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Daidzein Activates Akt Pathway to Promote the Proliferation of Female Germline Stem Cells through Upregulating Clec11a

Stem Cell Reviews and Reports volume 18, pages 3021–3032 (2022)Cite this article

Abstract

Female germline stem cells (FGSCs) have been successfully isolated and characterized from postnatal mammalian and human ovarian tissues. However, the effects and mechanisms of action of natural small-molecule compounds on FGSCs are largely unknown. Here, we found that daidzein promoted the viability and proliferation of FGSCs. To elucidate the mechanism underlying this, we performed RNA-Sequence in daidzein-treated FGSCs and controls. The results showed that there were 153 upregulated and 156 downregulated genes in daidzein treatment. We confirmed the expression of some genes related to cell proliferation in the sequencing results by RT-PCR, such as Type C lectin domain family 11 member a (Clec11a), Mucin1 (Muc1), Glutathione peroxidase 3 (Gpx3), and Tet methylcytosine dioxygenase 1 (Tet1). The high expression of Clec11a at the protein level after daidzein treatment was also confirmed by western blotting. Furthermore, recombinant mouse Clec11a (rmClec11a) protein was shown to promote the viability and proliferation of FGSCs. However, knockdown of Clec11a inhibited the viability and proliferation of FGSCs, which could not be rescued by the administration of daidzein. These results indicate that daidzein promoted the viability and proliferation of FGSCs through Clec11a. In addition, both daidzein and rmClec11a activated the Akt signaling pathway in FGSCs. However, Clec11a knockdown inhibited this pathway, which could not be rescued by daidzein administration. Taken together, our findings revealed that daidzein activates the Akt signaling pathway to promote cell viability and proliferation through upregulating Clec11a. This study should deepen our understanding of the developmental mechanism of FGSCs and female infertility.

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Data Availability

Availability of Data and Material Raw sequencing reads were obtained from Gene Expression Omnibus and processed with the parameters detailed below: GSE199120.

Code Availability

Not applicable.

Abbreviations

FGSC:

Female germline stem cell

Clec11a:

Type C lectin domain family 11 member a

RT-PCR:

Reverse-transcriptionpolymerase chain reaction

qRT-PCR:

Quantitative real-time polymerase chain reaction

AKT:

AKT serine/threonine kinase

CCK8:

Cell Counting Kit-8

EdU:

5-Ethynyl-2′-deoxyuridine

Muc1:

Mucin1

Gpx3:

Glutathione peroxidase 3

Tet1:

Tet methylcytosine dioxygenase 1

References

  1. Zou, K., Yuan, Z., Yang, Z., et al. (2009). Production of offspring from a germline stem cell line derived from neonatal ovaries. Nature Cell Biology, 11(5), 631–636.

    Article  CAS  PubMed  Google Scholar 

  2. Wu, C., Xu, B., Li, X., et al. (2017). Tracing and characterizing the development of transplanted female germline stem cells in vivo. Molecular Therapy, 25(6), 1408–1419.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. White, Y. A. R., Woods, D. C., Takai, Y., et al. (2012). Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women. Nature Medicine, 18(3), 413–421.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Hu, Y., Bai, Y., Chu, Z., et al. (2012). GSK3 inhibitor-BIO regulates proliferation of female germline stem cells from the postnatal mouse ovary. Cell Proliferation, 45(4), 287–298.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Zhu, X., Tian, G. G., Yu, B., et al. (2018). Effects of bisphenol a on ovarian follicular development and female germline stem cells. Archives of Toxicology, 92(4), 1581–1591.

    Article  CAS  PubMed  Google Scholar 

  6. Li, X., Hu, X., Tian, G. G., et al. (2019). C89 induces autophagy of female germline stem cells via inhibition of the PI3K-Akt pathway in vitro. Cells, 8(6), 606.

    Article  CAS  PubMed Central  Google Scholar 

  7. Yuan, X., Tian, G. G., Pei, X., et al. (2021). Spermidine induces cytoprotective autophagy of female germline stem cells in vitro and ameliorates aging caused by oxidative stress through upregulated sequestosome-1/P62 expression. Cell & Bioscience, 11(1), 107.

