Skip to main content
SpringerLink
Log in

Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout

  • Research Article
  • Published:
Genes & Genomics Aims and scope Submit manuscript

Abstract

Background

Fam20c is intimately related to tissue development and diseases. At present, it has been reported that Fam20c regulates the mineralization of osteoblasts, but there are few reports on other effects.

Objective

To study the effect of Fam20c on osteoblasts by knocking out the Fam20c gene.

Methods

Fam20c knockout osteoblasts were constructed by transfecting mouse osteoblasts with lentivirus. The proliferation, migration and mineralization of Fam20c knockout cells were detected by CCK-8, scratch test and alizarin red staining assays. The subcellular structure was observed by transmission electron microscopy. RT–PCR was used to detect the differential expression of mesenchymal-to-epithelial transition (MET)-related marker genes and core transcription factors. The differential expression of MET-related proteins was detected by immunofluorescence or Western blot. Transcriptome analysis of Fam20c knockout osteoblasts was performed, and real-time PCR was used to verify transcriptome analysis related to MET.

Results

The proliferation ability of osteoblasts was not significantly changed after Fam20c deletion, but the migration ability and mineralization ability were significantly weakened. There were tight junctions between Fam20c knockout cells. The expression of mesenchymal cell marker genes and core transcription factors was significantly decreased, and the expression of epithelial cell marker genes was significantly increased. The expression of mesenchymal cell marker proteins was significantly decreased, and the expression of epithelial cell marker proteins was significantly increased. Multiple signalling molecules and pathways involved in MET have changed.

Conclusions

Knockdown of Fam20c resulted in MET. Fam20c affects the transcription of key factors in osteoblast MET.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Availability of data and material

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

References

  • Andre P, Song H, Kim W, Kispert A, Yang Y (2015) Wnt5a and Wnt11 regulate mammalian anterior-posterior axis elongation. Development 142:1516–1527

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cai X, Nomura-Kitabayashi A, Cai W, Yan J, Christoffels VM, Cai CL (2011) Myocardial Tbx20 regulates early atrioventricular canal formation and endocardial epithelial-mesenchymal transition via Bmp2. Dev Biol 360:381–390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen T, You Y, Jiang H, Wang ZZ (2017) Epithelial-mesenchymal transition (EMT): a biological process in the development, stem cell differentiation, and tumorigenesis. J Cell Physiol 232:3261–3272

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cozza G, Moro E, Black M, Marin O, Salvi M, Venerando A, Tagliabracci VS, Pinna LA (2018) The Golgi ‘casein kinase’ Fam20C is a genuine ‘phosvitin kinase’ and phosphorylates polyserine stretches devoid of the canonical consensus. FEBS J 285:4674–4683

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • El-Dessouky SH, Abdel-Hamid MS, Abdel-Ghafar SF, Aboulghar MM, Gaafar HM, Fouad M, Ahmed AH, Abdel-Salam G (2020) Raine syndrome: prenatal diagnosis based on recognizable fetal facial features and characteristic intracranial calcification. Prenat Diagn 40:1578–1597

    Article  CAS  PubMed  Google Scholar 

  • Goettsch C, Hutcheson JD, Aikawa M, Iwata H, Pham T, Nykjaer A, Kjolby M, Rogers M, Michel T, Shibasaki M et al (2016) Sortilin mediates vascular calcification via its recruitment into extracellular vesicles. J Clin Invest 126:1323–1336

    Article  PubMed  PubMed Central  Google Scholar 

  • Hamidi S, Nakaya Y, Nagai H, Alev C, Kasukawa T, Chhabra S, Lee R, Niwa H, Warmflash A, Shibata T et al (2020) Mesenchymal-epithelial transition regulates initiation of pluripotency exit before gastrulation. Development 147:dev184960

    Article  CAS  PubMed  Google Scholar 

  • Hirose K, Ishimoto T, Usami Y, Sato S, Oya K, Nakano T, Komori T, Toyosawa S (2020) Overexpression of Fam20C in osteoblast in vivo leads to increased cortical bone formation and osteoclastic bone resorption. Bone 138:115414

    Article  CAS  PubMed  Google Scholar 

  • Huang JQ, Wei FK, Xu XL, Ye SX, Song JW, Ding PK, Zhu J, Li HF, Luo XP, Gong H et al (2019) SOX9 drives the epithelial-mesenchymal transition in non-small-cell lung cancer through the Wnt/β-catenin pathway. J Transl Med 17:143

    Article  PubMed  PubMed Central  Google Scholar 

  • Lamouille S, Xu J, Derynck R (2014) Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol 15:178–196

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee JM, Dedhar S, Kalluri R, Thompson EW (2006) The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol 172:973–981

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ligorio M, Sil S, Malagon-Lopez J, Nieman LT, Misale S, Di Pilato M, Ebright RY, Karabacak MN, Kulkarni AS, Liu A et al (2019) Stromal microenvironment shapes the intratumoral architecture of pancreatic cancer. Cell 178:160-175.e27

