Skip to main content

Advertisement

Log in

Down-regulation of Wnt10a by RNA interference inhibits proliferation and promotes apoptosis in mouse embryonic palatal mesenchymal cells through Wnt/β-catenin signaling pathway

  • Original Paper
  • Published:
Journal of Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

Cleft palate is one of the most common birth defects. Both environmental and genetic factors are involved in this disorder. Here, we investigated the function of Wnt10a in proliferation and apoptosis of mouse embryonic palatal mesenchymal (MEPM) cells. Expression of Wnt10a was down-regulated at both the mRNA and protein levels in transfected MEPM cells containing Wnt10a-specific small hairpin RNA (shRNA) plasmid. Down-regulation of Wnt10a inhibited cell proliferation and induced cell cycle arrest in the S phase in MEPM cells. Moreover, apoptosis was significantly increased in MEPM cells of Wnt10a gene silencing. Finally, the expression of β-catenin was markedly reduced in MEPM cells transfected with shRNA plasmid, indicating that the canonical Wnt/β-catenin signaling pathway was involved in the alterations of cell proliferation and apoptosis induced by Wnt10a knockdown. Thus, our findings reveal that Wnt10a regulates proliferation and apoptosis of MEPM cells at least partially through the canonical Wnt/β-catenin signaling pathway.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Beaty TH, Hetmanski JB, Fallin MD, Park JW, Sull JW, McIntosh I, Liang KY, Vanderkolk CA, Redett RJ, Boyadjiev SA et al (2006) Analysis of candidate genes on chromosome 2 in oral cleft case-parent trios from three populations. Hum Genet 120:501–518

    Article  PubMed  CAS  Google Scholar 

  2. Cawthorn WP, Bree AJ, Yao Y, Du B, Hemati N, Martinez-Santibanez G, MacDougald OA (2012) Wnt6, Wnt10a and Wnt10b inhibit adipogenesis and stimulate osteoblastogenesis through a beta-catenin-dependent mechanism. Bone 50:477–489

    Article  PubMed  CAS  Google Scholar 

  3. Chiquet BT, Blanton SH, Burt A, Ma D, Stal S, Mulliken JB, Hecht JT (2008) Variation in WNT genes is associated with non-syndromic cleft lip with or without cleft palate. Hum Mol Genet 17:2212–2218

    Article  PubMed  CAS  Google Scholar 

  4. Christensen K, Juel K, Herskind AM, Murray JC (2004) Long term follow up study of survival associated with cleft lip and palate at birth. BMJ 328:1405

    Article  PubMed  Google Scholar 

  5. Dixon MJ, Marazita ML, Beaty TH, Murray JC (2011) Cleft lip and palate: understanding genetic and environmental influences. Nat Rev Genet 12:167–178

    Article  PubMed  CAS  Google Scholar 

  6. Han J, Xiao Y, Lin J, Li Y (2006) PDGF-C controls proliferation and is down-regulated by retinoic acid in mouse embryonic palatal mesenchymal cells. Birth Defects Res B Dev Reprod Toxicol 77:438–444

    Article  PubMed  CAS  Google Scholar 

  7. He F, Xiong W, Yu X, Espinoza-Lewis R, Liu C, Gu S, Nishita M, Suzuki K, Yamada G, Minami Y, Chen Y (2008) Wnt5a regulates directional cell migration and cell proliferation via Ror2-mediated noncanonical pathway in mammalian palate development. Development 135(23):3871–3879

    Article  PubMed  CAS  Google Scholar 

  8. Hsu RJ, Ho JY, Cha TL, Yu DS, Wu CL, Huang WP, Chu P, Chen YH, Chen JT, Yu CP (2012) WNT10A plays an oncogenic role in renal cell carcinoma by activating WNT/beta-catenin pathway. PLoS One 7:e47649

    Article  PubMed  CAS  Google Scholar 

  9. Hu X, Chen Z, Mao X, Tang S (2011) Effects of phenytoin and Echinacea purpurea extract on proliferation and apoptosis of mouse embryonic palatal mesenchymal cells. J Cell Biochem 112:1311–1317

    Article  PubMed  CAS  Google Scholar 

  10. Jin YR, Han XH, Taketo MM, Yoon JK (2012) Wnt9b-dependent FGF signaling is crucial for outgrowth of the nasal and maxillary processes during upper jaw and lip development. Development 139:1821–1830

    Article  PubMed  CAS  Google Scholar 

  11. Kagalwalla AF, Akhtar N, Woodruff SA, Rea BA, Masterson JC, Mukkada V, Parashette KR, Du J, Fillon S, Protheroe CA, Lee JJ, Amsden K, Melin-Aldana H, Capocelli KE, Furuta GT, Ackerman SJ (2012) Eosinophilic esophagitis: epithelial mesenchymal transition contributes to esophageal remodeling and reverses with treatment. J Allergy Clin Immunol 129(5):1387–1396.e7