    Article  CAS  Google Scholar 

  8. Li, B., Hu, X., Yang, Y., et al. (2019) GAS5/miR-21 Axis as a Potential Target to Rescue ZCL-082-Induced Autophagy of Female Germline Stem Cells In Vitro. Molecular Therapy - Nucleic Acids. https://doi.org/10.1016/j.omtn.2019.06.012 

  9. Chen, P., Zhao, X., Tian, G. G., et al. (2021). C28 induced autophagy of female germline stem cells in vitro with changes of H3K27 acetylation and transcriptomics. Gene. https://doi.org/10.1016/j.gene.2020.145150

  10. Jiang, Y., Zhu, D., Liu, W., et al. (2019). Hedgehog pathway inhibition causes primary follicle atresia and decreases female germline stem cell proliferation capacity or stemness. Stem Cell Research & Therapy, 10(1), 198.

    Article  Google Scholar 

  11. Zhang, X., Wei, R., Sun, Y., et al. (2019). AKT3 is a pivotal molecule of Cadherin-22 and GDNF family Receptor-Alpha1 signal pathways regulating self-renewal in female germline stem cells. Stem Cells, 37(8), 1095–1107.

    Article  CAS  PubMed  Google Scholar 

  12. Dwiecki, K., Neunert, G., Polewski, P., et al. (2009). Antioxidant activity of daidzein, a natural antioxidant, and its spectroscopic properties in organic solvents and phosphatidylcholine liposomes. Journal of Photochemistry and Photobiology B: Biology, 96(3), 242–248.

    Article  CAS  Google Scholar 

  13. García-Lafuente, A., Guillamón, E., Villares, A., et al. (2009). Flavonoids as anti-inflammatory agents: implications in cancer and cardiovascular disease. Inflammation research, 58(9), 537–552.

    Article  PubMed  Google Scholar 

  14. Sirotkin, A. V., Alwasel, S. H., & Harrath, A. H. (2021). The influence of plant isoflavones daidzein and equol on female reproductive processes. Pharmaceuticals, 14(4), 373.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Sirotkin, A., Záhoranska, Z., Tarko, A., et al. (2020). Plant isoflavones can prevent adverse effects of benzene on porcine ovarian activity: An in vitro study. Environmental Science and Pollution Research, 27(23), 29589–29598.

    Article  CAS  PubMed  Google Scholar 

  16. Liu, H., Zhang, C., & Zeng, W. (2006). Estrogenic and antioxidant effects of a phytoestrogen daidzein on ovarian germ cells in embryonic chickens. Domestic Animal Endocrinology, 31(3), 258–268.

    Article  PubMed  Google Scholar 

  17. Mlynarczuk, J., Wrobel, M. H., & Kotwica, J. (2011). The adverse effect of phytoestrogens on the synthesis and secretion of ovarian oxytocin in cattle. Reproduction in Domestic Animals, 46(1), 21–28.

    Article  CAS  PubMed  Google Scholar 

  18. Medigović, I. M., Živanović, J. B., Ajdžanović, V. Z., et al. (2015). Effects of soy phytoestrogens on pituitary-ovarian function in middle-aged female rats. Endocrine, 50(3), 764–776.

    Article  PubMed  Google Scholar 

  19. Talsness, C., Grote, K., Kuriyama, S., et al. (2015). Prenatal exposure to the phytoestrogen daidzein resulted in persistent changes in ovarian surface epithelial cell height, folliculogenesis, and estrus phase length in adult sprague-dawley rat offspring. Journal of Toxicology and Environmental Health, Part A, 78(10), 635–644.

    Article  CAS  Google Scholar 

  20. Wang, W., Sun, Y., Liu, J., et al. (2014). Soy isoflavones administered to rats from weaning until sexual maturity affect ovarian follicle development by inducing apoptosis. Food and Chemical Toxicology, 72, 51–60.

    Article  CAS  PubMed  Google Scholar 

  21. Somjen, D., Katzburg, S., Nevo, N., et al. (2008). A daidzein-daunomycin conjugate improves the therapeutic response in an animal model of ovarian carcinoma. The Journal of Steroid Biochemistry and Molecular Biology, 110(1–2), 144–149.

