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Naturally occurring dominant-negative Stat5 suppresses transcriptional activity of estrogen receptors and induces apoptosis in T47D breast cancer cells

Abstract

Signal transducer and activator of transcription (Stat) 5 regulates growth, differentiation, and survival of mammary and hematopoietic cells. The role of Stat5 in breast cancer has not been established, although Stat5 is critical for some hematopoietic malignancies. We detected for the first time that Stat5b is constitutively activated in human breast cancer cell lines, and analysed the role of Stat5 in estrogen receptor(ER)-positive breast cancer cell lines using dominant-negative variants of Stat5. Two distinct carboxyl-truncated Stat5a derivatives were generated. Stat5aΔ740 corresponded to a naturally occurring alternative splice variant, and Stat5aΔ713 was analogous to an 80 kDa Stat5a product of a nuclear protease. Stat5aΔ740 and Stat5aΔ713 displayed comparable dominant-negative properties and suppressed transcriptional activity of wild-type Stat5a and Stat5b equally well. Cotransfection experiments revealed that Stat5aΔ740 completely blocked transcriptional activity of endogenous estrogen receptor in T47D and MCF7 cells, and of both ERα and ERβ in COS-7 cells. Stat5aΔ740 was selected for adenoviral delivery, and high-efficiency expression of tyrosine phosphorylated Stat5aΔ740 was achieved in infected cells. Adenoviral-mediated Stat5aΔ740 induced apoptosis in T47D cells but not in caspase-3-negative MCF7 cells. The present study indicates that overexpression of a dominant-negative variant of Stat5 suppresses ER transcriptional activity and induces apoptosis in estrogen-responsive breast cancer tissue culture cells.

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Abbreviations

Stat5:

signal transducer and activator of transcription 5

PRL:

prolactin

ER:

estrogen receptor

ERE:

estrogen-responsive element

E2:

l7β-estradiol

PAGE:

polyacrylamide gel electrophoresis

EMSA:

electrophoretic mobility shift assay

PR:

progesterone receptor

References

  • Beuvink I, Hess D, Flotow H, Hofsteenge J, Groner B and Hynes NE . (2000). J. Biol. Chem., 275, 10247–10255.

  • Bittorf T, Seiler J, Ludtke B, Buchse T, Jaster R and Brock J . (2000). Cell Signal, 12, 23–30.

  • Bjornstrom L, Kilic E, Norman M, Parker MG and Sjoberg M . (2001). J. Mol. Endocrinol., 27, 93–106.

  • Bromberg J and Darnell Jr JE . (2000). Oncogene, 19, 2468–2473.

  • Buettner R, Mora LB and Jove R . (2002). Clin. Cancer Res., 8, 945–954.

  • Cowley SM and Parker MG . (1999). J. Steroid Biochem. Mol. Biol., 69, 165–175.

  • DaSilva L, Rui H, Erwin RA, Howard OM, Kirken RA, Malabarba MG, Hackett RH, Larner AC and Farrar WL . (1996). Mol. Cell Endocrinol., 117, 131–140.

  • Dong F, Liu X, de Koning JP, Touw IP, Henninghausen L, Larner A and Grimley PM . (1998). J. Immunol., 161, 6503–6509.

  • Doppler W, Windegger M, Soratroi C, Tomasi J, Lechner J, Rusconi S, Cato AC, Almlof T, Liden J, Okret S, Gustafsson JA, Richard-Foy H, Starr DB, Klocker H, Edwards D and Geymayer S . (2001). Mol. Cell. Biol., 21, 3266–3279.

  • Faulds Q1MH, Pettersson K, Gustafsson JA and Haldosen LA . (2001). Mol. Endocrinol., 15, 1929–1940.

  • Favre-Young H, Dif F, Roussille F, Demeneix BA, Kelly PA, Edery M and de Luze A . (2000). Mol. Endocrinol., 14, 1411–1424.

  • Frank DA, Mahajan S and Ritz J . (1997). J. Clin. Invest., 100, 3140–3148.

  • Frasor J, Park K, Byers M, Telleria C, Kitamura T, Yu-Lee LY, Djiane J, Park-Sarge OK and Gibori G . (2001). Mol. Endocrinol., 15, 2172–2181.

  • Grimley PM, Dong F and Rui H . (1999). Cytokine Growth Factor Rev., 10, 131–157.

  • Hennighausen L, Robinson GW, Wagner KU and Liu W . (1997). J. Biol. Chem., 272, 7567–7569.

  • Ihle JN . (1996). Cell, 84, 331–334.

  • Janicke RU, Ng P, Sprengart ML and Porter AG . (1998). J. Biol. Chem., 273, 15540–15545.

  • Jones N and Shenk T . (1979). Cell, 17, 683–689.

  • Kazansky AV, Raught B, Lindsey SM, Wang YF and Rosen JM . (1995). Mol. Endocrinol., 9, 1598–1609.

