2013; 79(3): 310–321.
    CrossRefPubMed
  5. Escobar-Morreale HF. Polycystic ovary syndrome: definition, aetiology, diagnosis and treatment. Nat Rev Endocrinol. 2018; 14(5): 270–284.
    CrossRefPubMed
  6. Zeng X, Xie YJ, Liu YT, et al. Polycystic ovarian syndrome: Correlation between hyperandrogenism, insulin resistance and obesity. Clin Chim Acta. 2020; 502: 214–221.
    CrossRefPubMed
  7. Havelock JC, Rainey WE, Carr BR. Ovarian granulosa cell lines. Mol Cell Endocrinol. 2004; 228(1-2): 67–78.
    CrossRefPubMed
  8. Raei Sadigh A, Darabi M, Salmassi A, et al. Fractalkine and apoptotic/anti-apoptotic markers in granulosa cells of women with polycystic ovarian syndrome. Mol Biol Rep. 2020; 47(5): 3593–3603.
    CrossRefPubMed
  9. Chahal N, Geethadevi A, Kaur S, et al. Direct impact of gonadotropins on glucose uptake and storage in preovulatory granulosa cells: Implications in the pathogenesis of polycystic ovary syndrome. Metabolism. 2021; 115: 154458.
    CrossRefPubMed
  10. Gong Y, Luo S, Fan P, et al. Growth hormone activates PI3K/Akt signaling and inhibits ROS accumulation and apoptosis in granulosa cells of patients with polycystic ovary syndrome. Reprod Biol Endocrinol. 2020; 18(1): 121.
    CrossRefPubMed
  11. Li M, Zhao H, Zhao SG, et al. The HMGA2-IMP2 Pathway Promotes Granulosa Cell Proliferation in Polycystic Ovary Syndrome. J Clin Endocrinol Metab. 2019; 104(4): 1049–1059.
    CrossRefPubMed
  12. Ji XL, Liu X, Wang Z, et al. Expression of ARID1A in polycystic ovary syndrome and its effect on the proliferation and apoptosis of ovarian granulosa cells. Ann Endocrinol (Paris). 2020; 81(6): 521–529.
    CrossRefPubMed
  13. Chen B, Xu P, Wang J, et al. The role of MiRNA in polycystic ovary syndrome (PCOS). Gene. 2019; 706: 91–96.
    CrossRefPubMed
  14. Bao D, Li M, Zhou D, et al. miR-130b-3p is high-expressed in polycystic ovarian syndrome and promotes granulosa cell proliferation by targeting SMAD4. J Steroid Biochem Mol Biol. 2021; 209: 105844.
    CrossRefPubMed
  15. Jiang YC, Ma JX. The role of MiR-324-3p in polycystic ovary syndrome (PCOS) via targeting WNT2B. Eur Rev Med Pharmacol Sci. 2018; 22(11): 3286–3293.
    CrossRefPubMed
  16. Sun D, Liu J, Zhou L. Upregulation of circular RNA circ‑FAM53B predicts adverse prognosis and accelerates the progression of ovarian cancer via the miR‑646/VAMP2 and miR‑647/MDM2 signaling pathways. Oncol Rep. 2019; 42(6): 2728–2737.
    CrossRefPubMed
  17. Zhang P, Tang WM, Zhang H, et al. MiR-646 inhibited cell proliferation and EMT-induced metastasis by targeting FOXK1 in gastric cancer. Br J Cancer. 2017; 117(4): 525–534.
    CrossRefPubMed
  18. Yuan X, Liu Y, Chen E, et al. MiR-646 regulates proliferation and migration of laryngeal carcinoma through the PI3K/AKT pathway via targeting GPX1. Oral Dis. 2021; 27(7): 1678–1686.
    CrossRefPubMed
  19. Wang Bo, Lu Ye, Feng E. hsa_circ_0001610 knockdown modulates miR-646-STAT3 axis to suppress endometrial carcinoma progression. J Gene Med. 2021; 23(6): e3337.
    CrossRefPubMed
  20. Farhadi-Azar M, Ghahremani M, Mahboobifard F, et al. Effects of Rosa damascena on reproductive improvement, metabolic parameters, liver function and insulin-like growth factor-1 gene expression in estradiol valerate induced polycystic ovarian syndrome in Wistar rats. Biomed J. 2022 [Epub ahead of print].
    CrossRefPubMed
  21. Sirotkin A, Alexa R, Kádasi A, et al. Resveratrol directly affects ovarian cell sirtuin, proliferation, apoptosis, hormone release and response to follicle-stimulating hormone (FSH) and insulin-like growth factor I (IGF-I). Reprod Fertil Dev. 2019 [Epub ahead of print].
    CrossRefPubMed
  22. Yang J, Zhou J, Cui B, et al. Evaluation of Hypoxia on the Expression of miR-646/IGF-1 Signaling in Human Periodontal Ligament Cells (hPDLCs). Med Sci Monit. 2018; 24: 5282–5291.
    CrossRefPubMed
  23. Wang Z. The guideline of the design and validation of MiRNA mimics. Methods Mol Biol. 2011; 676: 211–223.
    CrossRefPubMed
  24. Zhang Z, Li J, Huang Y, et al. Upregulated miR-1258 regulates cell cycle and inhibits cell proliferation by directly targeting E2F8 in CRC. Cell Prolif. 2018; 51(6): e12505.
    CrossRefPubMed
  25. Zheng Q, Li Y, Zhang D, et al. ANP promotes proliferation and inhibits apoptosis of ovarian granulosa cells by NPRA/PGRMC1/EGFR complex and improves ovary functions of PCOS rats. Cell Death Dis. 2017; 8(10): e3145.
    CrossRefPubMed
  26. Guo M, Lv M, Shao Y, et al. Bax functions as coelomocyte apoptosis regulator in the sea cucumber Apostichopus japonicus. Dev Comp Immunol. 2020; 102: 103490.
    CrossRefPubMed
  27. Alenzi FQB. Links between apoptosis, proliferation and the cell cycle. Br J Biomed Sci. 2004; 61(2): 99–102.
    CrossRefPubMed
  28. O'Connor MJ, Thakar T, Nicolae CM, et al. PARP14 regulates cyclin D1 expression to promote cell-cycle progression. Oncogene. 2021; 40(30): 4872–4883.
    CrossRefPubMed
  29. Miyashita S, Owa T, Seto Y, et al. Cyclin D1 controls development of cerebellar granule cell progenitors through phosphorylation and stabilization of ATOH1. EMBO J. 2021; 40(14): e105712.
    CrossRefPubMed
  30. Spencer SL, Cappell SD, Tsai FC, et al. The proliferation-quiescence decision is controlled by a bifurcation in CDK2 activity at mitotic exit. Cell. 2013; 155(2): 369–383.
    CrossRefPubMed
  31. Zhang J, Xu Y, Liu H, et al. MicroRNAs in ovarian follicular atresia and granulosa cell apoptosis. Reprod Biol Endocrinol. 2019; 17(1): 9.
    CrossRefPubMed
  32. Han Y, Wang S, Wang Y, et al. IGF-1 Inhibits Apoptosis of Porcine Primary Granulosa Cell by Targeting Degradation of Bim. Int J Mol Sci. 2019; 20(21).
    CrossRefPubMed
  33. Dou L, Zheng Y, Li Lu, et al. The effect of cinnamon on polycystic ovary syndrome in a mouse model. Reprod Biol Endocrinol. 2018; 16(1): 99.
    CrossRefPubMed

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