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu C, Wang X, Zhang H, Xie X, Liu P, Liu Y, Jani PH, Lu Y, Chen S, Qin C (2015) Immortalized mouse floxed Fam20c dental papillar mesenchymal and osteoblast cell lines retain their primary characteristics. J Cell Physiol 230:2581–2587

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu C, Zhang H, Jani P, Wang X, Lu Y, Li N, Xiao J, Qin C (2018) FAM20C regulates osteoblast behaviors and intracellular signaling pathways in a cell-autonomous manner. J Cell Physiol 233:3476–3486

    Article  CAS  PubMed  Google Scholar 

  • Ma L, Lu MF, Schwartz RJ, Martin JF (2005) Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning. Development 132:5601–5611

    Article  CAS  PubMed  Google Scholar 

  • Ma Z, Liu D, Li W, Di S, Zhang Z, Zhang J, Xu L, Guo K, Zhu Y, Han J et al (2019) STYK1 promotes tumor growth and metastasis by reducing SPINT2/HAI-2 expression in non-small cell lung cancer. Cell Death Dis 10:435

    Article  PubMed  PubMed Central  Google Scholar 

  • Mamedova E, Dimitrova D, Przhiyalkovskaya E, Buryakina S, Vasilyev E, Tiulpakov A, Belaya Z (2019) Non-lethal raine syndrome in a middle-aged woman caused by a novel FAM20C mutation. Calcif Tissue Int 105:567–572

    Article  CAS  PubMed  Google Scholar 

  • Miao N, Zhan Y, Xu Y, Yuan H, Qin C, Lin F, Xie X, Mu S, Yuan M, Mu H et al (2019) Loss of Fam20c causes defects in the acinar and duct structure of salivary glands in mice. Int J Mol Med 43:2103–2117

    CAS  PubMed  PubMed Central  Google Scholar 

  • Nakaya Y, Kuroda S, Katagiri YT, Kaibuchi K, Takahashi Y (2004) Mesenchymal-epithelial transition during somitic segmentation is regulated by differential roles of Cdc42 and Rac1. Dev Cell 7:425–438

    Article  CAS  PubMed  Google Scholar 

  • Nalbant D, Youn H, Nalbant SI, Sharma S, Cobos E, Beale EG, Du Y, Williams SC (2005) FAM20: an evolutionarily conserved family of secreted proteins expressed in hematopoietic cells. BMC Genomics 6:11

    Article  PubMed  PubMed Central  Google Scholar 

  • Okagawa H, Markwald RR, Sugi Y (2007) Functional BMP receptor in endocardial cells is required in atrioventricular cushion mesenchymal cell formation in chick. Dev Biol 306:179–192

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pastushenko I, Blanpain C (2019) EMT transition states during tumor progression and metastasis. Trends Cell Biol 29:212–226

    Article  CAS  PubMed  Google Scholar 

  • Pei D, Shu X, Gassama-Diagne A, Thiery JP (2019) Mesenchymal-epithelial transition in development and reprogramming. Nat Cell Biol 21:44–53

    Article  CAS  PubMed  Google Scholar 

  • Pollak AJ, Haghighi K, Kunduri S, Arvanitis DA, Bidwell PA, Liu GS, Singh VP, Gonzalez DJ, Sanoudou D, Wiley SE et al (2011) p53 coordinates cranial neural crest cell growth and epithelial-mesenchymal transition/delamination processes. Development 138:1827–1838

    Article  Google Scholar 

  • Rinon A, Molchadsky A, Nathan E, Yovel G, Rotter V, Sarig R, Tzahor E (2011) p53 coordinates cranial neural crest cell growth and epithelial-mesenchymal transition/delamination processes. Development 138:1827–1838

    Article  CAS  PubMed  Google Scholar 

  • Rios Garcia M, Steinbauer B, Srivastava K, Singhal M, Mattijssen F, Maida A, Christian S, Hess-Stumpp H, Augustin HG, Müller-Decker K et al (2017) Acetyl-CoA carboxylase 1-dependent protein acetylation controls breast cancer metastasis and recurrence. Cell Metab 26:842-855.e5

    Article  CAS  PubMed  Google Scholar 

  • Stemmler MP, Eccles RL, Brabletz S, Brabletz T (2019) Non-redundant functions of EMT transcription factors. Nat Cell Biol 21:102–112

    Article  CAS  PubMed  Google Scholar 

  • Tagliabracci VS, Engel JL, Wen J, Wiley SE, Worby CA, Kinch LN, Xiao J, Grishin NV, Dixon JE (2012) Secreted kinase phosphorylates extracellular proteins that regulate biomineralization. Science 336:1150–1153

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tagliabracci VS, Wiley SE, Guo X, Kinch LN, Durrant E, Wen J, Xiao J, Cui J, Nguyen KB, Engel JL et al (2015) A single kinase generates the majority of the secreted phosphoproteome. Cell 161:1619–1632

