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Silencing of circ_0007299 suppresses proliferation, migration, and invasiveness and promotes apoptosis of ectopic endometrial stromal cells in endometriosis via miR-424-5p-dependent modulation of CREB1

  • General Gynecology
  • Published:
Archives of Gynecology and Obstetrics Aims and scope Submit manuscript

Abstract

Background

The abnormality of endometrial stromal cells (ESCs) can contribute to endometriosis pathogenesis. Circular RNAs (circRNAs) possess critical roles in endometriosis pathogenesis. Here, we defined the activity and mechanism of human circ_0007299 in the regulation of ectopic ESCs in vitro.

Methods

Circ_0007299, miR-424-5p and cAMP response element-binding protein 1 (CREB1) were quantified by qRT-PCR or immunoblotting. Cell viability, proliferation, apoptosis, invasion and motility were gauged by CCK-8, 5-Ethynyl-2ʹ-Deoxyuridine (EdU), flow cytometry, transwell, and wound-healing assays, respectively. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were used to verify the direct relationship between miR-424-5p and circ_0007299 or CREB1.

Results

Our data showed that circ_0007299 was upregulated in human ectopic endometrium tissues and ectopic ESCs. Silencing endogenous circ_0007299 impeded the proliferation, invasiveness, and motility and enhanced apoptosis of ectopic ESCs. Mechanistically, circ_0007299 regulated miR-424-5p expression. Moreover, circ_0007299 silencing impeded the proliferation, invasiveness, and motility and enhanced apoptosis of ectopic ESCs via its regulation on miR-424-5p. CREB1 was identified as a direct miR-424-5p target, and miR-424-5p overexpression suppressed ectopic ESC proliferation, migration, and invasiveness and promoted apoptosis by downregulating CREB1. Furthermore, circ_0007299 positively modulated CREB1 expression through miR-424-5p competition.

Conclusion

Our findings establish that circ_0007299 silencing impedes the proliferation, invasiveness, and motility and promotes apoptosis of ectopic ESCs at least in part via miR-424-5p-dependent modulation of CREB1.

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References

  1. Arafah M et al (2021) Endometriosis: a comprehensive review. Adv Anat Pathol 28(1):30–43

    Article  Google Scholar 

  2. Saunders PTK et al (2021) Endometriosis: etiology, pathobiology, and therapeutic prospects. Cell 184(11):2807–2824

    Article  CAS  Google Scholar 

  3. Taylor HS et al (2021) Endometriosis is a chronic systemic disease: clinical challenges and novel innovations. Lancet 397(10276):839–852

    Article  CAS  Google Scholar 

  4. Altayyeb A et al (2020) Characterization of mechanical signature of eutopic endometrial stromal cells of endometriosis patients. Reprod Sci 27(1):364–374

    Article  CAS  Google Scholar 

  5. Lu Q et al (2020) T-cadherin inhibits invasion and migration of endometrial stromal cells in endometriosis. Hum Reprod 35(1):145–156

    Article  CAS  Google Scholar 

  6. McQueen DB et al (2021) Redefining chronic endometritis: the importance of endometrial stromal changes. Fertil Steril 116(3):855–861

    Article  Google Scholar 

  7. Chen L et al (2021) CircRNA in cancer: fundamental mechanism and clinical potential. Cancer Lett 505:49–57

    Article  CAS  Google Scholar 

  8. Das A et al (2021) Identification of potential circRNA-microRNA-mRNA regulatory network in skeletal muscle. Front Mol Biosci 8:762185

    Article  CAS  Google Scholar 

  9. Wu J et al (2021) Construction and topological analysis of an endometriosis-related exosomal circRNA-miRNA-mRNA regulatory network. Aging 13(9):12607–12630

    Article  CAS  Google Scholar 

  10. Tu J et al (2021) Current and future roles of circular RNAs in normal and pathological endometrium. Front Endocrinol 12:668073

    Article  Google Scholar 

  11. Dana PM et al (2020) Circular RNA as a potential diagnostic and/or therapeutic target for endometriosis. Biomark Med 14(13):1277–1287

    Article  CAS  Google Scholar 

  12. Wu R et al (2020) Circular RNA expression profiling and bioinformatic analysis of cumulus cells in endometriosis infertility patients. Epigenomics 12(23):2093–2108

    Article  CAS  Google Scholar 

  13. Li Q et al (2020) Estrogen-decreased hsa_circ_0001649 promotes stromal cell invasion in endometriosis. Reproduction 160(4):511–519

