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Research Communications

AP-2α and AP-2γ regulate tumor progression via specific genetic programs

Published Online:stonel.info/10.1096/fj.08-106492

The events occurring during tumor formation and progression display similarities to some of the steps in embryonic morphogenesis. The family of AP-2 proteins consists of five different transcription factors (α, β, γ, δ, and ε) that play relevant roles in embryonic development, as demonstrated by the phenotypes of the corresponding knockout mice. Here, we show that AP-2α and AP-2γ proteins play an essential role in tumorigenesis. Down-modulation of AP-2 expression in tumor cells by RNA interference (RNAi) led to enhanced tumor growth and reduced chemotherapy-induced cell death, as well as migration and invasion. Most of these biological modulations were rescued by AP-2 overexpression. We observed that increased xenotransplant growth was mostly due to highly enhanced proliferation of the tumor cells together with reduced innate immune cell recruitment. Moreover, we showed that migration impairment was mediated, at least in part, by secreted factors. To identify the genetic programs involved in tumorigenesis, we performed whole genome microarray analysis of AP-2α knockdown cells and observed that AP-2α regulates specific genes involved in cell cycle, cell death, adhesion, and migration. In particular, we showed that ESDN, EREG, and CXCL2 play a major role in AP-2 controlled migration, as ablation of any of these genes severely altered migration.—Orso, F., Penna, E., Cimino, D., Astanina, E., Maione, F., Valdembri, D., Giraudo, E., Serini, G., Sismondi, P., De Bortoli, M., Taverna, D. AP-2α and AP-2γ regulate tumor progression via specific genetic programs.


  • 1 Yang, J., Mani, S. A., Donaher, J. L., Ramaswamy, S., Itzykson, R. A., Come, C., Savagner, P., Gitelman, I., Richardson, A., Weinberg, R. A. (2004) Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 117,927-939Crossref Medline
  • 2 Mani, S. A., Yang, J., Brooks, M., Schwaninger, G., Zhou, A., Miura, N., Kutok, J. L., Hartwell, K., Richardson, A. L., Weinberg, R. A. (2007) Mesenchyme forkhead 1 (FOXC2) plays a key role in metastasis and is associated with aggressive basal-like breast cancers. Proc. Natl. Acad. Sci. U. S. A. 104,10069-10074Crossref Medline
  • 3 Hartwell, K. A., Muir, B., Reinhardt, F., Carpenter, A. E., Sgroi, D. C., Weinberg, R. A. (2006) The Spemann organizer gene, Goosecoid, promotes tumor metastasis. Proc. Natl. Acad. Sci. U. S. A. 103,18969-18974Crossref Medline
  • 4 Hilger-Eversheim, K., Moser, M., Schorle, H., Buettner, R. (2000) Regulatory roles of AP-2 transcription factors in vertebrate development, apoptosis and cell-cycle control. Gene 260,1-12Crossref Medline
  • 5 Zhao, F., Satoda, M., Licht, J. D., Hayashizaki, Y., Gelb, B. D. (2001) Cloning and characterization of a novel mouse AP-2 transcription factor, AP-2delta, with unique DNA binding and transactivation properties. J. Biol. Chem. 276,40755-40760Crossref Medline
  • 6 Tummala, R., Romano, R. A., Fuchs, E., Sinha, S. (2003) Molecular cloning and characterization of AP-2 epsilon, a fifth member of the AP-2 family. Gene 321,93-102Crossref Medline
