ERBB2 oncogene; promoter; transcription; breast cancer; BT-474; MDA-MB-231; AP-2 transcription factor; transfection; luciferase; EMSA; chromatin immunoprecipitation, ChIP; DNA binding
Abstract :
[en] Overexpression of the ERBB2 gene occurs in 30% of human breast cancers and is correlated with poor prognosis. The deregulation is the consequence of an increased transcription level and gene amplification. Several laboratories, including our own, have identified, in the proximal promoter, enhancers implicated in the gene overexpression. However, our previous studies of a 6-kb ERBB2 promoter fragment revealed the presence of repressing fragments, which were able to overcome the effect of the proximal enhancers. These repressing elements were functional in all cell lines, regardless of their endogenous ERBB2 expression level. Here, we show that a distal ERBB2 promoter region restores high transcription rates specifically in ERBB2 overexpressing breast cancer cells. This distal promoter region thus contains enhancers essential for the overexpression of the gene. By EMSA, performed with nuclear extract of cells overexpressing (BT-474) or not (MDA-MB-231) the ERBB2 gene, we show that at least two sequences of the distal promoter region are bound exclusively by BT-474 extract. Further experiments reveal that AP-2 transcription factors contribute to this differential binding activity, by binding recognition sequences located 4500 bp and 4000 bp upstream of the transcription start site. These sites are occupied by AP2 in vivo, as demonstrated by ChIP assay. Inactivation of AP-2 proteins in ERBB2 overexpressing cells reduces the distal promoter activity up to 70%, indicating the AP-2 factors are implicated in the strong distal enhancing effect. Moreover, we identified a 54-bp fragment that is bound specifically by BT-474 nuclear extract. Further experiments did not lead to the identification of the protein responsible for this binding. Our results thus highlight the importance of ERBB2 distal promoter region and further implicate AP-2 in ERBB2 overexpression in breast cancer cells.
Delacroix, Laurence ; Université de Liège - ULiège > Département des sciences cliniques > GIGA-R:Immunopath. - Maladies infect. et médec. inter. gén.
Begon, Dominique ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques - Laboratoire d'Oncologie Moléculaire
Chatel, Guillaume; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques - Laboratoire d'Oncologie Moléculaire
Jackers, Pascale ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques - Laboratoire d'Oncologie Moléculaire
Winkler, Rose ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques - Laboratoire d'Oncologie Moléculaire
Language :
English
Title :
Distal ERBB2 promoter fragment displays specific transcriptional and nuclear binding activities in ERBB2 overexpressing breast cancer cells
Alternative titles :
[fr] Un fragment distal du promoteur ERBB2 présente des activités transcriptionnelles et de liaison nucléaire spécifiques dans les cellules de cancer du sein surexprimant ERBB2
Publication date :
September 2005
Journal title :
DNA and Cell Biology
ISSN :
1044-5498
eISSN :
1557-7430
Publisher :
Mary Ann Liebert, Inc., Larchmont, United States - New York
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE] Fonds Léon Fredericq [BE] CAC - Centre anticancéreux près l'Université de Liège asbl [BE] Fédération Belge contre le Cancer Télévie [BE]
AKIYAMA, T., SUDO, C., OGAWARA, H., TOYOSHIMA, K., and YAMAMOTO, T. (1986). The product of the human c-erbB-2 gene: A 185-kilodalton glycoprotein with tyrosine kinase activity. Science 232, 1644-1646.
ALVAREZ, M., RHODES, S.J., and BIDWELL, J.P. (2003). Context-dependent transcription: All politics is local. Gene 313, 43-57.
BENZ, C.C., O'HAGAN, R.C., RICHTER, B., SCOTT, G.K., CHANG, C.H., XIONG, X., CHEW, K., LJUNG, B.M., EDGERTON, S., THOR, A., et al. (1997). HER2/Neu and the Ets transcription activator PEA3 are coordinately upregulated in human breast cancer. Oncogene 15, 1513-1525.
BOSHER, J.M., TOTTY, N.F., HSUAN, J.J., WILLIAMS, T., and HURST, H.C. (1996). A family of AP-2 proteins regulates c-erbB-2 expression in mammary carcinoma. Oncogene 13, 1701-1707.
BOUCHARD, L., LAMARRE, L., TREMBLAY, P.J., and JOLICOEUR, P. (1989). Stochastic appearance of mammary tumors in transgenic mice carrying the MMTV/c-neu oncogene. Cell 57, 931-936.
