Skip to main content
SpringerLink
Log in

Effects of WT1 gene downregulation on apoptosis in porcine fetal fibroblasts

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Wilms’ tumor gene 1 (WT1) is located on chromosome 11p13. Besides a role in the development of Wilms’ tumor, specific mutations in the Zn finger region are found in Denys-Drash syndrome and Frasier syndrome, both characterized by urogenital abnormalities, sometimes in combination with Wilms’ tumor. Our past study shows that WT1 is expressed in porcine kidney fibroblasts (PKFs) and swine testis cells (ST cells) and is essential for the maintenance of the development and survival of PKFs and ST cells. But we do not know whether WT1 gene was expressed in porcine fetal fibroblasts or not. To further explore whether WT1 was expressed in porcine fetal fibroblasts (PFFs) and its contribution to cell apoptosis, RT-PCR, immunocytochemical staining, and Western blot were used to detect the expression of WT1, the recombinant plasmids of pLV3-WT1 short hairpin ribonucleic acid (shRNA) were used to downregulate the WT1 gene in porcine fetal fibroblasts, and the role of WT1 in cell proliferation was examined by apoptosis analysis also. Our results indicated that WT1 was expressed in PFFs, the pLV3-WT1 shRNA dramatically reduced the expression of WT1, and downregulation of WT1 directly led to early cell apoptosis by downregulating the expression of antiapoptotic gene Bcl-2 and upregulating the expression of proapoptotic gene Bax in PFFs. Our results demonstrate that WT1 is also essential for the maintenance of the survival of PFFs.

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.

Figure 1.
Figure 2.
Figure 3.
Figure 4.

Similar content being viewed by others

References

  • An P, Yin Y, Fan A, Tan W, Gao F, Ma K, Tang B, Zhang X, Li Z (2013) The effects of the WT1 gene on apoptosis and development-related gene expression in porcine kidney fibroblasts and swine testis cells. Mol Reprod Dev 80:414–425

    Article  CAS  PubMed  Google Scholar 

  • Armstrong JF, Pritchard-Jones K, Bickmore WA, Hastie ND, Bard JB (1993) The expression of the Wilms’ tumour gene, WT1, in the developing mammalian embryo. Mech Dev 40:85–97

    Article  CAS  PubMed  Google Scholar 

  • Englert C, Hou X, Maheswaran S, Bennett P, Ngwu C, Re GG, Garvin AJ, Rosner MR, Haber DA (1995) WT1 suppresses synthesis of the epidermal growth factor receptor and induces apoptosis. EMBO J 14:4662–4675

    CAS  PubMed Central  PubMed  Google Scholar 

  • Hohenstein P, Hastie ND (2006) The many facets of the Wilms’ tumour gene, WT1. Hum Mol Genet 15(Spec No 2):R196–R201

    Article  CAS  PubMed  Google Scholar 

  • Hosen N, Shirakata T, Nishida S, Yanagihara M, Tsuboi A, Kawakami M, Oji Y, Oka Y, Okabe M, Tan B et al (2007) The Wilms’ tumor gene WT1-GFP knock-in mouse reveals the dynamic regulation of WT1 expression in normal and leukemic hematopoiesis. Leukemia 21:1783–1791

    Article  CAS  PubMed  Google Scholar 

  • Kijima N, Hosen N, Kagawa N, Hashimoto N, Kinoshita M, Oji Y, Sugiyama H, Yoshimine T (2014) Wilms’ tumor 1 is involved in tumorigenicity of glioblastoma by regulating cell proliferation and apoptosis. Anticancer Res 34:61–67

    CAS  PubMed  Google Scholar 

  • Kreidberg JA, Sariola H, Loring JM, Maeda M, Pelletier J, Housman D, Jaenisch R (1993) WT-1 is required for early kidney development. Cell 74:679–691

    Article  CAS  PubMed  Google Scholar 

  • Kudoh T, Ishidate T, Nakamura T, Toyoshima K, Akiyama T (1996) Constitutive expression of the Wilms tumor suppressor gene WT1 in F9 embryonal carcinoma cells induces apoptotic cell death in response to retinoic acid. Oncogene 13:1431–1439

    CAS  PubMed  Google Scholar 

  • Lai L, Prather RS (2003) Production of cloned pigs by using somatic cells as donors. Cloning Stem Cells 5:233–241

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Niksic M, Slight J, Sanford JR, Caceres JF, Hastie ND (2004) The Wilms’ tumour protein (WT1) shuttles between nucleus and cytoplasm and is present in functional polysomes. Hum Mol Genet 13:463–471

