Skip to main content

Advertisement

SpringerLink
Log in

miR-3074-5p Promotes the Apoptosis but Inhibits the Invasiveness of Human Extravillous Trophoblast-Derived HTR8/SVneo Cells In Vitro

  • Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

Objective

The objective of this study was to observe the effects of the overexpression of miR-3074-5p in human trophoblast cells in vitro.

Design

Experimental in vitro study in HTR8/SVneo cells.

Methods

HTR8/SVneo cells were transfected with miR-3074-5p mimic. The cell apoptosis and invasion were measured via flow cytometry and transwell assay, respectively. The expression levels of P53, Cyclin Dependent Kinase Inhibitor I B (P27), BCL-2, BCL2 associated X (BAX), and BCL2 like 14 (BCL-G) in HTR8/ SVneo cells were determined by Western blot. The alterations in gene expression profile of HTR8/SVneo cells were evaluated by complementary DNA microarray assay, and the differential expressions of dihydrolipoamide S-succinyltransferase (DLST), growth-associated protein 43 (GAP43), runt-related transcription factor 2 (RUNX2), and C-C type chemokine receptor 3 (CCR3) were validated by Western blot. Biofunctions of these differentially expressed genes were enriched by Gene Ontology analysis.

Results

The overexpression of miR-3074-5p in HTR8/SVneo cells promoted cell apoptosis but inhibited cell invasion, being accompanied by the significantly elevated expressions of P27, BCL-2, and BCL-G. Meanwhile, an increased expression of P27 and P57 was also detected in a small sample size of placental villi of recurrent miscarriage (RM) patients. Totally, 41 I genes and 397 genes were screened out, respectively, to be downregulated or upregulated at least by 2-folds in miR-3074-5p overexpressed HTR8/SVneo cells. These differentially expressed genes were involved in several important functions related to pregnancy. Subsequently, the reduced expressions of DLST and GAP43 proteins, as well as the increased expressions of CCR3 and RUNX2 proteins, were validated in miR-3074-5p overexpressed HTR8/SVneo cells. Conclusion: These data suggested a potential contribution of miR-3074-5p in the pathogenesis of RM by disturbing the normal activities of trophoblast cells.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Brezina PR, Kutteh WH. Classic and cutting-edge strategies for the management of early pregnancy loss. Obstet Gynecol Clin North Am. 2014;41(1):1–18.

    PubMed  Google Scholar 

  2. Rai R, Regan L. Recurrent miscarriage. Lancet. 2006;368(9535): 601–611.

    PubMed  Google Scholar 

  3. Lunghi L, Ferretti ME, Medici S, et al. Control of human tropho-blast function. Reprod Biol Endocrinol. 2007;5(1):6.

    PubMed  PubMed Central  Google Scholar 

  4. Winship A, Correia J, Krishnan T, et al. Blocking endogenous leukemia inhibitory factor during placental development in mice leads to abnormal placentation and pregnancy loss. Sci Rep. 2015; 5(1):13237.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Wightman B, Ha I, Ruvkun G. Posttranscriptional regulation of the heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans. Cell. 1993;75(5):855–862.

    CAS  PubMed  Google Scholar 

  6. Battel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116(2):281–297.

    Google Scholar 

  7. Harun R, Ruban L, Matin M, et al. Cytotrophoblast stem cell lines derived from human embryonic stem cells and their capacity to mimic invasive implantation events. Hum Reprod. 2006;21(6): 1349–1358.

    CAS  PubMed  Google Scholar 

  8. Galliano D, Pellicer A. MicroRNA and implantation. Fertil Steril. 2014;101(6):1531–1544.

    CAS  Google Scholar 

  9. Hull ML, Nisenblat V. Tissue and circulating microRNA influence reproductive function in endometrial disease. Reprod Biomed Online. 2013;27(5):515–529.

    PubMed  Google Scholar 

  10. Dong F, Zhang Y, Xia F, et al. Genome-wide miRNA profiling of villus and decidua of recurrent spontaneous abortion patients. Reproduction. 2014;148(1):33–41.

    CAS  PubMed  Google Scholar 

  11. Qin W, Tang Y, Yang N, et al. Potential role of circulating micro-RNAs as a biomarker for unexplained recurrent spontaneous abortion. Fertil Steril. 2016;105(5):1247–1254 e1243.

    CAS  PubMed  Google Scholar 

  12. Tang L, Gao C, Gao L, et al. Expression profile of micro-RNAs and functional annotation analysis of their targets in human chorionic villi from early recurrent miscarriage. Gene. 2016;576(1 pt 2):366–371.

