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  • Original Paper
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Androgens target prohibitin to regulate proliferation of prostate cancer cells

Abstract

Proteins involved in the growth response of prostate cancer cells to androgen were investigated by comparing the proteomes of LNCaP cells treated with vehicle or androgen. Whole-cell lysates were separated by two-dimensional PAGE, and HPLC-MS/MS was used to identify androgen-regulated proteins. Prohibitin, a protein with cell-cycle regulatory activity, was shown to be downregulated by 50% following androgen stimulation. Western blot and reverse transcription–PCR experiments confirmed the result and showed that regulation occurs at the level of transcription. To determine the importance of prohibitin in androgen-stimulated growth, we used transient transfection to overexpress the protein and RNA interference to knock down the protein. Subsequent FACS analysis showed that cells with reduced levels of prohibitin showed a slight but reproducible increase in the percentage of population in cell cycle, while cells with increased prohibitin levels showed a clear reduction in the percentage entering cell cycle, following dihydrotestosterone stimulation, when compared to untransfected controls. Confocal microscopy showed localization of prohibitin in the nucleus as well as the mitochondria of LNCaP cells. It therefore seems that the regulation of prohibitin is a vital part of the cellular growth response to androgen stimulation in LNCaPs and prohibitin may have a nuclear regulatory role in cell-cycle progression.

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References

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W and Lipman DJ . (1997). Nucleic Acids Res., 25, 3389–3402.

  • Coates PJ, Jamieson DJ, Smart K, Prescott AR and Hall PA . (1997). Curr. Biol., 7, 607–610.

  • Coates PJ, Nenutil R, McGregor A, Picksley SM, Crouch DH, Hall PA and Wright EG . (2001). Exp. Cell. Res., 265, 262–273.

  • Corbett JM, Wheeler CH, Baker CS, Yacoub MH and Dunn MJ . (1994). Electrophoresis, 15, 1459–1465.

  • Dixit VD, Sridaran R, Edmonsond MA, Taub D and Thompson WE . (2003). Endocrinology, 144, 1496–1505.

  • Fellenberg J, Dechant MJ, Ewerbeck V and Mau H . (2003). Int. J. Cancer, 105, 636–643.

  • Gharahdaghi F, Weinberg CR, Meagher DA, Imai BS and Mische SM . (1999). Electrophoresis, 20, 601–605.

  • Gottlieb B, Trifiro M, Lumbroso R and Pinsky L . (1997). Nucleic Acids Res., 25, 158–162.

  • Grayhack JT, Keeler TC and Kozlowski JM . (1987). Cancer, 60, 589–601.

  • Greenlee RT, Hill-Harmon MB, Murray T and Thun M . (2001). CA Cancer J. Clin., 51, 15–36.

  • Ikonen E, Fiedler K, Parton RG and Simons K . (1995). FEBS Lett., 358, 273–277.

  • Jeno P, Mini T, Moes S, Hintermann E and Horst M . (1995). Anal. Biochem., 224, 75–82.

  • Jupe ER, Liu XT, Kiehlbauch JL, McClung JK and Dell’Orco RT . (1996). Exp. Cell. Res., 224, 128–135.

  • Koivisto P, Kolmer M, Visakorpi T and Kallioniemi OP . (1998). Am. J. Pathol., 152, 1–9.

  • Krauss MR, Collins PJ and Blose SH . (1990). Biotechniques, 8, 218–223.

  • Liu XT, Stewart CA, King RL, Danner DA, Dell’Orco RT and McClung JK . (1994). Biochem. Biophys. Res. Commun., 201, 409–414.

  • Mackey AJ, Haystead TA and Pearson WR . (2002). Mol. Cell Proteomics, 1, 139–147.

  • McClung JK, Danner DB, Stewart DA, Smith JR, Schneider EL, Lumpkin CK, Dell’Orco RT and Nuell MJ . (1989). Biochem. Biophys. Res. Commun., 164, 1316–1322.

  • Nijtmans LG, Artal SM, Grivell LA and Coates PJ . (2002). Cell. Mol. Life Sci., 59, 143–155.