  • Kirken RA, Malabarba MG, Xu J, DaSilva L, Erwin RA, Liu X, Hennighausen L, Rui H and Farrar WL . (1997a). J. Biol. Chem., 272, 15459–15465.

  • Kirken RA, Malabarba MG, Xu J, Liu X, Farrar WL, Hennighausen L, Larner AC, Grimley PM and Rui H . (1997b). J. Biol. Chem., 272, 14098–14103.

  • Lacronique V, Boureux A, Valle VD, Poirel H, Quang CT, Mauchauffe M, Berthou C, Lessard M, Berger R, Ghysdael J and Bernard OA . (1997). Science, 278, 1309–1312.

  • Liu X, Robinson GW, Gouilleux F, Groner B and Hennighausen L . (1995). Proc. Natl. Acad. Sci. USA, 92, 8831–8835.

  • Matsumura I, Ishikawa J, Nakajima K, Oritani K, Tomiyama Y, Miyagawa J, Kato T, Miyazaki H, Matsuzawa Y and Kanakura Y . (1997). Mol. Cell. Biol., 17, 2933–2943.

  • Matsumura I, Kitamura T, Wakao H, Tanaka H, Hashimoto K, Albanese C, Downward J, Pestell RG and Kanakura Y . (1999). EMBO J., 18, 1367–1377.

  • Meyer J, Jucker M, Ostertag W and Stocking C . (1998). Blood, 91, 1901–1908.

  • Moriggl R, Gouilleux-Gruart V, Jahne R, Berchtold S, Gartmann C, Liu X, Hennighausen L, Sotiropoulos A, Groner B and Gouilleux F . (1996). Mol. Cell. Biol., 16, 5691–5700.

  • Mui AL, Wakao H, Kinoshita T, Kitamura T and Miyajima A . (1996). EMBO J., 15, 2425–2433.

  • Nieborowska-Skorska M, Wasik MA, Slupianek A, Salomoni P, Kitamura T, Calabretta B and Skorski T . (1999). J. Exp. Med., 189, 1229–1242.

  • Park SH, Yamashita H, Rui H and Waxman DJ . (2001). Mol. Endocrinol., 15, 2157–2171.

  • Petersen H and Haldosen LA . (1998). Exp. Cell. Res., 243, 347–358.

  • Rosa Santos SC, Dumon S, Mayeux P, Gisselbrecht S and Gouilleux F . (2000). Oncogene, 19, 1164–1172.

  • Saji S, Jensen EV, Nilsson S, Rylander T, Warner M and Gustafsson JA . (2000). Proc. Natl. Acad. Sci. USA, 97, 337–342.

  • Schaber JD, Fang H, Xu J, Grimley PM and Rui H . (1998). Cancer Res., 58, 1914–1919.

  • Socolovsky M, Fallon AE, Wang S, Brugnara C and Lodish HF . (1999). Cell, 98, 181–191.

  • Sonoyama J, Matsumura I, Ezoe S, Satoh Y, Zhang X, Kataoka Y, Takai E, Mizuki M, Machii T, Wakao H and Kanakura Y . (2002). J. Biol. Chem., 277, 8076–8082.

  • Stocklin E, Wissler M, Gouilleux F and Groner B . (1996). Nature, 383, 726–728.

  • Stoecklin E, Wissler M, Schaetzle D, Pfitzner E and Groner B . (1999). J. Steroid Biochem. Mol. Biol., 69, 195–204.

  • Wakao H, Gouilleux F and Groner B . (1994). EMBO J., 13, 2182–2191.

  • Wang D, Stravopodis D, Teglund S, Kitazawa J and Ihle JN . (1996). Mol. Cell. Biol., 16, 6141–6148.

  • Watson CJ . (2001). J. Mammary Gland Biol. Neoplasia, 6, 115–127.

  • Watson CJ and Miller WR . (1995). Br. J. Cancer, 71, 840–844.

  • Wen Z, Zhong Z and Darnell Jr JE . (1995). Cell, 82, 241–250.

  • Yamashita H, Nevalainen MT, Xu J, LeBaron MJ, Wagner KU, Erwin RA, Harmon JM, Hennighausen L, Kirken RA and Rui H . (2001). Mol. Cell. Endocrinol., 183, 151–163.

  • Yamashita H, Xu J, Erwin RA, Farrar WL, Kirken RA and Rui H . (1998). J. Biol. Chem., 273, 30218–30224.

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Acknowledgements

We thank Dr Hallgeir Rui for support in the generation and testing of the dominant-negative Stat5 constructs. This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture in Japan 13671248.

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Correspondence to Hiroko Yamashita.

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Yamashita, H., Iwase, H., Toyama, T. et al. Naturally occurring dominant-negative Stat5 suppresses transcriptional activity of estrogen receptors and induces apoptosis in T47D breast cancer cells. Oncogene 22, 1638–1652 (2003). https://doi.org/10.1038/sj.onc.1206277

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