    CAS  PubMed  Google Scholar 

  13. Gu Y, Zhang X, Yang Q, et al. Aberrant placental villus expression of miR-486-3p and miR-3074-5p in recurrent miscarriage patients and uterine expression of these micrornas during early pregnancy in mice. Gynecol Obstet Invest. 2015;1(1):1.

    CAS  Google Scholar 

  14. Abou-Kheir W, Barrak J, Hadadeh O, et al. HTR-8/SVneo cell line contains a mixed population of cells. Placenta. 2017; 50(1):1–7.

    CAS  PubMed  Google Scholar 

  15. Yang M, Chen Y, Chen L, et al. miR-15b-AG02 play a critical role in HTR8/SVneo invasion and in a model of angiogenesis defects related to inflammation. Placenta. 2016;41(1):62–73.

    CAS  PubMed  Google Scholar 

  16. Ding GC, Chen M, Wang YX, et al. MicroRNA-128a-induced apoptosis in HTR-8/SVneo trophoblast cells contributes to preeclampsia. Biomed Pharmacother. 2016;81(1):63–70.

    CAS  Google Scholar 

  17. Graham CH, Hawley TS, Hawley RG, et al. Establishment and characterization of first trimester human trophoblast cells with extended lifespan. Exp Cell Res. 1993;206(2):204–211.

    CAS  PubMed  Google Scholar 

  18. Salic A, Mitchison TJ. A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci USA. 2008;105(7):2415–2420.

    CAS  PubMed  Google Scholar 

  19. Szklarczyk D, Franceschini A, Wyder S, et al. STRING vlO: protein-protein interaction networks, integrated over the tree of life. Nucleic Acids Res. 2015;43(database issue):D447–D452.

    CAS  PubMed  Google Scholar 

  20. Unek G, Ozmen A, Kipmen-Korgun D, et al. Immunolocalization of PCNA, Ki67, p27 and p57 in normal and dexamethasone-induced intrauterine growth restriction placental development in rat. Acta Histochemica. 2012;114(1):31–40.

    CAS  PubMed  Google Scholar 

  21. Dior UP, Kogan L, Chill HH, et al. Emerging roles of microRNA in the embryo-endometrium cross talk. Semin Reprod Med. 2014; 32(5):402–409.

    CAS  PubMed  Google Scholar 

  22. Ge YZ, Xu LW, Xu Z, et al. Expression profiles and clinical significance of MicroRNAs in papillary renal cell carcinoma: a STROBE-compliant observational study. Medicine. 2015;94(16): e767.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Korkmaz C, Sakinci M, Akyol SN, et al. Location of proliferating cell nuclear antigen and p53 protein in human first trimester and term placenta. Anal Quant Cytopathol Histpathol. 2013;35(6): 335–343.

    PubMed  Google Scholar 

  24. Nadeem L, Brkic J, Chen YF, et al. Cytoplasmic mislocalization of p27 and CDK2 mediates the anti-migratory and antiproliferative effects of Nodal in human trophoblast cells. J Cell Sci. 2013;126(pt 2):445–453.

    CAS  PubMed  Google Scholar 

  25. Gu Y, He Y, Zhang X, et al. Deficiency of monoclonal non-specific suppressor factor beta (MNSFB) promotes pregnancy loss in mice. Mol Reprod Dev. 2015;82(6):475–488.

    CAS  PubMed  Google Scholar 

  26. Suzuki HI, Young RA, Sharp PA. Super-enhancer-mediated RNA processing revealed by integrative MicroRNA network analysis. Cell. 2017;168(6):1000–1014 el015.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Borriello A, Bencivenga D, Criscuolo M, et al. Targeting p27Kipl protein: its relevance in the therapy of human cancer. Expert Opin Ther Targets. 2011;15(6):677–693.

    CAS  PubMed  Google Scholar 

  28. Podmirseg SR, Jakel H, Ranches GD, et al. Caspases uncouple p27(Kipl) from cell cycle regulated degradation and abolish its ability to stimulate cell migration and invasion. Oncogene. 2016; 35(35):4580–4590.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Wei P, Jin X, Zhang XS, et al. Expression of Bcl-2 and p53 at the fetal-maternal interface of rhesus monkey. Reprod Biol Endocrinol. 2005;3(1):4.

    PubMed  PubMed Central  Google Scholar 

  30. Gross A. BCL-2 family proteins as regulators of mitochondria metabolism. Biochim Biophys Acta. 2016;1857(8):1243–1246.

    CAS  PubMed  Google Scholar 

  31. Bai Y, Wang J, Han J, et al. BCL2L10 inhibits growth and metastasis of hepatocellular carcinoma both in vitro and in vivo. Mol Carcinog. 2016;1(1):1.