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Takahashi Y, Sato Y, Suetsugu R, Nakaya Y (2005) Mesenchymal-to-epithelial transition during somitic segmentation: a novel approach to studying the roles of Rho family GTPases in morphogenesis. Cells Tissues Organs 179:36–42

    Article  CAS  PubMed  Google Scholar 

  • Takeichi M, Nakagawa S, Aono S, Usui T, Uemura T (2000) Patterning of cell assemblies regulated by adhesion receptors of the cadherin superfamily. Philos Trans R Soc Lond B Biol Sci 355:885–890

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Takeyari S, Yamamoto T, Kinoshita Y, Fukumoto S, Glorieux FH, Michigami T, Hasegawa K, Kitaoka T, Kubota T, Imanishi Y et al (2014) Hypophosphatemic osteomalacia and bone sclerosis caused by a novel homozygous mutation of the FAM20C gene in an elderly man with a mild variant of Raine syndrome. Bone 67:56–62

    Article  CAS  PubMed  Google Scholar 

  • Thiery JP, Acloque H, Huang RY, Nieto MA (2009) Epithelial-mesenchymal transitions in development and disease. Cell 139:871–890

    Article  CAS  PubMed  Google Scholar 

  • Wiggan O, Hamel PA (2002) Pax3 regulates morphogenetic cell behavior in vitro coincident with activation of a PCP/non-canonical Wnt-signaling cascade. J Cell Sci 115:531–541

    Article  CAS  PubMed  Google Scholar 

  • Xie Y, Lu W, Liu S, Yang Q, Carver BS, Li E, Wang Y, Fazli L, Gleave M, Chen Z (2014) Crosstalk between nuclear MET and SOX9/β-catenin correlates with castration-resistant prostate cancer. Mol Endocrinol 28:1629–1639

    Article  PubMed  PubMed Central  Google Scholar 

  • Yang YJ, Luo S, Wang LS (2019) Effects of microRNA-378 on epithelial-mesenchymal transition, migration, invasion and prognosis in gastric carcinoma by targeting BMP2. Eur Rev Med Pharmacol Sci 23:5176–5186

    PubMed  Google Scholar 

  • Yook JI, Li XY, Ota I, Hu C, Kim HS, Kim NH, Cha SY, Ryu JK, Choi YJ, Kim J et al (2006) A Wnt-Axin2-GSK3beta cascade regulates Snail1 activity in breast cancer cells. Nat Cell Biol 8:1398–1406

    Article  CAS  PubMed  Google Scholar 

  • Zeisberg M, Neilson EG (2009) Biomarkers for epithelial-mesenchymal transitions. J Clin Invest 119:1429–1437

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang H, Zhu Q, Cui J, Wang Y, Chen MJ, Guo X, Tagliabracci VS, Dixon JE, Xiao J (2018a) Structure and evolution of the Fam20 kinases. Nat Commun 9:1218

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang J, Zhu Q, Wang X, Yu J, Chen X, Wang J, Wang X, Xiao J, Wang CC, Wang L (2018) Secretory kinase Fam20C tunes endoplasmic reticulum redox state via phosphorylation of Ero1α. EMBO J 37:e98699

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhou H, Li G, Huang S, Feng Y, Zhou A (2019) SOX9 promotes epithelial-mesenchymal transition via the Hippo-YAP signaling pathway in gastric carcinoma cells. Oncol Lett 18:599–608

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by National Natural Science Foundation of China (81870736, 81801040, 81500816, 81570951).

Author information

Author notes

  1. Zhen Zhang and Ya-Wei Geng have contributed equally to this work.

Authors and Affiliations

  1. Institute of Hard Tissue Development and Regeneration, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, Heilongjiang, People’s Republic of China

    Ya-Wei Geng, Zhen Zhang, Han Jin, Jun-Long Da, Kai Zhang, Jian-Qun Wang, Yu-Yao Guo, Bin Zhang & Ying Li

  2. Heilongjiang Provincial Key Laboratory of Hard Tissue Development and Regeneration, Harbin, 150001, Heilongjiang, People’s Republic of China

    Ya-Wei Geng, Zhen Zhang, Han Jin, Jun-Long Da, Kai Zhang, Jian-Qun Wang, Yu-Yao Guo, Bin Zhang & Ying Li

  3. Heilongjiang Academy of Medical Sciences, Harbin, 150001, Heilongjiang, People’s Republic of China

    Bin Zhang

Authors
  1. Ya-Wei Geng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhen Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Han Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jun-Long Da
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kai Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jian-Qun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yu-Yao Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Bin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ying Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Bin Zhang or Ying Li.

Ethics declarations

Conflict of interest

Author Ya-Wei Geng, author Zhen Zhang, author Han Jin, author Jun-Long Da, author Kai Zhang, author Jian-Qun Wang, author Yu-Yao Guo, author Bin Zhang, and author Ying Li declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Geng, YW., Zhang, Z., Jin, H. et al. Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout. Genes Genom 44, 155–164 (2022). https://doi.org/10.1007/s13258-021-01170-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13258-021-01170-4

Keywords

Search

Navigation