Abstract
KLF4 (GKLF/EZF) encodes a transcription factor that is associated with both tumour suppression and oncogenesis. We describe the identification of KLF4 in a functional genomic screen for genes that bypass RASV12-induced senescence. However, in untransformed cells, KLF4 acts as a potent inhibitor of proliferation. KLF4-induced arrest is bypassed by oncogenic RASV12 or by the RAS target cyclin-D1. Remarkably, inactivation of the cyclin-D1 target and the cell-cycle inhibitor p21CIP1 not only neutralizes the cytostatic action of KLF4, but also collaborates with KLF4 in oncogenic transformation. Conversely, KLF4 suppresses the expression of p53 by directly acting on its promoter, thereby allowing for RASV12-mediated transformation and causing resistance to DNA-damage-induced apoptosis. Consistently, KLF4 depletion from breast cancer cells restores p53 levels and causes p53-dependent apoptosis. These results unmask KLF4 as a regulator of p53 that oncogenically transforms cells as a function of p21CIP1 status. Furthermore, they provide a mechanistic explanation for the context-dependent oncogenic or tumour-suppressor functions of KLF4.
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References
Shields, J. M., Christy, R. J. & Yang, V. W. Identification and characterization of a gene encoding a gut-enriched Kruppel-like factor expressed during growth arrest. J. Biol. Chem. 271, 20009–20017 (1996).
Garrett-Sinha, L. A., Eberspaecher, H., Seldin, M. F. & de Crombrugghe, B. A gene for a novel zinc-finger protein expressed in differentiated epithelial cells and transiently in certain mesenchymal cells. J. Biol. Chem. 271, 31384–31390 (1996).
Chen, X., Whitney, E. M., Gao, S. Y. & Yang, V. W. Transcriptional profiling of Kruppel-like factor 4 reveals a function in cell cycle regulation and epithelial differentiation. J. Mol. Biol. 326, 665–677 (2003).
Chen, X. et al. Kruppel-like factor 4 (gut-enriched Kruppel-like factor) inhibits cell proliferation by blocking G1/S progression of the cell cycle. J. Biol. Chem. 276, 30423–30428 (2001).
Geiman, D. E., Ton-That, H., Johnson, J. M. & Yang, V. W. Transactivation and growth suppression by the gut-enriched Kruppel-like factor (Kruppel-like factor 4) are dependent on acidic amino acid residues and protein-protein interaction. Nucleic Acids Res. 28, 1106–1113 (2000).
Zhang, W. et al. The gut-enriched Kruppel-like factor (Kruppel-like factor 4) mediates the transactivating effect of p53 on the p21WAF1/Cip1 promoter. J. Biol. Chem. 275, 18391–18398 (2000).
Jaubert, J., Cheng, J. & Segre, J. A. Ectopic expression of kruppel like factor 4 (Klf4) accelerates formation of the epidermal permeability barrier. Development 130, 2767–2777 (2003).
Katz, J. P. et al. The zinc-finger transcription factor Klf4 is required for terminal differentiation of goblet cells in the colon. Development 129, 2619–2628 (2002).
Segre, J. A., Bauer, C. & Fuchs, E. Klf4 is a transcription factor required for establishing the barrier function of the skin. Nature Genet. 22, 356–360 (1999).
Black, A. R., Black, J. D. & Azizkhan-Clifford, J. Sp1 and kruppel-like factor family of transcription factors in cell growth regulation and cancer. J. Cell Physiol. 188, 143–160 (2001).
Ohnishi, S. et al. Downregulation and growth inhibitory effect of epithelial-type Kruppel-like transcription factor KLF4, but not KLF5, in bladder cancer. Biochem. Biophys. Res. Commun. 308, 251–256 (2003).
Dang, D. T., Mahatan, C. S., Dang, L. H., Agboola, I. A. & Yang, V. W. Expression of the gut-enriched Kruppel-like factor (Kruppel-like factor 4) gene in the human colon cancer cell line RKO is dependent on CDX2. Oncogene 20, 4884–4890 (2001).
