Abstract
MCF-7 human breast cancer cells do not express caspase 3, thought by some to be a critical component of the apoptosis cascade. Nonetheless, both mock- and bcl-2-transfected MCF-7 cells undergo apoptosis after treatment with a variety of stimuli, including the DNA-cleaving antimitotic agent, neocarzinostatin (NCS). Transfection with bcl-2 shifts the concentration-response curve to NCS but does not change the phenomenology of apoptosis when it occurs. In both cases, NCS treatment results in condensation and fragmentation of MCF-7 cell nuclei and release of cytochrome c from the mitochondria to the cytosol. This apoptosis is accompanied by decreased levels of Bcl-2 and increased levels of Bax. Using a series of caspase inhibitors with overlapping specificities, enzyme-specific chromogenic substrates, and an antibody specific for activated caspase 7, we have determined that apoptosis in MCF-7 cells proceeds via sequential activation of caspases 9, 7 and 6. P21 is detected only after activation of caspase 7, and P53 is neither expressed at baseline nor up-regulated with apoptosis induction. This pathway bypasses the need for activated caspase 3 in these cells.
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References
Boise LH, Gonzalez-Garcia M, Postema CE, Ding L, Lindsten T, Turka LA, Mao X, Nunez G, Thompson CB . 1993 Cell 74: 597–608
Chandler JM, Cohen GM, MacFarlane M . 1998 J. Biol. Chem. 273: 10815–10818
Cheng EHY, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K, Hardwick JM . 1997 Science 278: 1966–1968
Chinnaiyan AM, Orth K, O'Rourke K, Duan H, Poirier GG, Dixit VM . 1996 J. Biol. Chem. 271: 4573–4576
Cohen GM . 1997 Biochem. J. 326: 1–12
Cortazzo M, Schor NF . 1996 Cancer Res. 56: 1199–1203
Eck-Enriquez K, Kiefer TL, Spriggs LL, Hill SM . 2000 Breast Cancer Res. Treat. 61: 229–239
El-Deiry W, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B . 1993 Cell 75: 817–825
Enari M, Sakahira H, Yokoyoma H, Okawa K, Iwamtsu A, Nagata S . 1998 Nature 391: 43–50
Fraser A, Evan G . 1996 Cell 85: 781–784
Froelich CJ, Dixit VM, Yang X . 1998 Immunol. Today 19: 30–36
Hartsell TL, Yalowich JC, Ritke MK, Martinez AJ, Schor NF . 1995 J. Pharm. Exp. Therap. 275: 479–485
Hartsell TL, Hinman LM, Hamann P, Schor NF . 1996 J. Pharm. Exp. Therap. 277: 1158–1166
Janicke RU, Sprengart ML, Wati MR, Porter AG . 1998 J. Biol. Chem. 273: 9357–9360
Kirsch DG, Doseff A, Chau BN, Lim D, De Souza-Pinto NC, Hansford R, Kastan MB, Lazebnik YA, Hardwick JM . 1999 J. Biol. Chem. 274: 21155–21161
Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD . 1997 Science 275: 1132–1136
Lecoeur H, Gougeon ML . 1996 J. Immunol. Methods 198: 87–99
Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X . 1997 Cell 91: 479–489
Liang Y, Nylander KD, Schor NF . 2001 Pediatr. Res. 49: 211A
Liu X, Kim CN, Yang J, Jemmerson R, Wang X . 1996 Cell 86: 147–157
Liu X, Zou H, Slaughter C, Wang X . 1997 Cell 89: 175–184
Marzo I, Susin SA, Petit PX, Ravagnan L, Brenner C, Larochette N, Zamzami N, Kromer G . 1998 FEBS Lett. 427: 198–202
Medema JP, Toes REM, Scaffidi C, Zheng TS, Flavell RA, Melife CJM, Peter ME, Offring R, Krammer PH . 1997 Eur. J. Immunol. 27: 3492–3498
Merry DE, Korsmeyer SJ . 1997 Annu. Rev. Neurosci. 20: 245–267
Oberhammer F, Wilson JW, Dive C, Morris ID, Hickman JA, Wakeling AE, Walker PR, Sikorska M . 1993 EMBO J. 12: 3679–3684
Okuno S, Shimizu S, Ito T, Nomura M, Hamada E, Tsujimoto Y, Matsuda H . 1998 J. Biol. Chem. 273: 34272–34277
Orth K, Chinnaiyan AM, Garg M, Froelich CJ, Dixit VM . 1996 J. Biol. Chem. 271: 16443–16446
Park J, Jang M, Lee YH, Passaniti A, Kwon TK . 2001 Biochem. Biophys. Res. Commun. 199: 93–98
Parker SB, Eichele G, Zhang P, Rawls A, Sands AT, Bradley A, Olsom EN, Haper JW, Elledge SJ . 1995 Science 267: 1024–1027
Porter AG, Janicke RU . 1999 Cell Death Differ. 6: 99–104
Salvesen GS, Dixit VM . 1997 Cell 91: 443–446
Samali A, Zhivotosky B, Jones D, Nagata S, Orrenius S . 1999 Cell Death Differ. 6: 495–496
Scaffidi CS, Fulda S, Srinivasan A, Friesen C, Li F, Tomaselli KJ, Debatin KM, Krammer PH, Peter ME . 1998 EMBO J. 17: 1675–1687
Schor NF . 1992 J. Clin. Invest. 89: 774–781
Schor NF, Rudin CM, Hartman A-R, Thompson CB, Tyurina YY, Kagan VE . 2000 Oncogene 19: 472–476
Shimizu S, Eguchi Y, Kamiike W, Matsuda H, Tsujimoto Y . 1996 Oncogene 12: 2251–2257
Srinivasula SM, Fernandes-Alnemri T, Zangrillia J, Robertson N, Armstrong RC, Wang L, Trapani JA, Tomaselli KJ, Litwack G, Alnemri ES . 1996 J. Biol. Chem. 271: 27099–27106
Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, Prevost M, Alzari PM, Kroemer G . 1999 J. Exp. Med. 189: 381–393
Susin SA, Zamzami N, Castedo M, Daugas E, Wang HG, Geley S, Fassy F, Reed JC, Kromer G . 1997 J. Exp. Med. 186: 25–37
Talanian RV, Yang X, Turbov J, Seth P, Ghayur T, Casiano CA, Froelich CJ . 1997 J. Exp. Med. 186: 1323–1331
Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X . 1997 Science 275: 1129–1132
Yang X, Sternnick HR, Wang B, Green DR, Janicke RU, Srinivasan A, Srth P, Salvesen GS, Froelich CJ . 1998 J. Biol. Chem. 51: 34278–34283
Zou H, Henzel WJ, Liu A, Lutschg A, Wang X . 1997 Cell 90: 405–413
Acknowledgements
This work was supported by a grant from the National Institutes of Health (R01-CA74289) and the Carol Ann Craumer Endowment Fund of Children's Hospital of Pittsburgh.
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Liang, Y., Yan, C. & Schor, N. Apoptosis in the absence of caspase 3. Oncogene 20, 6570–6578 (2001). https://doi.org/10.1038/sj.onc.1204815
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DOI: https://doi.org/10.1038/sj.onc.1204815