    Google Scholar 

  32. Unek G, Ozmen A, Mendilcioglu I, et al. The expression of cell cycle related proteins PCNA, Ki67, p27 and p57 in normal and preeclamptic human placentas. Tissue & cell. 2014;46(3): 198–205.

    CAS  Google Scholar 

  33. Gundogan F, Elwood G, Mark P, et al. Ethanol-induced oxidative stress and mitochondrial dysfunction in rat placenta: relevance to pregnancy loss. Alcohol Clin Exp Res. 2010;34(3):415–423.

    CAS  PubMed  Google Scholar 

  34. Wang JM, Gu Y, Zhang Y, et al. Deep-sequencing identification of differentially expressed miRNAs in decidua and villus of recurrent miscarriage patients. Arch Gynecol Obstet. 2016;293(5): 1125–1135.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Miled C, Pontoglio M, Garbay S, et al. A genomic map of p53 binding sites identifies novel p53 targets involved in an apoptotic network. Cancer Res. 2005;65(12):5096–5104.

    CAS  PubMed  Google Scholar 

  36. Zhang YY, Li X, Qian SW, et al. Down-regulation of type I Runx2 mediated by dexamethasone is required for 3T3-L1 adi-pogenesis. Mol Endocrinol. 2012;26(5):798–808.

    PubMed  PubMed Central  Google Scholar 

  37. Henriquez B, Hepp M, Merino P, et al. C/EBPbeta binds the P1 promoter of the Runx2 gene and up-regulates Runx2 transcription in osteoblastic cells. J Cell Physiol. 2011;226(11):3043–3052.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Anderson NM, Li D, Peng HL, et al. The TCA cycle transferase DLST is important for MYC-mediated leukemogenesis. Leukemia. 2016;30(6): 1365–1374.

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Diaz-Munoz MD, Bell SE, Fairfax K, et al. The RNA-binding protein HuR is essential for the B cell antibody response. Nat Immunol. 2015;16(4):415–425.

    CAS  Google Scholar 

  40. Zhang H, Wu F, Kong X, et al. Nerve growth factor improves functional recovery by inhibiting endoplasmic reticulum stress-induced neuronal apoptosis in rats with spinal cord injury. J Transl Med. 2014;12(1):130.

    PubMed  PubMed Central  Google Scholar 

  41. Luo Y, Shen H, Liu HS, et al. CART peptide induces neurore-generation in stroke rats. J Cereb Blood Flow Metab. 2013;33(2): 300–310.

    CAS  PubMed  Google Scholar 

  42. Hannan NJ, Jones RL, White CA, et al. The chemokines, CX3CL1, CCL14, and CCL4, promote human trophoblast migration at the fetomaternal interface. Biol Reprod. 2006;74(5):896–904.

    CAS  PubMed  Google Scholar 

  43. Athanassakis I, Papadimitriou L, Vassiliadis S. Murine ectopla-cental cone-derived trophoblast cells express chemokine receptors. J Reprod Immunol. 2001;50(2):105–119.

    CAS  PubMed  Google Scholar 

  44. Chang MC, Tsai YL, Liou EJ, et al. Effect of butyrate on collagen expression, cell viability, cell cycle progression and related pro-teins expression of MG-63 osteoblastic cells. PloS one. 2016; 11(11):e0165438.

    PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Second Hospital of Tianjin Medical University, Tianjin, China

    Yan Gu PhD & Jian-Mei Wang MD

  2. Key Laboratory of Reproduction Regulation of NHFPC, Shanghai Institute of Planned Parenthood Research, Fudan University, 2140 Xie Tu Road, Shanghai, China

    Yan Shi MS, Wen-Wen Gu PhD, Ya-Ping He PhD, Hua-Jun Zheng PhD, Xuan Zhang PhD & Jian Wang PhD

  3. School of Pharmacy, Fudan University, Shanghai, China

    Qian Yang PhD & Jian Wang PhD

Authors
  1. Yan Gu PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yan Shi MS
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qian Yang PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wen-Wen Gu PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ya-Ping He PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hua-Jun Zheng PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xuan Zhang PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jian-Mei Wang MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jian Wang PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Xuan Zhang PhD or Jian Wang PhD.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gu, Y., Shi, Y., Yang, Q. et al. miR-3074-5p Promotes the Apoptosis but Inhibits the Invasiveness of Human Extravillous Trophoblast-Derived HTR8/SVneo Cells In Vitro. Reprod. Sci. 25, 690–699 (2018). https://doi.org/10.1177/1933719117725823

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1177/1933719117725823

Keywords

Search

Navigation