Skip to main content

Advertisement

SpringerLink
Log in

Induction of claudins in passaged hTERT-transfected human nasal epithelial cells with an extended life span

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

The epithelial barrier of the upper respiratory tract, such as that of the nasal mucosa, plays a crucial role in host defense. The epithelial barrier is regulated in large part by the apical-most intercellular junctions, referred to as tight junctions. However, the mechanisms regulating of tight junction barrier in human nasal epithelial cells remain unclear because the proliferation and storage of epithelial cells in primary cultures are limited. In the present study, we introduced the catalytic component of telomerase, the hTERT gene, into primary cultured human nasal epithelial cells and examined the properties of the transfectants, including their expression of tight junctions, compared with primary cultures. The ectopic expression of hTERT in the epithelial cells resulted in adequate growth potential and a longer lifespan of the cells. The properties of the passaged hTERT-transfected cells including tight junctions were similar to those of the cells in primary cultures. The barrier function in the transfectants after treatment with 10% FBS was significantly enhanced with increases of integral tight junction proteins claudin-1 and -4. When the transfectants were treated with TGF-β, which is assosciated with nasal polyposis and chronic rhinosinusitis, upregulation of only claudin-4 was observed, without a change of barrier function. In human nasal epithelial cells, the claudins may be important for barrier function and a novel target for a drug-delivery system. Our results indicate that hTERT-transfected human nasal epithelial cells with an extended lifespan can be used as an indispensable and stable model for studying the regulation of claudins in human nasal epithelium.

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
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Beitzinger M, Oswald C, Beinoraviciute-Kellner R, Stiewe T (2006) Regulation of telomerase activity by the p53 family member p73. Oncogene 25:813–826

    Article  PubMed  CAS  Google Scholar 

  • Bodnar AG, Ouellette M, Frolkis M, Holt EC, Chiu CP, Wright WE (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279:349–352

    Article  PubMed  CAS  Google Scholar 

  • Fujita K, Katahira J, Horiguchi Y, Sonoda N, Furuse M, Tsukita S (2000) Clostridium perfringens enterotoxin binds to the second extracellular loop of claudin-3, a tight junction integral membrane protein. FEBS Lett 476:258–261

    Article  PubMed  CAS  Google Scholar 

  • Gumbiner B (1993) Breaking through the tight junction barrier. J Cell Biol 123:1631–1633

    Article  PubMed  CAS  Google Scholar 

  • Herard AL, Zahm JM, Pierrot D, Hinnrasky J, Fuchey C, Puchelle E (1996) Epithelial barrier integrity during in vitro wound repair of the airway epithelium. Am J Respir Cell Mol Biol 15:624–632

    PubMed  CAS  Google Scholar 

  • Howe KL, Reardon C, Wang A, Nazli A, McKay DM (2005) Transforming growth factor-beta regulation of epithelial tight junction proteins enhances barrier function and blocksenterohemorrhagic Escherichia coli O157:H7-induced increased permeability. Am J Pathol 167:1587–1597

    PubMed  CAS  Google Scholar 

  • Katahira J, Sugiyama H, Inoue N, Horiguchi Y, Matsuda M, Sugimoto N (1997) Clostridium perfringens enterotoxin utilizes two structurally related membrane proteins as functional receptors in vivo. J Biol Chem 272:26652–26658

    Article  PubMed  CAS  Google Scholar 

  • Kawano Y, Kobune M, Yamaguchi M, Nakamura K, Ito Y, Sasaki K, Takahashi S, Nakamura T, Chiba H, Sato T, Matsunaga T, Azuma H, Ikebuchi K, Ikeda H, Kato J, Niitsu Y, Hamada H (2003) Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells. Blood 101:532–540

    Article  PubMed  CAS  Google Scholar 

  • Kondoh M, Takahashi A, Fujii M, Yagi K, Watanabe Y (2006) A novel strategy for a drug delivery system using a claudin modulator. Biol Pharm Bull 29:1783–1789

    Article  PubMed  CAS  Google Scholar 

  • Lin H, Yoo JW, Roh HJ, Lee MK, Chung SJ, Shim CK, Kim DD (2005) Transport of anti-allergic drugs across the passage cultured human nasal epithelial cell monolayer. Eur J Pharm Sci 26:203–210

    PubMed  CAS  Google Scholar 

  • McCarthy K, Francis S, McCormack J, Lai J, Rogers R, Skare I, Lynch R, Schneeberger E (2000) Inducible expression of claudin-1-myc but not occludin-VSV-G results in aberrant tight junction strand formation in MDCK cells. J Cell Sci 113:3387–3398

    PubMed  CAS  Google Scholar 

  • Medici D, Hay ED, Goodenough DA (2006) Cooperation between snail and LEF-1 transcription factors is essential for TGF-beta1-induced epithelial-mesenchymal transition. Mol Biol Cell 17:1871–1879

    Article  PubMed  CAS  Google Scholar 

  • Meyerson M (1998) Telomerase enzyme activation and human cell immortalization. Toxicol Lett 102–103:41–45

    Article  PubMed  Google Scholar 

  • Nisato RE, Harrison JA, Buser R, Orci L, Rinsch C, Montesano R, Dupraz P, Pepper MS (2004) Generation and characterization of telomerase-transfected human lymphatic endothelial cells with an extended life span. Am J Pathol 165:11–24

    PubMed  CAS  Google Scholar 

  • Pawankar R (2003) Nasal polyposis: an update: editorial review. Curr Opin Allergy Clin Immunol 3:1–6