  • 7 Williams, T., Tjian, R. (1991) Characterization of a dimerization motif in AP-2 and its function in heterologous DNA-binding proteins. Science 251,1067-1071Crossref Medline
  • 8 McPherson, L. A., Loktev, A. V., Weigel, R. J. (2002) Tumor suppressor activity of AP2alpha mediated through a direct interaction with p53. J. Biol. Chem. 277,45028-45033Crossref Medline
  • 9 Mertens, P. R., Steinmann, K., Alfonso-Jaume, M. A., En-Nia, A., Sun, Y., Lovett, D. H. (2002) Combinatorial interactions of p53, activating protein-2, and YB-1 with a single enhancer element regulate gelatinase A expression in neoplastic cells. J. Biol. Chem. 277,24875-24882Crossref Medline
  • 10 Modugno, M., Tagliabue, E., Ardini, E., Berno, V., Galmozzi, E., De Bortoli, M., Castronovo, V., Menard, S. (2002) p53-dependent downregulation of metastasis-associated laminin receptor. Oncogene 21,7478-7487Crossref Medline
  • 11 Decary, S., Decesse, J. T., Ogryzko, V., Reed, J. C., Naguibneva, I., Harel-Bellan, A., Cremisi, C. E. (2002) The retinoblastoma protein binds the promoter of the survival gene bcl-2 and regulates its transcription in epithelial cells through transcription factor AP-2. Mol. Cell. Biol. 22,7877-7888Crossref Medline
  • 12 Schorle, H., Meier, P., Buchert, M., Jaenisch, R., Mitchell, P. J. (1996) Transcription factor AP-2 essential for cranial closure and craniofacial development. Nature 381,235-238Crossref Medline
  • 13 Werling, U., Schorle, H. (2002) Conditional inactivation of transcription factor AP-2gamma by using the Cre/loxP recombination system. Genesis 32,127-129Crossref Medline
  • 14 Werling, U., Schorle, H. (2002) Transcription factor gene AP-2 gamma essential for early murine development. Mol. Cell. Biol. 22,3149-3156Crossref Medline
  • 15 Moser, M., Pscherer, A., Roth, C., Becker, J., Mucher, G., Zerres, K., Dixkens, C., Weis, J., Guay-Woodford, L., Buettner, R., Fassler, R. (1997) Enhanced apoptotic cell death of renal epithelial cells in mice lacking transcription factor AP-2beta. Genes Dev. 11,1938-1948Crossref Medline
  • 16 Zeng, Y. X., Somasundaram, K., el-Deiry, W. S. (1997) AP2 inhibits cancer cell growth and activates p21WAF1/CIP1 expression. Nat. Genet. 15,78-82Crossref Medline
  • 17 Wang, D., Shin, T. H., Kudlow, J. E. (1997) Transcription factor AP-2 controls transcription of the human transforming growth factor-alpha gene. J. Biol. Chem. 272,14244-14250Crossref Medline
  • 18 McPherson, L. A., Baichwal, V. R., Weigel, R. J. (1997) Identification of ERF-1 as a member of the AP2 transcription factor family. Proc. Natl. Acad. Sci. U. S. A. 94,4342-4347Crossref Medline
  • 19 Wajapeyee, N., Britto, R., Ravishankar, H. M., Somasundaram, K. (2006) Apoptosis induction by activator protein 2α involves transcriptional repression of Bcl-2. J. Biol. Chem. 281,16207-16219Crossref Medline
  • 20 Huang, S., Jean, D., Luca, M., Tainsky, M. A., Bar-Eli, M. (1998) Loss of AP-2 results in downregulation of c-KIT and enhancement of melanoma tumorigenicity and metastasis. EMBO J. 17,4358-4369Crossref Medline
  • 21 Nyormoi, O., Bar-Eli, M. (2003) Transcriptional regulation of metastasis-related genes in human melanoma. Clin. Exp. Metastasis 20,251-263Crossref Medline
  • 22 Suyama, E., Minoshima, H., Kawasaki, H., Taira, K. (2002) Identification of AP-2-regulated genes by macroarray profiling of gene expression in human A375P melanoma. Nucleic Acids Res. Suppl. ,247-248Medline
