Abstract
Previous studies indicate that Krüppel-like factor 5 (KLF5), also known as intestinal-enriched Krüppel-like factor (IKLF), is a positive regulator of cell proliferation and gives rise to a transformed phenotype when overexpressed. Here we demonstrate that levels of KLF5 transcript and protein are significantly elevated in oncogenic H-Ras-transformed NIH3T3 cells. These cells display an accelerated rate of proliferation in both serum-containing and serum-deprived media and form anchorage-independent colonies in soft agar assays. H-Ras-transformed cells also contain elevated mitogen-activated protein kinase (MAPK) activity. When treated with inhibitors of MEK (MAPK kinase), H-Ras-transformed cells lose their growth advantage and no longer form colonies. Significantly, levels of KLF5 transcript and protein are substantially reduced in H-Ras-transformed cells treated with MEK inhibitors. Moreover, inhibition of KLF5 expression in H-Ras-transformed cells with KLF5-specific small interfering RNA (siRNA) leads to a decreased rate of proliferation and a significant reduction in colony formation. H-Ras-transformed cells also contain elevated levels of Egr1 that are diminished by MEK inhibitors. Inhibition of Egr1 by siRNA results in a reduced level of KLF5, indicating that Egr1 mediates the inductive action of MAPK on KLF5. Lastly, KLF5 activates expression of cyclin D1. These findings indicate that the increased expression of KLF5 in H-Ras-transformed cells is secondary to increased MAPK activity from H-Ras overexpression and that the elevated level of KLF5 is in part responsible for the proproliferative and transforming activities of oncogenic H-Ras.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Abbreviations
- DMEM:
-
Dulbecco's modified Eagle's medium
- ERK:
-
extracellular signal-regulated kinase
- FBS:
-
fetal bovine serum
- GAPDH:
-
glyceraldehydes-3-phosphate dehydrogenase
- GKLF:
-
gut-enriched Krüppel-like factor
- IKLF:
-
intestinal Krüppel-like factor
- KLF:
-
Krüppel-like factor
- MAPK:
-
mitogen-activated protein kinase
- MEK:
-
MAPK/ERK kinase
- PBS:
-
phosphate-buffered saline
- PI:
-
propidium iodide
- PMA:
-
phorbol-12-myristate-13-acetate
- RT–PCR:
-
reverse transcription–polymerase chain reaction
- siRNA:
-
small interfering RNA
References
Barbacid M . (1987). Annu. Rev. Biochem., 56, 779–827.
Black AR, Black JD and Azizkhan-Clifford J . (2001). J. Cell. Physiol., 188, 143–160.
Bos JL, Fearon ER, Hamilton SR, Verlaan-de Vries M, van Boom JH, van der Eb AJ and Vogelstein B . (1987). Nature, 327, 293–297.
Chen C, Bhalala HV, Qiao H and Dong JT . (2002). Oncogene, 21, 6567–6572.
Chen C, Bhalala HV, Vessella RL and Dong JT . (2003). Prostate, 55, 81–88.
Chen JY, Penco S, Ostrowski J, Balaguer P, Pons M, Starrett JE, Reczek P, Chambon P and Gronemeyer H . (1995). EMBO J., 14, 1187–1197.
Cleveland JL, Troppmair J, Packham G, Askew DS, Lloyd P, Gonzalez-Garcia M, Nunez G, Ihle JN and Rapp UR . (1994). Oncogene, 9, 2217–2226.
Conkright MD, Wani MA, Anderson KP and Lingrel JB . (1999). Nucleic Acids Res., 27, 1263–1270.
Crossley M, Whitelaw E, Perkins A, Williams G, Fujiwara Y and Orkin SH . (1996). Mol. Cell. Biol., 16, 1695–1705.
Dang DT, Pevsner J and Yang VW . (2000). Int. J. Biochem. Cell. Biol., 32, 1103–1121.
Dang DT, Zhao W, Mahatan CS, Geiman DE and Yang VW . (2002). Nucleic Acids Res., 30, 2736–2741.
Daum G, Eisenmann-Tappe I, Fries HW, Troppmair J and Rapp UR . (1994). Trends Biochem. Sci., 19, 474–480.
de Vries-Smits AM, Burgering BM, Leevers SJ, Marshall CJ and Bos JL . (1992). Nature, 357, 602–604.
