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Journal of Cancer Research and Clinical Oncology

, Volume 139, Issue 6, pp 1033–1042 | Cite as

KLF8 involves in TGF-beta-induced EMT and promotes invasion and migration in gastric cancer cells

  • Hui Zhang
  • Lili Liu
  • Yafang Wang
  • Guohong Zhao
  • Rougang Xie
  • Changhao Liu
  • Xiao Xiao
  • Kaichun Wu
  • Yongzhan Nie
  • Hongbo ZhangEmail author
  • Daiming FanEmail author
Original Paper

Abstract

Purpose

Krüppel-like factor 8 (KLF8), a downstream transcription factor of transforming growth factor-β1 (TGF-β1), has a role in tumorigenesis, tumor progress and epithelial-to-mesenchymal transition (EMT) induction. Recent studies mainly focused on its role in breast cancer and hepatocellular carcinoma; however, little is studied in gastric cancer. Here, we aim to explore whether KLF8 is involved in TGF-β1-induced EMT in gastric cancer cells.

Methods

Western blot and real-time PCR assays were used to detect the expression of KLF8, E-cadherin and vimentin in gastric cancer cell line SGC7901 treated with or without TGF-β1. The lentivirus-mediated RNA interference technique was used to knock down the expression of KLF8 in gastric cancer cell line SGC7901. In vitro, the ability of cell migration and invasion were measured by transwell and wound healing assays; the cell motility was detected by high content screening assay.

Results

TGF-β1 could induce EMT via down-regulating E-cadherin and up-regulating vimentin expression in gastric cancer cells. Further study found that TGF-β1 could induce KLF8 expression at the protein and mRNA levels in gastric cancer cells (P < 0.05). Western blot and real-time PCR assays found that small interference RNA (siRNA)-mediated KLF8 silence blocked TGF-β1-induced EMT-like transformation and subsequently reversed the loss of E-cadherin and gain of vimentin. In vitro, inhibition of KLF8 decreased TGF-β1-prompted cell migration, invasion and motility.

Conclusions

KLF8, a transcription factor, is involved in TGF-β1-induced EMT in gastric cancer cells and may be a novel therapeutic target for the treatment of gastric cancer.

Keywords

TGF-β1 KLF8 EMT siRNA Gastric cancer 

Abbreviations

KLF8

Krüppel-like factor 8

TGF-β1

Transforming growth factor-β

EMT

Epithelial–mesenchymal transition

siRNA

Small interference RNA

ECM

Extra cellular matrix

cDNA

Complementary DNA

mRNA

Messenger RNA

GFP

Green fluorescent protein

NC

Negative control

Notes

Acknowledgments

Grant sponsor: National Nature Science Foundation of China (Number: 81030044).

Conflict of interest

We have no conflict of interest to be disclosed.