  • 23 Gaubatz, S., Imhof, A., Dosch, R., Werner, O., Mitchell, P., Buettner, R., Eilers, M. (1995) Transcriptional activation by Myc is under negative control by the transcription factor AP-2. EMBO J. 14,1508-1519Crossref Medline
  • 24 Gee, J. M., Robertson, J. F., Ellis, I. O., Nicholson, R. I., Hurst, H. C. (1999) Immunohistochemical analysis reveals a tumour suppressor-like role for the transcription factor AP-2 in invasive breast cancer. J. Pathol. 189,514-520Crossref Medline
  • 25 Pellikainen, J., Kataja, V., Ropponen, K., Kellokoski, J., Pietilainen, T., Bohm, J., Eskelinen, M., Kosma, V. M. (2002) Reduced nuclear expression of transcription factor AP-2 associates with aggressive breast cancer. Clin. Cancer Res. 8,3487-3495Medline
  • 26 Karjalainen, J. M., Kellokoski, J. K., Eskelinen, M. J., Alhava, E. M., Kosma, V. M. (1998) Downregulation of transcription factor AP-2 predicts poor survival in stage I cutaneous malignant melanoma. J. Clin. Oncol. 16,3584-3591Crossref Medline
  • 27 Ropponen, K. M., Kellokoski, J. K., Pirinen, R. T., Moisio, K. I., Eskelinen, M. J., Alhava, E. M., Kosma, V. M. (2001) Expression of transcription factor AP-2 in colorectal adenomas and adenocarcinomas; comparison of immunohistochemistry and in situ hybridisation. J. Clin. Pathol. 54,533-538Crossref Medline
  • 28 Ruiz, M., Troncoso, P., Bruns, C., Bar-Eli, M. (2001) Activator protein 2alpha transcription factor expression is associated with luminal differentiation and is lost in prostate cancer. Clin. Cancer Res. 7,4086-4095Medline
  • 29 Bosher, J. M., Williams, T., Hurst, H. C. (1995) The developmentally regulated transcription factor AP-2 is involved in c-erbB-2 overexpression in human mammary carcinoma. Proc. Natl. Acad. Sci. U. S. A. 92,744-747Crossref Medline
  • 30 Gilbertson, R. J., Perry, R. H., Kelly, P. J., Pearson, A. D., Lunec, J. (1997) Prognostic significance of HER2 and HER4 coexpression in childhood medulloblastoma. Cancer Res. 57,3272-3280Medline
  • 31 Bosher, J. M., Totty, N. F., Hsuan, J. J., Williams, T., Hurst, H. C. (1996) A family of AP-2 proteins regulates c-erbB-2 expression in mammary carcinoma. Oncogene 13,1701-1707Medline
  • 32 Pellikainen, J. M., Kosma, V. M. (2007) Activator protein-2 in carcinogenesis with a special reference to breast cancer—a mini review. Intl. J. Cancer 120,2061-2067Crossref Medline
  • 33 Perissi, V., Menini, N., Cottone, E., Capello, D., Sacco, M., Montaldo, F., De Bortoli, M. (2000) AP-2 transcription factors in the regulation of ERBB2 gene transcription by oestrogen. Oncogene 19,280-288Crossref Medline
  • 34 Orso, F., Cottone, E., Hasleton, M. D., Ibbitt, J. C., Sismondi, P., Hurst, H. C., De Bortoli, M. (2004) Activator protein-2gamma (AP-2gamma) expression is specifically induced by oestrogens through binding of the oestrogen receptor to a canonical element within the 5′-untranslated region. Biochem. J. 377,429-438Crossref Medline
  • 35 Ruiz, M., Pettaway, C., Song, R., Stoeltzing, O., Ellis, L., Bar-Eli, M. (2004) Activator protein 2alpha inhibits tumorigenicity and represses vascular endothelial growth factor transcription in prostate cancer cells. Cancer Res. 64,631-638Crossref Medline
  • 36 Jager, R., Friedrichs, N., Heim, I., Buttner, R., Schorle, H. (2005) Dual role of AP-2gamma in ErbB-2-induced mammary tumorigenesis. Breast Cancer Res. Treatment 90,273-280Crossref Medline