Dekker LV and Parker PJ . (1994). Trends Biochem. Sci., 19, 73–77.
DeSilva DR, Jones EA, Favata MF, Jaffee BD, Magolda RL, Trzaskos JM and Scherle PA . (1998). J. Immunol., 160, 4175–4181.
Dobrowolski S, Harter M and Stacey DW . (1994). Mol. Cell. Biol., 14, 5441–5449.
Downward J . (2003). Nat. Rev. Cancer, 3, 11–22.
Dudley DT, Pang L, Decker SJ, Bridges AJ and Saltiel AR . (1995). Proc. Natl. Acad. Sci. USA, 92, 7686–7689.
El-Shemerly MY, Besser D, Nagasawa M and Nagamine Y . (1997). J. Biol. Chem., 272, 30599–30602.
Favata MF, Horiuchi KY, Manos EJ, Daulerio AJ, Stradley DA, Feeser WS, Van Dyk DE, Pitts WJ, Earl RA, Hobbs F, Copeland RA, Magolda RL, Scherle PA and Trzaskos JM . (1998). J. Biol. Chem., 273, 18623–18632.
Filmus J, Robles AI, Shi W, Wong MJ, Colombo LL and Conti CJ . (1994). Oncogene, 9, 3627–3633.
Gallego C, Gupta SK, Heasley LE, Qian NX and Johnson GL . (1992). Proc. Natl. Acad. Sci. USA, 89, 7355–7359.
Garrett-Sinha LA, Eberspaecher H, Seldin MF and de Crombrugghe B . (1996). J. Biol. Chem., 271, 31384–31390.
Gebelein B, Fernandez-Zapico M, Imoto M and Urrutia R . (1998). J. Clin. Invest., 102, 1911–1919.
Hoshino Y, Kurabayashi M, Kanda T, Hasegawa A, Sakamoto H, Okamoto E, Kowase K, Watanabe N, Manabe I, Suzuki T, Nakano A, Takase S, Wilcox JN and Nagai R . (2000). Circulation, 102, 2528–2534.
Howe LR, Leevers SJ, Gomez N, Nakielny S, Cohen P and Marshall CJ . (1992). Cell, 71, 335–342.
Kaczynski J, Cook T and Urrutia R . (2003). Genome Biol., 4, 206.
Kawai-Kowase K, Kurabayashi M, Hoshino Y, Ohyama Y and Nagai R . (1999). Circ. Res., 85, 787–795.
Kerkhoff E and Rapp UR . (1998). Oncogene, 17, 1457–1462.
Khosravi-Far R, Campbell S, Rossman KL and Der CJ . (1998). Adv. Cancer Res., 72, 57–107.
Kolch W, Heidecker G, Kochs G, Hummel R, Vahidi H, Mischak H, Finkenzeller G, Marme D and Rapp UR . (1993). Nature, 364, 249–252.
Kullmann M, Gopfert U, Siewe B and Hengst L . (2002). Genes Dev., 16, 3087–3099.
Kuo ML, Kang JJ and Yang NC . (1993). Cancer Lett., 74, 197–202.
Leevers SJ, Paterson HF and Marshall CJ . (1994). Nature, 369, 411–414.
Liu JJ, Chao JR, Jiang MC, Ng SY, Yen JJ and Yang-Yen HF . (1995). Mol. Cell. Biol., 15, 3654–3663.
Macdonald SG, Crews CM, Wu L, Driller J, Clark R, Erikson RL and McCormick F . (1993). Mol. Cell. Biol., 13, 6615–6620.
Malumbres M and Pellicer A . (1998). Front Biosci., 3, d887–d912.
Marais R, Light Y, Paterson HF and Marshall CJ . (1995). EMBO J., 14, 3136–3145.
Ming XF, Burgering BM, Wennstrom S, Claesson-Welsh L, Heldin CH, Bos JL, Kozma SC and Thomas G . (1994). Nature, 371, 426–429.
Muszynski KW, Ruscetti FW, Heidecker G, Rapp U, Troppmair J, Gooya JM and Keller JR . (1995). J. Exp. Med., 181, 2189–2199.
Nori M, L’Allemain G and Weber MJ . (1992). Mol. Cell. Biol., 12, 936–945.