    Article  CAS  Google Scholar 

  14. Xu A et al (2020) Down-regulation of circ_0061140 attenuates ectopic endometrial cell proliferation, migration and invasion in endometriosis via inactivating Notch2. Gene 757:1449

    Article  Google Scholar 

  15. Jiang N et al (2020) Upregulated circular RNA hsa_circ_0008433 regulates pathogenesis in endometriosis via miRNA. Reprod Sci 27(11):2002–2017

    Article  CAS  Google Scholar 

  16. Liu D et al (2021) Knockdown of circ_0075503 suppresses cell migration and invasion by regulating miR-15a-5p and KLF12 in endometriosis. Mol Cell Biochem 476(10):3845–3856

    Article  CAS  Google Scholar 

  17. Dong L et al (2020) Circ_0007331 knock-down suppresses the progression of endometriosis via miR-200c-3p/HiF-1α axis. J Cell Mol Med 24(21):12656–12666

    Article  CAS  Google Scholar 

  18. Sun L et al (2021) Circular RNA PIP5K1A (circPIP5K1A) accelerates endometriosis progression by regulating the miR-153-3p/Thymosin Beta-4 X-Linked (TMSB4X) pathway. Bioengineered 12(1):7104–7118

    Article  Google Scholar 

  19. Feng Y et al (2020) LncRNA MALAT1 inhibits apoptosis of endometrial stromal cells through miR-126-5p-CREB1 axis by activating PI3K-AKT pathway. Mol Cell Biochem 475(1–2):185–194

    Article  CAS  Google Scholar 

  20. Liu S et al (2021) Knockdown of lncRNA H19 suppresses endometriosis in vivo. Braz J Med Biol Res 54(4):e10117

    Article  CAS  Google Scholar 

  21. Deshmukh P et al (2021) The miRISC component AGO2 has multiple binding sites for Nup358 SUMO-interacting motif. Biochem Biophys Res Commun 556:45–52

    Article  CAS  Google Scholar 

  22. Donnelly S et al (2021) Commandeering the mammalian Ago2 miRNA network: a newly discovered mechanism of helminth immunomodulation. Trends Parasitol 37(12):1031–1033

    Article  CAS  Google Scholar 

  23. Guo Y et al (2021) Circ3823 contributes to growth, metastasis and angiogenesis of colorectal cancer: involvement of miR-30c-5p/TCF7 axis. Mol Cancer 20(1):93

    Article  CAS  Google Scholar 

  24. Liu F et al (2021) Hsa_circ_0088212-mediated miR-520 h/APOA1 axis inhibits osteosarcoma progression. Transl Oncol 14(12):101219

    Article  CAS  Google Scholar 

  25. Leitão AL et al (2022) A structural view of miRNA biogenesis and function. Noncoding RNA 8(1):10

    Article  Google Scholar 

  26. Ma LL et al (2021) Tumor suppressor miR-424-5p abrogates ferroptosis in ovarian cancer through targeting ACSL4. Neoplasma 68(1):165–173

    Article  CAS  Google Scholar 

  27. Cui Y et al (2020) miR-424-5p regulates cell proliferation and migration of esophageal squamous cell carcinoma by targeting SIRT4. J Cancer 11(21):6337–6347

    Article  CAS  Google Scholar 

  28. Dai W et al (2020) miR-424-5p promotes the proliferation and metastasis of colorectal cancer by directly targeting SCN4B. Pathol Res Pract 216(1):152731

    Article  CAS  Google Scholar 

  29. Lin M et al (2019) miR-424-5p maybe regulate blood-brain barrier permeability in a model in vitro with Abeta incubated endothelial cells. Biochem Biophys Res Commun 517(3):525–531

    Article  CAS  Google Scholar 

  30. Xiang Y et al (2020) LncRNA MEG3 targeting miR-424-5p via MAPK signaling pathway mediates neuronal apoptosis in ischemic stroke. Aging 12(4):3156–3174

    Article  CAS  Google Scholar 

  31. Yu H et al (2020) Circular RNA circ_0029589 regulates proliferation, migration, invasion, and apoptosis in ox-LDL-stimulated VSMCs by regulating miR-424–5p/IGF2 axis. Vascul Pharmacol 135:106782

    Article  CAS  Google Scholar 

  32. Wang S et al (2021) Effects of CDKN2B-AS1 on cellular proliferation, invasion and AKT3 expression are attenuated by miR-424-5p in a model of ovarian endometriosis. Reprod Biomed Online 42(6):1057–1066