COBLEIGH, M.A., VOGEL, C.L., TRIPATHY, D., ROBERT, N.J., SCHOLL, S., FEHRENBACHER, L., WOLTER, J.M., PATON, V., SHAK, S., LIEBERMAN, G., et al. (1999). Multinational study of the efficacy and safety of humanized anti-HER2 monoclonal antibody in women who have HER2-overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease. J. Clin. Oncol. 17, 2639-2648.
DECARY, S., DECESSE, J.T., OGRYZKO, V., REED, J.C., NAGUIB-NEVA, I., HAREL-BELLAN, A., and CREMISI, C.E. (2002). The Retinoblastoma protein binds the promoter of survival gene bcl-2 and regulates its transcription in epithelial cells through transcription factor AP2. Mol. Cell. Biol. 22, 7877-7888.
DI FIORE, P.P., PIERCE, J.H., KRAUS, M.H., SEGATTO, O., KING, C.R., and AARONSON, S.A. (1987). erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells. Science 237, 178-182.
D'SOUZA, B., and TAYLOR-PAPADIMITRIOU, J. (1994). Overexpression of ERBB2 in human mammary epithelial cells signals inhibition of transcription of the E-cadherin gene. Proc. Natl. Acad. Sci. USA 91, 7202-7206.
FARALDO, M.L., RODRIGO, I., BEHRENS, J., BIRCHMEIER, W., and CANO, A. (1997). Analysis of the E-cadherin and P-cadherin promoters in murine keratinocyte cell lines from different stages of mouse skin carcinogenesis. Mol. Carcinog. 20, 33-47.
GROOTECLAES, M., PASLEAU, F., DIJKMANS, H., BERZI, P., ALBERT, A., and WINKLER-GOL, R. (1994). The 6-kilobase c-erbB2 promoter contains positive and negative regulatory elements functional in human mammary cell lines. Cancer Res. 54, 4193-4199.
HOLLYWOOD, D.P., and HURST, H.C. (1993). A novel transcription factor, OB2-1, is required for overexpression of the proto-oncogene c-erbB-2 in mammary tumour lines. EMBO J. 12, 2369-2375.
HURST, H.C. (2001). Update on HER-2 as a target for cancer therapy: The ERBB2 promoter and its exploitation for cancer treatment. Breast Cancer Res. 3, 395-398.
KANG, Z., JANNE, O.A., and PALVIMO, J.J. (2004). Coregulator recruitment and histone modifications in transcriptional regulation by the androgen receptor. Mol. Endocrinol. 18, 2633-2648.
KING, C.R., KRAUS, M.H., and AARONSON, S.A. (1985). Amplification of a novel v-erbB-related gene in a human mammary carcinoma. Science 229, 974-976.
KOZMIK, Z., and PACES, V. (1990). Multiple bandshift assay: Rapid identification and cloning of DNA fragments containing specific protein-binding sites. Gene 90, 287-291.
KRAUS, M.H., POPESCU, N.C., AMSBAUGH, S.C., and KING, C.R. (1987). Overexpression of the EGF receptor-related proto-oncogene erbB-2 in human mammary tumor cell lines by different molecular mechanisms. EMBO J. 6, 605-610.
LAKSHMANAN, G., LIEUW, K.H., LIM, K.C., GU, Y., GROSVELD, F., ENGEL, J.D., and KARIS, A. (1999). Localization of distant urogenital system-, central nervous system-, and endocardium-specific transcriptional regulatory elements in the GATA-3 locus. Mol. Cell. Biol. 19, 1558-1568.
MCPHERSON, L.A., BAICHWAL, V.R., and WEIGEL, R.J. (1997). Identification of ERF-1 as a member of the AP2 transcription factor family. Proc. Natl. Acad. Sci. USA 94, 4342-4347.
MOHIBULLAH, N., DONNER, A., IPPOLITO, J.A., and WILLIAMS, T. (1999). SELEX and missing phosphate contact analyses reveal flexibility within the AP-2α protein: DNA binding complex. Nucleic Acids Res. 27, 2760-2769.
NOTTOLI, T., HAGOPIAN-DONALDSON, S., ZHANG, J., PERKINS, A., and WILLIAMS, T. (1998). AP-2-null cells disrupt morphogenesis of the eye, face, and limbs in chimeric mice. Proc. Natl. Acad. Sci. USA 95, 13714-13719.
PASLEAU, F., GROOTECLAES, M., and GOL-WINKLER, R. (1993). Expression of the c-erbB2 gene in the BT474 human mammary tumor cell line: Measurement of c-erbB2 mRNA half-life. Oncogene 8, 849-854.
PRESS, M.F., CORDON-CARDO, C., and SLAMON, D.J. (1990). Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues. Oncogene 5, 953-962.