Dang, D. T. et al. Decreased expression of the gut-enriched Kruppel-like factor gene in intestinal adenomas of multiple intestinal neoplasia mice and in colonic adenomas of familial adenomatous polyposis patients. FEBS Lett. 476, 203–207 (2000).
Ton-That, H., Kaestner, K. H., Shields, J. M., Mahatanankoon, C. S. & Yang, V. W. Expression of the gut-enriched Kruppel-like factor gene during development and intestinal tumorigenesis. FEBS Lett. 419, 239–243 (1997).
Shie, J. L. et al. Role of gut-enriched Kruppel-like factor in colonic cell growth and differentiation. Am. J. Physiol. Gastrointest. Liver Physiol. 279, G806–G814 (2000).
Zhao, W. et al. Identification of Kruppel-like factor 4 as a potential tumor suppressor gene in colorectal cancer. Oncogene 23, 395–402 (2004).
Wei, D. et al. Drastic down-regulation of Kruppel-like factor 4 expression is critical in human gastric cancer development and progression. Cancer Res. 65, 2746–2754 (2005).
Katz, J. P. et al. Loss of Klf4 in mice causes altered proliferation and differentiation and precancerous changes in the adult stomach. Gastroenterology 128, 935–945 (2005).
Luo, A. et al. Discovery of Ca(2+)-relevant and differentiation-associated genes downregulated in esophageal squamous cell carcinoma using cDNA microarray. Oncogene 1, 1 (2003).
Dang, D. T. et al. Overexpression of Kruppel-like factor 4 in the human colon cancer cell line RKO leads to reduced tumorigenecity. Oncogene 22, 3424–3430 (2003).
Foster, K. W. et al. Increase of GKLF messenger RNA and protein expression during progression of breast cancer. Cancer Res. 60, 6488–6495 (2000).
Foster, K. W. et al. Oncogene expression cloning by retroviral transduction of adenovirus E1A-immortalized rat kidney RK3E cells: transformation of a host with epithelial features by c-MYC and the zinc finger protein GKLF. Cell Growth Differ. 10, 423–434 (1999).
Foster, K. W. et al. Induction of KLF4 in basal keratinocytes blocks the proliferation-differentiation switch and initiates squamous epithelial dysplasia. Oncogene 24, 1491–1500 (2005).
Bos, J. L. Ras oncogenes in human cancer: a review. Cancer Res. 49, 4682–4689 (1989).
Palmero, I., Pantoja, C. & Serrano, M. p19ARF links the tumour suppressor p53 to Ras. Nature 395, 125–126 (1998).
Serrano, M., Lin, A. W., McCurrach, M. E., Beach, D. & Lowe, S. W. Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602 (1997).
Peeper, D. S., Dannenberg, J. H., Douma, S., te Riele, H. & Bernards, R. Escape from premature senescence is not sufficient for oncogenic transformation by Ras. Nature Cell Biol. 3, 198–203 (2001).
Sherr, C. J. & McCormick, F. The RB and p53 pathways in cancer. Cancer Cell 2, 103–112 (2002).
Peeper, D. S. et al. A functional screen identifies hDRIL1 as an oncogene that rescues RAS-induced senescence. Nature Cell Biol. 4, 148–153 (2002).
Brummelkamp, T. R., Bernards, R. & Agami, R. Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell 2, 243–247 (2002).
Deng, C., Zhang, P., Harper, J. W., Elledge, S. J. & Leder, P. Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82, 675–684 (1995).
Li, C. et al. The critical role of the PE21 element in oncostatin M-mediated transcriptional repression of the p53 tumor suppressor gene in breast cancer cells. Oncogene 20, 8193–8202 (2001).
Filmus, J. et al. Induction of cyclin D1 overexpression by activated ras. Oncogene 9, 3627–3633 (1994).
Peeper, D. S. et al. Ras signalling linked to the cell-cycle machinery by the retinoblastoma protein. Nature 386, 177–181 (1997).