    Article  PubMed  Google Scholar 

  12. Kokkinos MI, Wafai R, Wong MK, Newgreen DF, Thompson EW, Waltham M (2007) Vimentin and epithelial–mesenchymal transition in human breast cancer—observations in vitro and in vivo. Cells Tissues Organs 185(1–3):191–203

    Article  PubMed  CAS  Google Scholar 

  13. Lan Y, Ryan RC, Zhang Z, Bullard SA, Bush JO, Maltby KM, Lidral AC, Jiang R (2006) Expression of Wnt9b and activation of canonical Wnt signaling during midfacial morphogenesis in mice. Dev Dyn 235:1448–1454

    Article  PubMed  CAS  Google Scholar 

  14. 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  PubMed  CAS  Google Scholar 

  15. Menezes R, Letra A, Kim AH, Kuchler EC, Day A, Tannure PN, Gomes da Motta L, Paiva KB, Granjeiro JM, Vieira AR (2010) Studies with Wnt genes and nonsyndromic cleft lip and palate. Birth Defects Res A Clin Mol Teratol 88:995–1000

    Article  PubMed  CAS  Google Scholar 

  16. Meng L, Bian Z, Torensma R, Von den Hoff JW (2009) Biological mechanisms in palatogenesis and cleft palate. J Dent Res 88:22–33

    Article  PubMed  CAS  Google Scholar 

  17. Mossey P (2007) Epidemiology underpinning research in the aetiology of orofacial clefts. Orthod Craniofac Res 10:114–120

    Article  PubMed  Google Scholar 

  18. Mossey PA, Little J, Munger RG, Dixon MJ, Shaw WC (2009) Cleft lip and palate. Lancet 374:1773–1785

    Article  PubMed  Google Scholar 

  19. Ngai CW, Martin WL, Tonks A, Wyldes MP, Kilby MD (2005) Are isolated facial cleft lip and palate associated with increased perinatal mortality? A cohort study from the West Midlands Region, 1995–1997. J Matern Fetal Neonatal Med 17:203–206

    Article  PubMed  Google Scholar 

  20. Song L, Li Y, Wang K, Wang YZ, Molotkov A, Gao L, Zhao T, Yamagami T, Wang Y, Gan Q et al (2009) Lrp6-mediated canonical Wnt signaling is required for lip formation and fusion. Development 136:3161–3171

    Article  PubMed  CAS  Google Scholar 

  21. Tyler MS, Pratt RM (1980) Effect of epidermal growth factor on secondary palatal epithelium in vitro: tissue isolation and recombination studies. J Embryol Exp Morphol 58:93–106

    PubMed  CAS  Google Scholar 

  22. Ueno S, Weidinger G, Osugi T, Kohn AD, Golob JL, Pabon L, Reinecke H, Moon RT, Murry CE (2007) Biphasic role for Wnt/beta-catenin signaling in cardiac specification in zebrafish and embryonic stem cells. Proc Natl Acad Sci U S A 104:9685–9690

    Article  PubMed  CAS  Google Scholar 

  23. Warner DR, Smith HS, Webb CL, Greene RM, Pisano MM (2009) Expression of Wnts in the developing murine secondary palate. Int J Dev Biol 53:1105–1112

    Article  PubMed  CAS  Google Scholar 

  24. Wehby GL, Cassell CH (2010) The impact of orofacial clefts on quality of life and healthcare use and costs. Oral Dis 16:3–10

    Article  PubMed  CAS  Google Scholar 

  25. Yao T, Yang L, Li PQ, Wu H, Xie HB, Shen X, Xie XD (2011) Association of Wnt3A gene variants with non-syndromic cleft lip with or without cleft palate in Chinese population. Arch Oral Biol 56:73–78

    Article  PubMed  CAS  Google Scholar 

  26. Zhu P, Chen G, You T, Yao J, Jiang Q, Lin X, Shen X, Qiao Y, Lin L (2010) High FFA-induced proliferation and apoptosis in human umbilical vein endothelial cell partly through Wnt/beta-catenin signal pathway. Mol Cell Biochem 338:123–131

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by a grant from the Science and Technology Project of Liaoning Province (grant no.: 2010225034).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Li Lu.

Electronic supplementary materials

Below is the link to the electronic supplementary material.

Supplemental Fig. 1

(JPEG 14 kb)

High resolution image (TIFF 728 kb)

Supplemental Table 1

(DOC 45 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Feng, C., Xu, Z., Li, Z. et al. Down-regulation of Wnt10a by RNA interference inhibits proliferation and promotes apoptosis in mouse embryonic palatal mesenchymal cells through Wnt/β-catenin signaling pathway. J Physiol Biochem 69, 855–863 (2013). https://doi.org/10.1007/s13105-013-0262-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13105-013-0262-7

Keywords

Navigation