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Coordinated regulation of two TRAIL-R2/KILLER/DR5 mRNA isoforms by DNA damaging agents, serum and 17β-estradiol in human breast cancer cells

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Abstract

A search of the Genebank database revealed that there are two distinct gene sequences with the common name of TRAIL-R2/Killer/DR5. Using reverse transcription-polymerase chain reaction (RT-PCR), we confirmed the existence of two isoforms of TRAIL-R2/Killer/DR5 mRNA, which we have designated the long and short isoforms based on their electrophoretic mobility. We found that both the long and short mRNA isoforms are ubiquitously expressed in human tissues and cell lines. The long form generally predominates, but the proportion of the two isoforms varies depending on the tissue type. Treatment of MCF-7 human breast cancer cells with the DNA damaging drugs adriamycin, campthothecin, or etoposide causes a coordinated up-regulation of both isoforms. Treatment of the p53-mutant T-47D breast cancer cell line with adriamycin also results in up-regulation of both isoforms, suggesting that adriamycin up-regulates TRAIL-R2/Killer/DR5 expression independent of functional p53. The expression of both mRNA isoforms are increased in MCF-7 cells cultured in charcoal-stripped fetal bovine serum compared to normal serum, suggesting that sex steroid hormones may play a role in the negative regulation of their expression. This was confirmed in MCF-7 cells cultured in stripped serum supplemented with 17β-estradiol, which also resulted in a decrease in the mRNA expression of both isoforms. These results demonstrate that the TRAIL-R2/Killer/DR5 gene gives rise to two distinct forms of mRNA, and that these two forms are coordinately regulated by DNA damage and 17β-estradiol in human breast cancer cells. The functional significance of the two isoforms remains to be determined.

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References

  1. Kerr JFR, Wyllie AH, Currie AR: Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics.Br J Cancer 26: 239–257, 1972

    PubMed  Google Scholar 

  2. Evan G, Littlewood T: A matter of life and cell death. Science 281: 1317–1322, 1998

    PubMed  Google Scholar 

  3. Mikulski SM: Pathogenesis of cancer in view of mutually opposing apoptotic and anti-apoptotic growth signals.Intl J Onc 4: 1257–1263, 1994

    Google Scholar 

  4. Wyllie AH: Apoptosis and carcinogenesis. Euro J Cell Biol 73: 189–197, 1997

    Google Scholar 

  5. Ashkenazi A, Dixit VM: Death receptors: signaling and modulation.Science 281: 1305–1308, 1998

    PubMed  Google Scholar 

  6. Adams JM, Cory S: The Bcl-2 protein family: arbiters of cell survival. Science 281: 1322–1325, 1998

    PubMed  Google Scholar 

  7. Cosman D: A family of ligands for the TNF receptor superfamily.Stem Cells 12: 440–445, 1994

    PubMed  Google Scholar 

  8. Wallach D, Boldin M, Varfolomeev E, Beyaert R, Vandenabeele P, Fiers W: Cell death induction by receptors of the TNF family: towards a molecular understanding. FEBS Lett 410: 96–106, 1997

    PubMed  Google Scholar 

  9. Golstein, P: Cell death: TRAIL and its receptors.Curr Biol 7: R750–R753, 1997

  10. Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA, Goodwin RG: Identification and characterization of a new 15 member of the TNF family that induces apoptosis. Immunity 3: 673–682, 1995

    PubMed  Google Scholar 

  11. Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM: An antagonist decoy receptor and a death domain-containing receptor for TRAIL.Science 277: 815–818, 1997

    PubMed  Google Scholar 

  12. Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A: Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277: 818–821, 1997

    PubMed  Google Scholar 

  13. Sheikh MS, Burns TF, Huang Y, Wu GS, Amundson S, Brooks KS, Fornace AJ Jr, el-Deiry WS: p53-dependent and-independent regulation of the death receptor KILLER/DR5 gene expression in response to genotoxic stress and tumor necrosis factor alpha.Cancer Res 58: 1593–1598, 1998

    PubMed  Google Scholar 

  14. Schneider P, Bodmer JL, Thome M, Hofmann K, Holler N, Tschopp, J: Characterization of two receptors for TRAIL. FEBS Lett 416: 329–334, 1997

    PubMed  Google Scholar 

  15. Chaudhary PM, Eby M, Jasmin A, Bookwalter A, Murray J, Hood L: Death receptor 5, a new member of the TNFR family, and DR4 induce FADD-dependent apoptosis and activate the NF-kappaB pathway.Immunity 7: 821–830, 1997

    PubMed  Google Scholar 

  16. Walczak H, Degli-Esposti MA, Johnson RS, Smolak PJ, Waugh JY, Boiani N, Timour MS, Gerhart MJ, Schooley KA, Smith CA, Goodwin RG and Rauch CT: TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL. EMBO J 16: 5386–5397, 1997

    PubMed  Google Scholar 

  17. Wu GS, Burns TF, McDonald ER III, JiangW, Meng R, Krantz ID, Kao G, Gan DD, Zhou JY, Muschel R, Hamilton SR, Spinner NB, Markowitz S, Wu G, el-Deiry WS: KILLER/DR5 is a DNA damage-inducible p53-regulated death receptor gene.Nat Genet 17: 141–143, 1997

    PubMed  Google Scholar 

  18. MacFarlane M, Ahmad M, Srinivasula SM, Femandes-Alnemri T, Cohen GM, Alnenrri ES: Identification and molecular cloning of two novel receptors for the cytotoxic ligand TRAIL. J Biol Chem 272: 25417–25420, 1997

    PubMed  Google Scholar 

  19. Screaton GR, Mongkolsapaya J, Xu XN, Cowper AE, McMichael AJ, Bell JI: TRICK2, a new alternatively spliced receptor that transduces the cytotoxic signal from TRAIL Curr Biol 9: 693–696, 1997

    Google Scholar 

  20. Wang TT and Phang JM: Effects of estrogen on apoptotic pathways in human breast cancer cell line MCF-7.Cancer Res 55: 2487–2489, 1995

    PubMed  Google Scholar 

  21. Devereux J, Haeberli P and Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12: 387–395, 1984

    PubMed  Google Scholar 

  22. Mowat MR: p53 in tumor progression: life, death, and everything.Adv Cancer Res 74: 25–48, 1998

    PubMed  Google Scholar 

  23. Amellem O, Stokke T, Sandvik JA, Smedshammer L, Pettersen EO: Hypoxia-induced apoptosis in human cells with normal p53 status and function without any alteration in the nuclear protein level. Exp Cell Res 232: 361–370, 1997

    PubMed  Google Scholar 

  24. Schneider P, Thome M, Burns K, Bodmer JL, Hofmann K, Kataoka T, Holler N, Tschopp, J: TRAIL receptors 1 (DR4) and 2 (DR5) signal FADD-dependent apoptosis and activate NF-kappaB.Immunity 7: 831–836, 1997

    PubMed  Google Scholar 

  25. Keane MM, Ettenberg SA, Nau MM, Russell EK, Lipkowitz S: Chemotherapy augments TRAIL-induced apoptosis in breast cell lines. Cancer Res 59: 734–741, 1999

    PubMed  Google Scholar 

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Authors
  1. Thomas T.Y. Wang
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  2. Jiingjau Jeng
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Wang, T.T., Jeng, J. Coordinated regulation of two TRAIL-R2/KILLER/DR5 mRNA isoforms by DNA damaging agents, serum and 17β-estradiol in human breast cancer cells. Breast Cancer Res Treat 61, 87–96 (2000). https://doi.org/10.1023/A:1006432201432

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