Sherr, C. J. & Roberts, J. M. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev. 13, 1501–1512 (1999).
Jacobs, J. J. et al. Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19(ARF)) and is amplified in a subset of human breast cancers. Nature Genet. 26, 291–299 (2000).
Land, H., Parada, L. F. & Weinberg, R. A. Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304, 596–602 (1983).
Ruley, H. E. Adenovirus region E1A enables viral and cellular transforming genes to transform primary cells in cultures. Nature 304, 602–606 (1983).
Siegel, P. M. & Massague, J. Cytostatic and apoptotic actions of TGF-β in homeostasis and cancer. Nature Rev. Cancer 3, 807–820 (2003).
Li, C. Y., Suardet, L. & Little, J. B. Potential role of WAF1/Cip1/p21 as a mediator of TGF-β cytoinhibitory effect. J. Biol. Chem. 270, 4971–4974 (1995).
Elbendary, A. et al. Transforming growth factor β1 can induce CIP1/WAF1 expression independent of the p53 pathway in ovarian cancer cells. Cell Growth Differ. 5, 1301–1307 (1994).
Datto, M. B. et al. Transforming growth factor β induces the cyclin-dependent kinase inhibitor p21 through a p53-independent mechanism. Proc. Natl Acad. Sci. USA 92, 5545–5549 (1995).
Zarling, J. M. et al. Oncostatin M: a growth regulator produced by differentiated histiocytic lymphoma cells. Proc. Natl Acad. Sci. USA 83, 9739–9743 (1986).
Bellido, T., O'Brien, C. A., Roberson, P. K. & Manolagas, S. C. Transcriptional activation of the p21(WAF1,CIP1,SDI1) gene by interleukin-6 type cytokines. A prerequisite for their pro-differentiating and anti-apoptotic effects on human osteoblastic cells. J. Biol. Chem. 273, 21137–21144 (1998).
Shiohara, M. et al. Absence of WAF1 mutations in a variety of human malignancies. Blood 84, 3781–3784 (1994).
Rowland, B. D. et al. E2F transcriptional repressor complexes are critical downstream targets of p19(ARF)/p53-induced proliferative arrest. Cancer Cell 2, 55–65 (2002).
Jenkins, T. D., Opitz, O. G., Okano, J. & Rustgi, A. K. Transactivation of the human keratin 4 and Epstein-Barr virus ED-L2 promoters by gut-enriched Kruppel-like factor. J. Biol. Chem. 273, 10747–10754 (1998).
Berns, K. et al. A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 428, 431–437 (2004).
Dirac, A. M. & Bernards, R. Reversal of senescence in mouse fibroblasts through lentiviral suppression of p53. J. Biol. Chem. 278, 11731–11734 (2003).
Raman, V. et al. Compromised HOXA5 function can limit p53 expression in human breast tumours. Nature 405, 974–978 (2000).
Acknowledgements
We acknowledge S. Douma, P. van der Sluis and A. Zwaagstra for technical assistance; A. Dirac, M. Voorhoeve and R. Agami for generously providing p(Retro)SUPER vectors encoding p53, cyclin-D1 and p21CIP1; A. Rustgi for a KLF4 expression vector; M. Oren and S. Sukumar for p53 promoter constructs; E. Roos for the pLIPE vector; M. Eilers for a p53-responsive reporter; and M. Epping for p21CIPI−/− MEFs. We thank A. Berns for critically reading the manuscript. This work was supported by grants from the Dutch Cancer Society (KWF) and The Netherlands organization for scientific research (NWO) to B.D.R. and D.S.P.
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Rowland, B., Bernards, R. & Peeper, D. The KLF4 tumour suppressor is a transcriptional repressor of p53 that acts as a context-dependent oncogene. Nat Cell Biol 7, 1074–1082 (2005). https://doi.org/10.1038/ncb1314
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DOI: https://doi.org/10.1038/ncb1314
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