    Article  Google Scholar 

  33. Huan Q et al (2021) LncRNA AFAP1-AS1 regulates proliferation and apoptosis of endometriosis through activating STAT3/TGF-β/Smad signaling via miR-424-5p. J Obstet Gynaecol Res 47(7):2394–2405

    Article  CAS  Google Scholar 

  34. Long Q et al (2020) Early maternal separation accelerates the progression of endometriosis in adult mice. Reprod Biol Endocrinol 18(1):63

    Article  CAS  Google Scholar 

  35. Lv X et al (2018) Analysis of the oncogene BRAF mutation and the correlation of the expression of wild-type BRAF and CREB1 in endometriosis. Int J Mol Med 41(3):1349–1356

    CAS  Google Scholar 

Download references

Funding

The present study was supported by National Natural Science Fund Regional Foundation Project (No. 81760879).

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Authors and Affiliations

  1. Gansu University of Traditional Chinese Medicine, No.35, Dingxi East Road, Chengguan District, Lanzhou City, 730000, Gansu Province, China

    Haiyan Mao, Xiaohua Zhang, Xiujia Ji, Cancan Huang & Quansheng Wu

  2. Traditional Medical Diagnosis and Treatment Center, Gansu Provincial Hospital, No.204, Donggang West Road, Lanzhou City, 730000, Gansu Province, China

    Haiyan Mao

  3. Lanzhou University Second Hospital, Lanzhou City, Gansu Province, China

    Lu Yin

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  1. Haiyan Mao
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  2. Xiaohua Zhang
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  3. Lu Yin
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Corresponding author

Correspondence to Quansheng Wu.

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The authors declare that they have no conflicts of interest.

Ethical approval

All tissue samples were collected with written informed consent in accordance with the Declaration of Helsinki and with the approval of the Ethics Committee of Gansu University of Traditional Chinese Medicine.

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Supplementary Information

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404_2022_6650_MOESM1_ESM.tif

Supplementary file 1: Supplement Figure 1. Representative immunocytochemical image showing vimentin staining in the isolated ectopic ESCs. n = 3 independent biological replicates. (TIF 354 KB)

404_2022_6650_MOESM2_ESM.tif

Supplementary file 2: Supplement Figure 2. Expression of PCNA and Bax proteins by western blot in ectopic ESCs transfected with si-circ_0007299 or si-NC. n = 3 independent biological replicates. (TIF 879 KB)

404_2022_6650_MOESM3_ESM.tif

Supplementary file 3: Supplement Figure 3. Selection of miR-424-5p and CREB1 in this study. (A) The expression of miR-195-5p, miR-15a-5p and miR-424-5p by qRT-PCR in ectopic ESCs transfected with si-NC or si-circ_0007299. (B) The mRNA expression of KLF12, YAP1, AKT3, TGFBR1, NOTCH2 and CREB1 by qRT-PCR in ectopic ESCs transfected with miR-NC mimic or miR-424-5p mimic. N = 3 independent biological replicates. **P < 0.01, ***P < 0.001, ****P < 0.0001. (TIF 410 KB)

404_2022_6650_MOESM4_ESM.tif

Supplementary file 4: Supplement Figure 4. Effects of CREB1 knockdown on cell proliferation, apoptosis, invasiveness, and motility. (A-G) Ectopic ESCs were transiently transfected with si-CREB1 or si-con. (A) Representative immunoblotting revealing CREB1 protein level in ectopic ESCs transfected as indicated. (B) Cell viability by CCK-8 assay. (C) Cell proliferation by EdU assay. (D) Cell apoptosis by flow cytometry. (E) Transwell assay for cell invasiveness ability. (F) Wound-healing assay for cell migration. (G) Representative immunoblotting showing E-cadherin and N-cadherin expression levels in ectopic ESCs transfected as indicated. n = 6 or 3 independent biological replicates. **P < 0.01, ***P < 0.001, ****P < 0.0001. (TIF 1540 KB)

Supplementary file 5: Supplement Table 1. Sequences of qRT-PCR primers. (DOCX 172 KB)

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Mao, H., Zhang, X., Yin, L. et al. Silencing of circ_0007299 suppresses proliferation, migration, and invasiveness and promotes apoptosis of ectopic endometrial stromal cells in endometriosis via miR-424-5p-dependent modulation of CREB1. Arch Gynecol Obstet 307, 149–161 (2023). https://doi.org/10.1007/s00404-022-06650-w

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  • DOI: https://doi.org/10.1007/s00404-022-06650-w

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