RAMESH, T.M., ELLIS, A.W., and SPEAR, B.T. (1995). Individual mouse alpha-fetoprotein enhancer elements exhibit different patterns of tissue-specific and hepatic position-dependent activities. Mol. Cell. Biol. 15, 4947-4955.
ROSS, J.S., and FLETCHER, J.A. (1999). HER-2/neu (c-erb-B2) gene and protein in breast cancer. Am. J. Clin. Pathol. 112, S53-S67.
SCHORLE, H., MEIER, P., BUCHERT, M., JAENISCH, R., and MITCHELL, P.J. (1996). Transcription factor AP-2 essential for cranial closure and craniofacial development. Nature 381, 235-238.
SCOTT, G.K., DANIEL, J.C., XIONG, X., MAKI, R.A., KABAT, D., and BENZ, C.C. (1994). Binding of an ETS-related protein within the DNase 1 hypersensitive site of the HER2/neu promoter in human breast cancer cells. J. Biol. Chem. 269, 19848-19858.
SCOTT, G.K., CHANG, C.H., ERNY, K.M., XU, F., FREDERICKS, W.J., RAUSCHER, F.J., 3RD, THOR, A.D., and BENZ, C.C. (2000). Ets regulation of the erbB2 promoter. Oncogene 19, 6490-6502.
SKINNER, A., and HURST, H.C. (1993). Transcriptional regulation of the c-erbB-3 gene in human breast carcinoma cell lines. Oncogene 8, 3393-3401.
SLAMON, D.J., CLARK, G.M., WONG, S.G., LEVIN, W.J., ULLRICH, A., and MCGUIRE, W.L. (1987). Human breast cancer: Correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235, 177-182.
SLAMON, D.J., LEYLAND-JONES, B., SHAK, S., FUCHS, H., PATON, V., BAJAMONDE, A., FLEMING, T., EIERMANN, W., WOLTER, J., PEGRAM, M., et al. (2001). Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N. Engl. J. Med. 344, 783-792.
SUEN, T.C., and HUNG, M.C. (1990). Multiple cis- and trans-acting elements involved in regulation of the neu gene. Mol. Cell. Biol. 10, 6306-6315.
TURNER, B.C., ZHANG, J., GUMBS, A.A., MAHER, M.G., KAPLAN, L., CARTER, D., GLAZER, P.M., HURST, H.C., HAFFTY, B.C., and WILLIAMS, T. (1998). Expression of AP-2 transcription factors in human breast cancer correlates with the regulation of multiple growth factor signalling pathways. Cancer Res. 58, 5466-5472.
VERNIMMEN, D., BEGON, D., SALVADOR, C., GOFFLOT, S., GROOTECLAES, M., and WINKLER, R. (2003a). Identification of HTF (HER2 transcription factor) as an AP-2 (activator protein-2) transcription factor and contribution of the HTF binding site to ERBB2 gene overexpression. Biochem. J. 370, 323-329.
VERNIMMEN, D., GUEDERS, M., PISVIN, S., DELVENNE, P., and WINKLER, R. (2003b). Different mechanisms are implicated in ERBB2 gene overexpression in breast and in other cancers. Br. J. Cancer 89, 899-906.
WHITE, M.R., and HUNG, M.C. (1992). Cloning and characterization of the mouse neu promoter. Oncogene 7, 677-683.
WILLIAMS, T., and TJIAN, R. (1991). Analysis of the DNA-binding and activation properties of the human transcription factor AP-2. Genes Dev. 5, 670-682.
WINGENDER, E., CHEN, X., HEHL, R., KARAS, H., LIEBICH, I., MATYS, V., MEINHARDT, T., PRUSS, M., REUTER, I., and SCHACHERER, F. (2000). TRANSFAC: An integrated system for gene expression regulation. Nucleic Acids Res. 28, 316-319.
YARDEN, Y., and SLIWKOWSKI, M.X. (2001). Untangling the ErbB signalling network. Nat. Rev. Mol. Cell Biol. 2, 127-137.
ZENG, Y.X., SOMASUNDARAM, K., and EL-DEIRY, W.S. (1997). AP2 inhibits cancer cell growth and activates p21WAF1/CIP1 expression. Nat. Genet. 15, 78-82.
ZHAO, F., SATODA, M., LICHT, J.D., HAYASHIZAKI, Y., and 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, 40755-40760.
ZHU, C.H., and DOMANN, F.E. (2002). Dominant negative interference of transcription factor AP-2 causes inhibition of ErbB-3 expression and suppresses malignant cell growth. Breast Cancer Res. Treat. 71, 47-57.
