Skip to main content
SpringerLink
Log in

Effect of Kruppel-like factor 4 on Notch pathway in hepatic stellate cells

  • Published:
Journal of Huazhong University of Science and Technology [Medical Sciences] Aims and scope Submit manuscript

Abstract

The relationship between Kruppel-like factor 4 (KLF4) and the Notch pathway was determined to investigate the effect of KLF4 on the activation of hepatic stellate cells and underlying mechanisms. Fifty SPF BALB/c mice were randomly divided into two groups. A liver fibrosis model was established in 25 mice as the experimental group, and the remaining 25 mice served as controls. On the day 0, 7, 14, and 35, liver tissues were removed for immunofluorescent detection. The Notch pathway inhibitor DAPT was added to the primary original hepatic stellate cells, and KLF4 and Notch-associated factor expression was detected by qRT-PCR. Additionally, the hepatic stellate cell line LX-2 was used to establish control and experimental groups, and was cultured in vitro. LX-2 cells in the experimental groups were treated with DAPT and the Notch activator transforming growth factor-beta 1 separately, whereas those in the control group were given isotonic culture medium. After 48 h, KLF4 expression was examined by Western blotting. After transient transfection of LX-2 cells to increase KLF4, the expression of Notch factor was examined. Immunofluorescence analysis showed that, with the aggravation of liver fibrosis, the absorbance (A) values of KLF4 were decreased (day 0: 980.73±153.19; day 7: 1087.99±230.23; day 14: 390.95±93.56; day 35: 245.99±87.34). The expression of Notch pathway- related factors (Notch-1, Notch-2, and Jagged-1) in the hepatic stellate cell membrane was negatively correlated to KLF4 expression. With the increase of KLF4 expression, Notch-2 (0.73±0.13) and Jagged-1 (0.43±0.12) expression decreased, whereas Notch-1 level was not detectable. When the Notch pathway was inhibited, KLF4 levels generally increased (18.12±1.31). Our results indicate that KLF4 expression is negatively correlated to the Notch pathway in hepatic stellate cells, which may provide a reference for the treatment of hepatic fibrosis.

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. Wei D, Wang L, Kanai M, et al. KLF4 alpha up-regulation promotes cell cycle progression and reduces survival time of patients with pancreatic cancer. Gastroenterology, 2010,139(6):2135–2145

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Shie JL, Chen ZY, O’Brien MJ, et al. Role of gut-enriched Kruppel-like factor in colonic cell growth and differentiation. Am J Physiol Gastrointest Liver Physiol, 2000,279(4):G806–814

    CAS  PubMed  Google Scholar 

  3. Dang DT, Chen X, Feng J, et al. Overexpression of Kruppel-like factor 4 in the human colon cancer cell line RKO leads to reduced tumorigenecity. Oncogene, 2003,22(22):3424–3430

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Evans PM, Chen X, Zhang W, et al. KLF4 interacts with beta-catenin/TCF4 and blocks p300/CBP recruitment by beta-catenin. Mol Cell Biol, 2010,30(2):372–381

    Article  CAS  PubMed  Google Scholar 

  5. Wong CW, Hou PS, Tseng SF, et al. Kruppel-like transcription factor 4 contributes to maintenance of telomerase activity in stem cells. Stem Cells, 2010,28(9):1510–1517

    Article  CAS  PubMed  Google Scholar 

  6. Yoon HS, Yang VW. Requirement of Kruppel-like factor 4 in preventing entry into mitosis following DNA damage. J Biol Chem, 2004,279(6):5035–5041

    Article  CAS  PubMed  Google Scholar 

  7. Chen ZY, Wang X, Zhou Y, et al. Destabilization of Kruppel-like factor 4 protein in response to serum stimulation involves the ubiquitin-proteasome pathway. Cancer Res, 2005,65(22):10394–10400

    Article  CAS  PubMed  Google Scholar 

  8. Chen X, Whitney EM, Gao SY, et al. Transcriptional profiling of Kruppel-like factor 4 reveals a function in cell cycle regulation and epithelial differentiation. J Mol Biol, 2003,326(3):665–677

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Butko E, Pouget C, Traver D. Complex regulation of HSC emergence by the Notch signaling pathway. Dev Biol, 2016,409(1):129–138

    Article  CAS  PubMed  Google Scholar 

  10. Zheng H, Pritchard DM, Yang X, et al. KLF4 gene expression is inhibited by the notch signaling pathway that controls goblet cell differentiation in mouse gastrointestinal tract. Am J Physiol Gastrointest Liver Physiol, 2009,296(3):G490–498