    Article  CAS  PubMed  Google Scholar 

  • Park S, Schalling M, Bernard A, Maheswaran S, Shipley GC, Roberts D, Fletcher J, Shipman R, Rheinwald J, Demetri G et al (1993) The Wilms tumour gene WT1 is expressed in murine mesoderm-derived tissues and mutated in a human mesothelioma. Nat Genet 4:415–420

    Article  CAS  PubMed  Google Scholar 

  • Pelletier J, Schalling M, Buckler AJ, Rogers A, Haber DA, Housman D (1991) Expression of the Wilms’ tumor gene WT1 in the murine urogenital system. Genes Dev 5:1345–1356

    Article  CAS  PubMed  Google Scholar 

  • Pritchard-Jones K, Fleming S, Davidson D, Bickmore W, Porteous D, Gosden C, Bard J, Buckler A, Pelletier J, Housman D et al (1990) The candidate Wilms’ tumour gene is involved in genitourinary development. Nature 346:194–197

    Article  CAS  PubMed  Google Scholar 

  • Rao MK, Pham J, Imam JS, MacLean JA, Murali D, Furuta Y, Sinha-Hikim AP, Wilkinson MF (2006) Tissue-specific RNAi reveals that WT1 expression in nurse cells controls germ cell survival and spermatogenesis. Genes Dev 20:147–152

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Reddy JC, Licht JD (1996) The WT1 Wilms’ tumor suppressor gene: how much do we really know. Biochim Biophys Acta 1287:1–28

    PubMed  Google Scholar 

  • Rivera MN, Haber DA (2005) Wilms’ tumour: connecting tumorigenesis and organ development in the kidney. Nat Rev Cancer 5:699–712

    Article  CAS  PubMed  Google Scholar 

  • Scholz H, Kirschner KM (2005) A role for the Wilms’ tumor protein WT1 in organ development. Physiology (Bethesda) 20:54–59

    Article  CAS  Google Scholar 

  • Schreiner CL, Jones EE (1987) Metabolism of methionine and methionine hydroxy analogue by porcine kidney fibroblasts. J Nutr 117:1541–1549

    CAS  PubMed  Google Scholar 

  • Siegel JF, Delakas D, Rai S, Kushner L (1996) Unilateral nephrectomy induces the expression of the Wilms tumor gene in the contralateral kidney of the adult rat. J Urol 156:688–692

    Article  CAS  PubMed  Google Scholar 

  • Tatsumi N, Oji Y, Tsuji N, Tsuda A, Higashio M, Aoyagi S, Fukuda I, Ito K, Nakamura J, Takashima S et al (2008) Wilms’ tumor gene WT1-shRNA as a potent apoptosis-inducing agent for solid tumors. Int J Oncol 32:701–711

    CAS  PubMed  Google Scholar 

  • Uesugi K, Hiasa Y, Tokumoto Y, Mashiba T, Koizumi Y, Hirooka M, Abe M, Matsuura B, Onji M (2013) Wilms’ tumor 1 gene modulates Fas-related death signals and anti-apoptotic functions in hepatocellular carcinoma. J Gastroenterol 48:1069–1080

    Article  CAS  PubMed  Google Scholar 

  • Walker C, Rutten F, Yuan X, Pass H, Mew DM, Everitt J (1994) Wilms’ tumor suppressor gene expression in rat and human mesothelioma. Cancer Res 54:3101–3106

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation (No. 31072027), National Basic Research Program (No. 2011CB944204), Postdoctoral Special Funded Projects (No. 201003536), and Innovative Research Team in University (IRT1248) in China.

Author information

Authors and Affiliations

  1. Jilin Provincial Key Laboratory of Animal Embryo Engineering, The Center for Animal Embryo Engineering of Jilin Province, College of Veterinary Medicine, Jilin University, 5333 Xi An Da Lu, Changchun, 130062, Jilin, China

    Peipei An, Aibing Wang, Wentao Tan, Fei Gao, Anran Fan, Bo Tang, Xueming Zhang & Ziyi Li

  2. State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences of CAAS, Chinese Academy of Agricultural Science, Changchun, 130112, Jilin, China

    Peipei An

  3. College of Animal Science, Jilin University, Changchun, 130062, Jilin, China

    Yu Ding

Authors
  1. Peipei An
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yu Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Aibing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wentao Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fei Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anran Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bo Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xueming Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ziyi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Xueming Zhang or Ziyi Li.

Additional information

Editor: T. Okamoto

Peipei An and Yu Ding contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

An, P., Ding, Y., Wang, A. et al. Effects of WT1 gene downregulation on apoptosis in porcine fetal fibroblasts. In Vitro Cell.Dev.Biol.-Animal 50, 555–561 (2014). https://doi.org/10.1007/s11626-014-9738-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-014-9738-1

Keywords

Search

Navigation