    Article  PubMed  CAS  Google Scholar 

  • Piao CQ, Liu L, Zhao YL, Balajee AS, Suzuki M, Hei TK (2005) Immortalization of human small airway epithelial cells by ectopic expression of telomerase. Carcinogenesis 26:725–731

    Article  PubMed  CAS  Google Scholar 

  • Rufer N, Migliaccio M, Antonchuk J, Lansdorop PM (2001) Transfer of the human telomerase reverse transcriptase (TERT) gene into T lymphocytes results in extension of replicative potential. Blood 98:597–603

    Article  PubMed  CAS  Google Scholar 

  • Sawada N, Murata M, Kikuchi K, Tobioka H, Kojima T, Chiba H (2003) Tight junctions and human disease. Med Electron Microsc 36:147–156

    Article  PubMed  Google Scholar 

  • Schneeberger EE (2003) Claudins form ion-selective channels in the paracellular pathway. Focus on “Claudin extracellular domains determine paracellular charge selectively and resistance but not tight junction fibril architecture”. Am J Physiol Cell Physiol 284:C1331–C1333

    PubMed  CAS  Google Scholar 

  • Schneeberger EE, Lynch RD (1992) Structure, function, and regulation of cellular tight junctions. Am J Physiol 262:L647–L661

    PubMed  CAS  Google Scholar 

  • Schneeberger EE, Lynch RD (2004) The tight junction: a multifunctional complex. Am J Physiol 286:C1213–C1228

    Article  CAS  Google Scholar 

  • Sonoda N, Furuse M, Sasaki H, Yonemura S, Katahira J, Horiguchi Y, Tsukita S (1999) Clostridium perfringens enterotoxin fragment removes specific claudins from tight junction strands: evidence for direct involvement of claudins in tight junction barrier. J Cell Biol 147:195–204

    Article  PubMed  Google Scholar 

  • Takano K, Kojima T, Go M, Murata M, Ichimiya S, Himi T, Sawada N (2005) HLA-DR- and CD11c-positive dendritic cells penetrate beyond well-developed epithelial tight junctions in human nasal mucosa of allergic rhinitis. J Histochem Cytochem 53:611–619

    Article  PubMed  CAS  Google Scholar 

  • Tsukita S, Furuse M, Itoh M (2001) Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol 2:285–293

    Article  PubMed  CAS  Google Scholar 

  • Van Itallie CM, Anderson JM (2006) Claudins and epithelial paracellular transport. Annu Rev Physiol 68:403–429

    Article  PubMed  CAS  Google Scholar 

  • Van Itallie C, Rahner C, Anderson JM (2001) Regulated expression of claudin-4 decreases paracellular conductance through a selective decrease in sodium permeability. J Clin Invest 107:1319–1327

    Article  PubMed  Google Scholar 

  • Van Kempen MJP, Rijkers GT, Van Cauwenberge PB (2000) The immune response in adenoids and tonsils. Int Arch Allergy Immunol 122:8–19

    Article  PubMed  Google Scholar 

  • Vaziri H, Benchimol S (1998) Reconstitution of telomerase activity in normal human cells leads to elongation of telomeres and extended replicative life span. Curr Biol 8:279–282

    Article  PubMed  CAS  Google Scholar 

  • Yang J, Chang E, Cherry AM, Bangs CD, Herron GS (1999) Human endothelial cell life extension by telomerase expression. J Biol Chem 274:26141–26148

    Article  PubMed  CAS  Google Scholar 

  • Yoo JW, Kim YS, Lee SH, Lee MK, Roh HJ, Jhun BH, Lee CH, Kim DD (2003) Serially passaged human nasal epithelial cell monolayer for in vitro drug transport studies. Pharm Res 20:1690–1696

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Ms. E. Suzuki (Sapporo Medical University) for technical support.

Author information

Authors and Affiliations

  1. Department of Otolaryngology, Sapporo Medical University School of Medicine, Sapporo, Japan

    Makoto Kurose, Jun-ichi Koizumi, Ryuta Kamekura & Tetsuo Himi

  2. Department of Pathology, Sapporo Medical University School of Medicine, S1. W17, Sapporo, 060-8556, Japan

    Makoto Kurose, Takashi Kojima, Jun-ichi Koizumi, Ryuta Kamekura, Masaki Murata, Shingo Ichimiya, Makoto Osanai, Hideki Chiba & Norimasa Sawada

  3. Departments of Anatomy, Sapporo Medical University School of Medicine, Sapporo, Japan

    Takafumi Ninomiya

Authors
  1. Makoto Kurose
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Takashi Kojima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun-ichi Koizumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ryuta Kamekura
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takafumi Ninomiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Masaki Murata
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shingo Ichimiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Makoto Osanai
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hideki Chiba
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Tetsuo Himi
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Norimasa Sawada
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Takashi Kojima.

Additional information

This work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports Science, and Technology of Japan, the Ministry of Health, Labor, and Welfare of Japan, Japan Science and Technology Agency, the Akiyama Foundation, and the Long-Range Research Initiative Project of the Japan Chemical Industry Association.

Electronic supplementary material

Below is the link to the electronic supplementary material.

441_2007_453_MOESM1_ESM.pdf

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kurose, M., Kojima, T., Koizumi, Ji. et al. Induction of claudins in passaged hTERT-transfected human nasal epithelial cells with an extended life span. Cell Tissue Res 330, 63–74 (2007). https://doi.org/10.1007/s00441-007-0453-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-007-0453-z

Keywords

Search

Navigation