  • 37 Wajapeyee, N., Raut, C. G., Somasundaram, K. (2005) Activator protein 2alpha status determines the chemosensitivity of cancer cells: implications in cancer chemotherapy. Cancer Res. 65,8628-8634Crossref Medline
  • 38 Chang, H. Y., Sneddon, J. B., Alizadeh, A. A., Sood, R., West, R. B., Montgomery, K., Chi, J. T., van de Rijn, M., Botstein, D., Brown, P. O. (2004) Gene expression signature of fibroblast serum response predicts human cancer progression: similarities between tumors and wounds. PLoS Biol. 2,E7Crossref Medline
  • 39 Minn, A. J., Gupta, G. P., Siegel, P. M., Bos, P. D., Shu, W., Giri, D. D., Viale, A., Olshen, A. B., Gerald, W. L., Massague, J. (2005) Genes that mediate breast cancer metastasis to lung. Nature 436,518-524Crossref Medline
  • 40 Kobuke, K., Furukawa, Y., Sugai, M., Tanigaki, K., Ohashi, N., Matsumori, A., Sasayama, S., Honjo, T., Tashiro, K. (2001) ESDN, a novel neuropilin-like membrane protein cloned from vascular cells with the longest secretory signal sequence among eukaryotes, is up-regulated after vascular injury. J. Biol. Chem. 276,34105-34114Crossref Medline
  • 41 Nagai, H., Sugito, N., Matsubara, H., Tatematsu, Y., Hida, T., Sekido, Y., Nagino, M., Nimura, Y., Takahashi, T., Osada, H. (2007) CLCP1 interacts with semaphorin 4B and regulates motility of lung cancer cells. Oncogene 26,4025-4031Crossref Medline
  • 42 Pear, W. S., Nolan, G. P., Scott, M. L., Baltimore, D. (1993) Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci. U. S. A. 90,8392-8396Crossref Medline
  • 43 Dull, T., Zufferey, R., Kelly, M., Mandel, R. J., Nguyen, M., Trono, D., Naldini, L. (1998) A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72,8463-8471Medline
  • 44 Chiorino, G., Acquadro, F., Mello Grand, M., Viscomi, S., Segir, R., Gasparini, M., Dotto, P. (2004) Interpretation of expression-profiling results obtained from different platforms and tissue sources: examples using prostate cancer data. Eur. J. Cancer 40,2592-2603Crossref Medline
  • 45 Bookout, A. L., Mangelsdorf, D. J. (2003) Quantitative real-time PCR protocol for analysis of nuclear receptor signaling pathways. Nuclear Recep. Signal. 1,e012[Online]Crossref Medline
  • 46 Kueng, W., Silber, E., Eppenberger, U. (1989) Quantification of cells cultured on 96-well plates. Anal. Biochem. 182,16-19Crossref Medline
  • 47 Hynes, N. E., Taverna, D., Harwerth, I. M., Ciardiello, F., Salomon, D. S., Yamamoto, T., Groner, B. (1990) Epidermal growth factor receptor, but not c-erbB-2, activation prevents lactogenic hormone induction of the beta-casein gene in mouse mammary epithelial cells. Mol. Cell. Biol. 10,4027-4034Crossref Medline
  • 48 Giraudo, E., Inoue, M., Hanahan, D. (2004) An amino-bisphosphonate targets MMP-9-expressing macrophages and angiogenesis to impair cervical carcinogenesis. J. Clin. Invest. 114,623-633Crossref Medline
  • 49 Rasola, A., Geuna, M. (2001) A flow cytometry assay simultaneously detects independent apoptotic parameters. Cytometry 45,151-157Crossref Medline
  • 50 Lazzarato, F., Franceschinis, G., Botta, M., Cordero, F., Calogero, R. A. (2004) RRE: a tool for the extraction of non-coding regions surrounding annotated genes from genomic datasets. Bioinformatics 20,2848-2850Crossref Medline