    Article  CAS  PubMed  Google Scholar 

  11. Fre S, Huyghe M, Mourikis P, et al. Notch signals control the fate of immature progenitor cells in the intestine. Nature, 2005,435(7044):964–968

    Article  CAS  PubMed  Google Scholar 

  12. Brandao DF, Ramalho LN, Ramalho FS, et al. Liver cirrhosis and hepatic stellate cells. Acta Cir Bras, 2006,21(Suppl 1):54–57

    Article  PubMed  Google Scholar 

  13. Sun XF, Sun XH, Cheng SF, et al. Interaction of the transforming growth factor-ß and Notch signaling pathways in the regulation of granulosa cell proliferation. Reprod Fertil Dev, 2015, doi: 10.1071/RD14398 [Epub ahead of print]

    Google Scholar 

  14. McCanless JD, Cole JA, Slack SM, et al. Modeling nucleus pulposus regeneration in vitro: mesenchymal stem cells, alginate beads, hypoxia, bone morphogenetic protein-2, and synthetic peptide B2A. Spine (Phila Pa 1976). 2011,36(26):2275–2285

    Google Scholar 

  15. Blokzijl A, Dahlqvist C, Reissmann E, et al. Cross-talk between the Notch and TGF-beta signaling pathways mediated by interaction of the Notch intracellular domain with Smad3. J Cell Biol, 2003,163(4):723–728

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Varela I, Karagiannidou A, Oikonomakis V, et al. Generation of human beta-thalassemia induced pluripotent cell lines by reprogramming of bone marrow-derived mesenchymal stromal cells using modified mRNA. Cell Reprogram, 2014,16(6):447–455

    Article  CAS  PubMed  Google Scholar 

  17. Chitilian JM, Thillainadesan G, Manias JL, et al. Critical components of the pluripotency network are targets for the p300/CBP interacting protein (p/CIP) in embryonic stem cells. Stem Cells, 2014,32(1):204–215

    Article  CAS  PubMed  Google Scholar 

  18. Zhuang SJ, Li X, Li KQ, et al. Mechanism of JAK/STAT signal pathway and Notch signaling crosstalking in proliferation and differentiation of neural stem cells. J Int Pharm Res, 2013,40(6):743–746

    CAS  Google Scholar 

  19. Qin S, Zou Y, Zhang CL. Cross-talk between KLF4 and STAT3 regulates axon regeneration. Nat Commun, 2013,4:2633

    PubMed  PubMed Central  Google Scholar 

  20. Roshan S, Liu YY, Banafa A, et al. Fucoidan induces apoptosis of HepG2 cells by down-regulating p-Stat3. J Huazhong Univ Sci Technol [Med Sci], 2014,34(3):330–336

    Article  CAS  Google Scholar 

  21. Ema M, Mori D, Niwa H, et al. Kruppel-like factor 5 is essential for blastocyst development and the normal self-renewal of mouse ESCs. Cell Stem Cell, 2008,3(5):555–567

    Article  CAS  PubMed  Google Scholar 

  22. Ghaleb AM, Aggarwal G, Bialkowska AB, et al. Notch inhibits expression of the Kruppel-like factor 4 tumor suppressor in the intestinal epithelium. Mol Cancer Res, 2008,6(12):1920–1927

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Emergency Surgery, Union Hospital, Tongji Medical College, Huazhong Univerfsity of Science and Technology, Wuhan, 430022, China

    Yin-kai Xue  (薛寅凯), Jun Tan  (谭 俊), Dong-wei Dou  (窦东伟), Ding Chen  (陈 丁), Lu-jia Chen  (陈路佳), Huan-ping Ren  (任换平), Li-bo Chen  (陈立波), Xin-gao Xiong  (熊新高) & Hai Zheng  (郑 海)

Authors
  1. Yin-kai Xue  (薛寅凯)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun Tan  (谭 俊)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dong-wei Dou  (窦东伟)
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ding Chen  (陈 丁)
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lu-jia Chen  (陈路佳)
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Huan-ping Ren  (任换平)
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Li-bo Chen  (陈立波)
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xin-gao Xiong  (熊新高)
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hai Zheng  (郑 海)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Xin-gao Xiong  (熊新高) or Hai Zheng  (郑 海).

Additional information

This project was supported by the National Natural Science Foundation of China (No. 81071541).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xue, Yk., Tan, J., Dou, Dw. et al. Effect of Kruppel-like factor 4 on Notch pathway in hepatic stellate cells. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 36, 811–816 (2016). https://doi.org/10.1007/s11596-016-1667-7

Download citation

  • Received:

  • Revised:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11596-016-1667-7

Keywords

Search

Navigation