  • Nuell MJ, Stewart DA, Walker L, Friedman V, Wood CM, Owens GA, Smith JR, Schneider EL, Dell’ Orco R, Lumpkin CK, Danner DB and McClung JK . (1991). Mol. Cell. Biol., 11, 1372–1381.

  • Peterziel H, Mink S, Schonert A, Becker M, Klocker H and Cato AC . (1999). Oncogene, 18, 6322–6329.

  • Shevchenko A, Wilm M, Vorm O and Mann M . (1996). Anal. Chem., 68, 850–858.

  • Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S and Boyd MR . (1990). J. Natl. Cancer Inst., 82, 1107–1112.

  • Slade MJ, Smith BM, Sinnett HD, Cross NC and Coombes RC . (1999). J. Clin. Oncol., 17, 870–879.

  • Steglich G, Neupert W and Langer T . (1999). Mol. Cell. Biol., 19, 3435–3442.

  • Thompson WE, Branch A, Whittaker JA, Lyn D, Zilberstein M, Mayo KE and Thomas K . (2001). Endocrinology, 142, 4076–4085.

  • Thompson WE, Powell JM, Whittaker JA, Sridaran R and Thomas KH . (1999). Anat. Rec., 256, 40–48.

  • Vander Heiden MG, Choy JS, VanderWeele DJ, Brace JL, Harris MH, Bauer DE, Prange B, Kron SJ, Thompson CB and Rudin CM . (2002). J. Biol. Chem., 277, 44870–44876.

  • Verrijdt G, Haelens A and Claessens F . (2003). Mol. Genet. Metab., 78, 175–185.

  • Visakorpi T, Hyytinen E, Koivisto P, Tanner M, Keinanen R, Palmberg C, Palotie A, Tammela T, Isola J and Kallioniemi OP . (1995). Nat. Genet., 9, 401–406.

  • Wait R, Gianazza E, Eberini I, Sironi L, Dunn MJ, Gemeiner M and Miller I . (2001). Electrophoresis, 22, 3043–3052.

  • Wait R, Miller I, Eberini I, Cairoli F, Veronesi C, Battocchio M, Gemeiner M and Gianazza E . (2002). Electrophoresis, 23, 3418–3427.

  • Wang S, Fusaro G, Padmanabhan J and Chellappan SP . (2002a). Oncogene, 21, 8388–8396.

  • Wang S, Nath N, Adlam M and Chellappan S . (1999a). Oncogene, 18, 3501–3510.

  • Wang S, Nath N, Fusaro G and Chellappan S . (1999b). Mol. Cell. Biol., 19, 7447–7460.

  • Wang S, Zhang B and Faller DV . (2002b). EMBO J., 21, 3019–3028.

  • Weekes J, Wheeler CH, Yan JX, Weil J, Eschenhagen T, Scholtysik G and Dunn MJ . (1999). Electrophoresis, 20, 898–906.

  • Wilm M, Shevchenko A, Houthaeve T, Breit S, Schweigerer L, Fotsis T and Mann M . (1996). Nature, 379, 466–469.

  • Woodlock TJ, Bethlendy G and Segel GB . (2001). Blood Cells Mol. Dis., 27, 27–34.

  • Yan JX, Wait R, Berkelman T, Harry RA, Westbrook JA, Wheeler CH and Dunn MJ . (2000). Electrophoresis, 21, 3666–3672.

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Acknowledgements

We thank Dr M Slade for the RT–PCR control primers, Dr A Cato for the kind gift of the PC3wtAR cells, Mr G Brooke, Mrs V Patel, Miss S Powell, Dr A Varela-Carver and Dr H Whitaker for advice on the techniques used in this paper and Dr S Ali for helpful discussion and critical reading of the manuscript. We acknowledge the support from the HGMP-RC and IMAGE for providing the prohibitin cDNA clone (number 3010198). This work was funded by The Prostate Cancer Charity, and E Lam was supported by Cancer Research UK.

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Correspondence to Charlotte L Bevan.

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Gamble, S., Odontiadis, M., Waxman, J. et al. Androgens target prohibitin to regulate proliferation of prostate cancer cells. Oncogene 23, 2996–3004 (2004). https://doi.org/10.1038/sj.onc.1207444

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