References

  1. Chen G, Yang W, Jin W, Wang Y, Tao C, Yu Z (2012) Lentivirus-mediated gene silencing of KLF8 reduced the proliferation and invasion of gastric cancer cells. Mol Biol Rep 39(10):9809–9815Google Scholar
  2. Choi YH, Choi KC, Park YE (1997) Relationship of transforming growth factor beta 1 to angiogenesis in gastric carcinoma. J Korean Med Sci 12(5):427–432PubMedGoogle Scholar
  3. Dang DT, Pevsner J, Yang VW (2000) The biology of the mammalian Kruppel-like family of transcription factors. Int J Biochem Cell Biol 32(11–12):1103–1121PubMedCrossRefGoogle Scholar
  4. Eaton SA, Funnell AP, Sue N, Nicholas H, Pearson RC, Crossley M (2008) A network of Kruppel-like Factors (Klfs). Klf8 is repressed by Klf3 and activated by Klf1 in vivo. J Biol Chem 283(40):26937–26947PubMedCrossRefGoogle Scholar
  5. Fu H, Hu Z, Wen J, Wang K, Liu Y (2009) TGF-beta promotes invasion and metastasis of gastric cancer cells by increasing fascin1 expression via ERK and JNK signal pathways. Acta Biochim Biophys Sin 41(8):648–656PubMedCrossRefGoogle Scholar
  6. Fu WJ, Li JC, Wu XY, Yang ZB, Mo ZN, Huang JW, Xia GW, Ding Q, Liu KD, Zhu HG (2010) Small interference RNA targeting Kruppel-like factor 8 inhibits the renal carcinoma 786–0 cells growth in vitro and in vivo. J Cancer Res Clin Oncol 136(8):1255–1265PubMedCrossRefGoogle Scholar
  7. Grandclement C, Pallandre JR, Valmary DS, Viel E, Bouard A, Balland J, Remy-Martin JP, Simon B, Rouleau A, Boireau W, Klagsbrun M, Ferrand C, Borg C (2011) Neuropilin-2 expression promotes TGF-beta1-mediated epithelial to mesenchymal transition in colorectal cancer cells. PLoS ONE 6(7):e20444PubMedCrossRefGoogle Scholar
  8. Holian J, Qi W, Kelly DJ, Zhang Y, Mreich E, Pollock CA, Chen XM (2008) Role of Kruppel-like factor 6 in transforming growth factor-beta1-induced epithelial-mesenchymal transition of proximal tubule cells. Am J Physiol Renal Physiol 295(5):F1388–F1396PubMedCrossRefGoogle Scholar
  9. Li X, Zhang YY, Wang Q, Fu SB (2005) Association between endogenous gene expression and growth regulation induced by TGF-beta1 in human gastric cancer cells. World J Gastroenterol 11(1):61–68PubMedGoogle Scholar
  10. Li JC, Yang XR, Sun HX, Xu Y, Zhou J, Qiu SJ, Ke AW, Cui YH, Wang ZJ, Wang WM, Liu KD, Fan J (2010) Up-regulation of Kruppel-like factor 8 promotes tumor invasion and indicates poor prognosis for hepatocellular carcinoma. Gastroenterology 139(6):2146–2157PubMedCrossRefGoogle Scholar
  11. Liu L, Liu N, Xu M, Liu Y, Min J, Pang H, Zhang N, Zhang H, Zhang H (2012) Lentivirus-delivered Kruppel-like factor 8 small interfering RNA inhibits gastric cancer cell growth in vitro and in vivo. Tumour Biol 33(1):53–61PubMedCrossRefGoogle Scholar
  12. Lv ZD, Na D, Liu FN, Du ZM, Sun Z, Li Z, Ma XY, Wang ZN, Xu HM (2010) Induction of gastric cancer cell adhesion through transforming growth factor-beta1-mediated peritoneal fibrosis. J Exp Clin Cancer Res 29:139PubMedCrossRefGoogle Scholar
  13. Mehta TS, Monzur F, Zhao J (2010) Determination of nuclear localization signal sequences for Kruppel-like factor 8. Methods Mol Biol 647:171–186PubMedCrossRefGoogle Scholar
  14. Meng X, Ezzati P, Wilkins JA (2011) Requirement of podocalyxin in TGF-beta induced epithelial mesenchymal transition. PLoS ONE 6(4):e18715PubMedCrossRefGoogle Scholar
  15. Miyazono K (2009) Transforming growth factor-beta signaling in epithelial-mesenchymal transition and progression of cancer. Proc Jpn Acad Ser B Phys Biol Sci 85(8):314–323PubMedCrossRefGoogle Scholar
  16. Schnell O, Romagna A, Jaehnert I, Albrecht V, Eigenbrod S, Juerchott K, Kretzschmar H, Tonn JC, Schichor C (2012) Kruppel-like factor 8 (KLF8) is expressed in gliomas of different WHO grades and is essential for tumor cell proliferation. PLoS ONE 7(1):e30429PubMedCrossRefGoogle Scholar
  17. Szkaradkiewicz A, Karpinski TM, Drews M, Borejsza-Wysocki M, Majewski P, Andrzejewska E (2010) Natural killer cell cytotoxicity and immunosuppressive cytokines (IL-10, TGF-beta1) in patients with gastric cancer. J Biomed Biotechnol 2010:901564PubMedCrossRefGoogle Scholar
  18. Thiery JP, Acloque H, Huang RY, Nieto MA (2009) Epithelial-mesenchymal transitions in development and disease. Cell 139(5):871–890PubMedCrossRefGoogle Scholar
  19. van Vliet J, Turner J, Crossley M (2000) Human Kruppel-like factor 8: a CACCC-box binding protein that associates with CtBP and represses transcription. Nucleic Acids Res 28(9):1955–1962PubMedCrossRefGoogle Scholar
  20. Wang X, Zhao J (2007) KLF8 transcription factor participates in oncogenic transformation. Oncogene 26(3):456–461PubMedCrossRefGoogle Scholar
  21. Wang X, Zheng M, Liu G, Xia W, McKeown-Longo PJ, Hung MC, Zhao J (2007) Kruppel-like factor 8 induces epithelial to mesenchymal transition and epithelial cell invasion. Cancer Res 67(15):7184–7193PubMedCrossRefGoogle Scholar
  22. Wang X, Urvalek AM, Liu J, Zhao J (2008) Activation of KLF8 transcription by focal adhesion kinase in human ovarian epithelial and cancer cells. J Biol Chem 283(20):13934–13942PubMedCrossRefGoogle Scholar
  23. Wei H, Wang X, Gan B, Urvalek AM, Melkoumian ZK, Guan JL, Zhao J (2006) Sumoylation delimits KLF8 transcriptional activity associated with the cell cycle regulation. J Biol Chem 281(24):16664–16671PubMedCrossRefGoogle Scholar
  24. Wu K, Liu BH, Zhao DY, Zhao Y (2001) Effect of vitamin E succinate on expression of TGF-beta1, c-Jun and JNK1 in human gastric cancer SGC-7901 cells. World J Gastroenterol 7(1):83–87PubMedGoogle Scholar
  25. Xiangming C, Natsugoe S, Takao S, Hokita S, Ishigami S, Tanabe G, Baba M, Kuroshima K, Aikou T (2001) Preserved Smad4 expression in the transforming growth factor beta signaling pathway is a favorable prognostic factor in patients with advanced gastric cancer. Clin Cancer Res 7(2):277–282PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Hui Zhang
    • 1
  • Lili Liu
    • 2
  • Yafang Wang
    • 1
  • Guohong Zhao
    • 1
  • Rougang Xie
    • 3
  • Changhao Liu
    • 1
  • Xiao Xiao
    • 1
  • Kaichun Wu
    • 1
  • Yongzhan Nie
    • 1
  • Hongbo Zhang
    • 1
    Email author
  • Daiming Fan
    • 1
    Email author
  1. 1.State Key Laboratory of Cancer Biology, Institute of Digestive Diseases, Xijing HospitalFourth Military Medical UniversityXi’anChina
  2. 2.Department of Oncology, Tangdu HospitalFourth Military Medical UniversityXi’anChina
  3. 3.Institute of NeuroscienceThe Fourth Military Medical UniversityXi’anChina

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