    Article  CAS  PubMed  Google Scholar 

  22. Green, J. M., Alvero, A. B., Kohen, F., et al. (2009). 7-(O)-Carboxymethyl daidzein conjugated to N-t-Boc-hexylenediamine: A novel compound capable of inducing cell death in epithelial ovarian cancer stem cells. Cancer Biology & Therapy, 8(18), 1747–1753.

    Article  CAS  Google Scholar 

  23. Hiraoka, A., Yano, K., Kagami, N., et al. (2001). Stem cell growth factor: In situ hybridization analysis on the gene expression, molecular characterization and in vitro proliferative activity of a recombinant preparation on primitive hematopoietic progenitor cells. The Hematology Journal, 2(5), 307–315.

    Article  CAS  PubMed  Google Scholar 

  24. Shen, B., Vardy, K., Hughes, P., et al. (2019). Integrin Alpha11 is an osteolectin receptor and is required for the maintenance of adult skeletal bone mass. eLife, 8, e42274.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Hiraoka, A., Sugimura, A., Seki, T., et al. (1997). Cloning, expression, and characterization of a cDNA encoding a novel human growth factor for primitive hematopoietic progenitor cells. Proceedings of the National Academy of Sciences - PNAS, 94(14), 7577–7582.

    Article  CAS  Google Scholar 

  26. Shi, R., Hu, J., Li, W., et al. (2019). Protective effects of Clec11a in islets against lipotoxicity via modulation of proliferation and lipid metabolism in mice. Experimental Cell Research, 384(1), 111613.

    Article  CAS  PubMed  Google Scholar 

  27. Brunet, A., Bonni, A., Zigmond, M. J., et al. (1999). Akt promotes cell survival by phosphorylating and inhibiting a forkhead transcription factor. Cell, 96(6), 857–868.

    Article  CAS  PubMed  Google Scholar 

  28. Lawlor, M. A., & Alessi, D. R. (2001). PKB/Akt: A key mediator of cell proliferation, survival and insulin responses? Journal of Cell Science, 114(Pt 16), 2903–2910.

    Article  CAS  PubMed  Google Scholar 

  29. Jin, X., Sun, J., Yu, B., et al. (2017). Daidzein stimulates osteogenesis facilitating proliferation, differentiation, and antiapoptosis in human osteoblast-like MG-63 cells via estrogen receptor-dependent MEK/ERK and PI3K/Akt activation. Nutrition Research, 42, 20–30.

    Article  CAS  PubMed  Google Scholar 

  30. Hiraoka, A. (2008). Leukemia Cell Lines Require Self-Secreted Stem Cell Growth Factor (SCGF) for their proliferation. Leukemia Research, 32(10), 1623–1625.

    Article  CAS  PubMed  Google Scholar 

  31. Stroopinsky, D., Kufe, D., & Avigan, D. (2016). MUC1 in hematological malignancies. Leukaemia & Lymphoma, 57(11), 2489–2498.

    Article  CAS  Google Scholar 

  32. Jalali, S., Shi, J., Buko, A., et al. (2020). Increased glutathione utilization augments tumor cell proliferation in waldenstrom macroglobulinemia. Redox Biology, 36, 101657.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Ito, S., D’Alessio, A. C., Taranova, O. V., et al. (2010). Role of Tet proteins in 5mC to 5hmC conversion, ES-cell self-renewal and inner cell mass specification. Nature, 466(7310), 1129–1133.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Zhang, X., Yang, Y., Xia, Q., et al. (2018). Cadherin 22 participates in the self-renewal of mouse female germ line stem cells via interaction with JAK2 and Β-Catenin. Cellular and Molecular Life Sciences, 75(7), 1241–1253.

    Article  CAS  PubMed  Google Scholar 

  35. Alshehri, M. M., Sharifi-Rad, J., Herrera-Bravo, J., et al. (2021). Therapeutic potential of isoflavones with an emphasis on daidzein. Oxidative Medicine and Cellular Longevity, 2021, 1–15.