Ogata T, Kurabayashi M, Hoshino Y, Sekiguchi K, Ishikawa S, Morishita Y and Nagai R . (2000). J. Thorac. Cardiovasc. Surg., 119, 983–989.
Ohnishi S, Laub F, Matsumoto N, Asaka M, Ramirez F, Yoshida T and Terada M . (2000). Dev. Dyn., 217, 421–429.
Philipsen S and Suske G . (1999). Nucleic Acids Res., 27, 2991–3000.
Rajendran RR, Nye AC, Frasor J, Balsara RD, Martini PG and Katzenellenbogen BS . (2003). J. Biol. Chem., 278, 4628–4638.
Robbins DJ, Cheng M, Zhen E, Vanderbilt CA, Feig LA and Cobb MH . (1992). Proc. Natl. Acad. Sci. USA, 89, 6924–6928.
Shi H, Zhang Z, Wang X, Liu S and Teng CT . (1999). Nucleic Acids Res., 27, 4807–4815.
Shie JL, Chen ZY, O’Brien MJ, Pestell RG, Lee ME and Tseng CC . (2000). Am. J. Physiol. Gastrointest. Liver Physiol., 279, G806–G814.
Shields JM, Christy RJ and Yang VW . (1996). J. Biol. Chem., 271, 20009–20017.
Shields JM, Pruitt K, McFall A, Shaub A and Der CJ . (2000). Trends Cell. Biol., 10, 147–154.
Simmen RC and Simmen FA . (2002). Front Biosci., 7, d1556–d1565.
Sun R, Chen X and Yang VW . (2001). J. Biol. Chem., 276, 6897–6900.
Thomas SM, DeMarco M, D’Arcangelo G, Halegoua S and Brugge JS . (1992). Cell, 68, 1031–1040.
Troppmair J, Bruder JT, Munoz H, Lloyd PA, Kyriakis J, Banerjee P, Avruch J and Rapp UR . (1994). J. Biol. Chem., 269, 7030–7035.
Turner J and Crossley M . (1999). Trends Biochem. Sci., 24, 236–240.
Vogelstein B, Fearon ER, Hamilton SR, Kern SE, Preisinger AC, Leppert M, Nakamura Y, White R, Smits AM and Bos JL . (1988). N. Engl. J. Med., 319, 525–532.
Wartmann M, Hofer P, Turowski P, Saltiel AR and Hynes NE . (1997). J. Biol. Chem., 272, 3915–3923.
Watanabe G, Albanese C, Lee RJ, Reutens A, Vairo G, Henglein B and Pestell RG . (1998). Mol. Cell. Biol., 18, 3212–3222.
Winston JT, Coats SR, Wang YZ and Pledger WJ . (1996). Oncogene, 12, 127–134.
Yang JJ, Kang JS and Krauss RS . (1998). Mol. Cell. Biol., 18, 2586–2595.
Yoon HS, Chen X and Yang VW . (2003). J. Biol. Chem., 278, 2101–2105.
Ziemer LT, Pennica D and Levine AJ . (2001). Mol. Cell. Biol., 21, 562–574.
Acknowledgements
We thank Drs Raul Urrutia, Jerry Lingrel and Richard Pestell for kindly providing the various plasmid constructs in the study. This work was in part supported by grants from the National Institutes of Health (DK52230, DK64399 and CA84197). VWY is a recipient of a Georgia Cancer Coalition Distinguished Cancer Clinician Scientist Award.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nandan, M., Yoon, H., Zhao, W. et al. Krüppel-like factor 5 mediates the transforming activity of oncogenic H-Ras. Oncogene 23, 3404–3413 (2004). https://doi.org/10.1038/sj.onc.1207397
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1207397
Keywords
This article is cited by
-
KLF5-mediated Eppk1 expression promotes cell proliferation in cervical cancer via the p38 signaling pathway
BMC Cancer (2021)
-
Network analysis of KLF5 targets showing the potential oncogenic role of SNHG12 in colorectal cancer
Cancer Cell International (2020)
-
KLF5 functions in proliferation, differentiation, and apoptosis of chicken satellite cells
3 Biotech (2019)
-
Metformin suppresses triple-negative breast cancer stem cells by targeting KLF5 for degradation
Cell Discovery (2017)
-
Interpreting transcriptional changes using causal graphs: new methods and their practical utility on public networks
BMC Bioinformatics (2016)