    Article  Google Scholar 

  36. Chan, K. K. L., Siu, M. K. Y., Jiang, Y., et al. (2018). Estrogen receptor modulators genistein, daidzein and ERB-041 inhibit cell migration, invasion, proliferation and sphere formation via modulation of FAK and PI3K/AKT signaling in ovarian cancer. Cancer Cell International, 18(1), 65.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Tang, X., Zhang, C., Zeng, W., et al. (2006). Proliferating effects of the flavonoids daidzein and quercetin on cultured chicken primordial germ cells through antioxidant action. Cell Biology International, 30(5), 445–451.

    Article  PubMed  Google Scholar 

  38. Liu, H. Y., & Zhang, C. Q. (2008). Effects of daidzein on messenger ribonucleic acid expression of gonadotropin receptors in chicken ovarian follicles. Poultry Science, 87(3), 541–545.

    Article  CAS  PubMed  Google Scholar 

  39. Nynca, A., Slonina, D., Jablonska, O., et al. (2013). Daidzein affects steroidogenesis and oestrogen receptor expression in medium ovarian follicles of pigs. Acta Veterinaria Hungarica, 61(1), 85–98.

    Article  CAS  PubMed  Google Scholar 

  40. Hu, B., Yu, B., Tang, D., et al. (2016). Daidzein promotes osteoblast proliferation and differentiation in OCT1 cells through stimulating the activation of BMP-2/Smads pathway. Genetics and Molecular Research, 15(2). https://doi.org/10.4238/gmr.15028792

  41. Yu, B., Tang, D., Li, S., et al. (2017). Daidzein promotes proliferation and differentiation in osteoblastic OCT1 cells via activation of the BMP-2/Smads pathway. Die Pharmazie, 72(1), 35.

    CAS  PubMed  Google Scholar 

  42. Le Belle, J. E., Orozco, N. M., Paucar, A. A., et al. (2011). Proliferative neural stem cells have high endogenous ROS levels that regulate self-renewal and neurogenesis in a PI3K/Akt-dependant manner. Cell Stem Cell, 8(1), 59–71.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Godoy-Parejo, C., Deng, C., Liu, W., et al. (2019). Insulin stimulates PI3K/AKT and cell adhesion to promote the survival of individualized human embryonic stem cells. Stem Cells, 37(8), 1030–1041.

    Article  CAS  PubMed  Google Scholar 

  44. Rivera-Gonzalez, G. C., Shook, B. A., Andrae, J., et al. (2016). Skin adipocyte stem cell self-renewal is regulated by a PDGFA/AKT-Signaling axis. Cell Stem Cell, 19(6), 738–751.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Xie, K., Li, Y., Chen, D., et al. (2020). Daidzein supplementation enhances embryo survival by improving hormones, antioxidant capacity, and metabolic profiles of amniotic fluid in sows. Food & Function, 11(12), 10588–10600.

    Article  CAS  Google Scholar 

  46. Yu, Z., Yang, L., Deng, S., et al. (2020). Daidzein ameliorates LPS-induced hepatocyte injury by inhibiting inflammation and oxidative stress. European Journal of Pharmacology, 885, 173399.

    Article  CAS  PubMed  Google Scholar 

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Funding

This study was supported by National Nature Science Foundation of China (81720108017), the National Major Scientific Instruments and Equipment Development Project, National Natural Science Foundation of China (61827814).

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Authors and Affiliations

  1. Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, No. 800 Dongchuan Road, Minhang District, Shanghai, 200240, China

    Fangfang Li, Xiaopeng Hu & Ji Wu

  2. Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan, 750004, China

    Ji Wu

Authors
  1. Fangfang Li
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  2. Xiaopeng Hu
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  3. Ji Wu
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F.L. conducted all the major experiments and wrote the manuscript. J.W. and X.H. conceived the experiments and revised the manuscript.

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Correspondence to Xiaopeng Hu or Ji Wu.

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Li, F., Hu, X. & Wu, J. Daidzein Activates Akt Pathway to Promote the Proliferation of Female Germline Stem Cells through Upregulating Clec11a. Stem Cell Rev and Rep 18, 3021–3032 (2022). https://doi.org/10.1007/s12015-022-10394-0

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