Advertisement

Induction of p53-mediated senescence is essential for the eventual anticancer therapeutic effect of RH1

  • Joohee Jung
  • Do Young Song
  • Jung Jin Hwang
  • Heon Joo Park
  • Jung Shin Lee
  • Si Yeol Song
  • Seong-Yun JeongEmail author
  • Eun Kyung ChoiEmail author
Research Article
  • 1 Downloads

Abstract

RH1 (2, 5-diaziridinyl-3-(hydroxymethy)-6-methyl-1, 4-benzoquinone) is a bioreductive anticancer drug. The mechanism underlying its therapeutic properties has not yet been elucidated. In this study, we aimed to determine whether RH1 exerts its anticancer effect via p53-mediated apoptosis and senescence in vitro and in vivo. RH1 displayed dose-dependent biphasic effects in vitro, i.e., it induced apoptosis at higher dose and senescence at lower dose accompanied by marked activation of p53. Thus, RH1 primarily induced cell death by apoptosis. The cytotoxicity of RH1 was inhibited in A549 cells treated with the p53-inhibitor pifithrin-α or transfected p53 siRNA and in human colon cancer HCT116 isogenic (p53−/−) cells. At sub-lethal doses of RH1, the cells survived and underwent senescence. The senescent cells showed flattened and enlarged morphology, and exhibited blue color in senescence-associated β-galactosidase staining. These changes were found to be related to p53. RH1-induced senescence decreased in A549-E6 cells (suppressed p53 level) and HCT 116 p53−/− cells. The growth of A549 xenograft tumors in nude mice was significantly delayed by intraperitoneal injection of RH1, and senescent cells were observed in these xenograft tumors. These results suggest that the in vivo anticancer therapeutic effect of RH1 is mediated by senescence via p53 activation.

Keywords

RH1 Anticancer Senescence Apoptosis p53 Tumor growth delay 

Notes

Acknowledgements

This study was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MEST) (NRF-2017R1D1A1B03034359 and NRF-2017R1A2B4008254), Priority Research Centers Program through the NRF funded by MEST (2016R1A6A1A03007648), and a Grant from the Korean Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by Ministry for Health and Welfare, Republic of Korea (HI06C0868 and HI15C0972).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Asher G, Shaul Y (2006) Ubiquitin-independent degradation: lessons from the p53 model. Isr Med Assoc J 8:229–232Google Scholar
  2. Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva N, Vassiliou LV, Kolettas E, Niforou K, Zoumpourlis VC, Takaoka M, Nakagawa H, Tort F, Fugger K, Johansson F, Sehested M, Andersen CL, Dyrskjot L, Orntoft T, Lukas J, Kittas C, Helleday T, Halazonetis TD, Bartek J, Gorgoulis VG (2006) Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature 444:633–637CrossRefGoogle Scholar
  3. Begleiter A, Leith MK, Patel D, Hasinoff BB (2007) Role of NADPH cytochrome P450 reductase in activation of RH1. Cancer Chemother Pharmacol 60:713–723CrossRefGoogle Scholar
  4. Chandeck C, Mooi WJ (2010) Oncogene-induced cellular senescence. Adv Anat Pathol 17:42–48Google Scholar
  5. Chen J, Pan J, Wang J, Song K, Zhu D, Huang C, Duan Y (2016) Soluble egg antigens of Schistosoma japonicum induce senescence in activated hepatic stellate cells by activation of the STAT3/p53/p21 pathway. Sci Rep 6:30957CrossRefGoogle Scholar
  6. Cid-Arregui A (2009) Therapeutic vaccines against human papillomavirus and cervical cancer. Open Virol J 3:67–83CrossRefGoogle Scholar
  7. Collado M, Serrano M (2010) Senescence in tumours: evidence from mice and humans. Nat Rev Cancer 10:51–57CrossRefGoogle Scholar
  8. Danson S, Ward TH, Butler J, Ranson M (2004) DT-diaphorase: a target for new anticancer drugs. Cancer Treat Rev 30:437–449CrossRefGoogle Scholar
  9. Danson S, Johnson T, Ward T, Dawson M, Denneny O, Watson A, Jowle D, Sharpe P, Dive C, Ranson M (2007) Final results of a phase I clinical trial of the bioreductive drug RH1. J Clin Oncol 25:2514Google Scholar
  10. Danson SJ, Johnson P, Ward TH, Dawson M, Denneny O, Dickinson G, Aarons L, Watson A, Jowle D, Cummings J, Robson L, Halbert G, Dive C, Ranson M (2011) Phase I pharmacokinetic and pharmacodynamic study of the bioreductive drug RH1. Ann Oncol 22:1653–1660CrossRefGoogle Scholar
  11. Dehn DL, Inayat-Hussain SH, Ross D (2005) RH1 induces cellular damage in an NAD(P)H:quinone oxidoreductase 1-dependent manner: relationship between DNA cross-linking, cell cycle perturbations, and apoptosis. J Pharmacol Exp Ther 313:771–779CrossRefGoogle Scholar
  12. Digby T, Leith MK, Thliveris JA, Begleiter A (2005) Effect of NQO1 induction on the antitumor activity of RH1 in human tumors in vitro and in vivo. Cancer Chemother Pharmacol 56:307–316CrossRefGoogle Scholar
  13. Ewald JA, Desotelle JA, Wilding G, Jarrard DF (2010) Therapy-induced senescence in cancer. J Natl Cancer Inst 102:1536–1546CrossRefGoogle Scholar
  14. Giaccia AJ, Kastan MB (1998) The complexity of p53 modulation: emerging patterns from divergent signals. Genes Dev 12:2973–2983CrossRefGoogle Scholar
  15. Giard DJ, Aaronson SA, Todaro GJ, Arnstein P, Kersey JH, Dosik H, Parks WP (1973) In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51:1417–1423CrossRefGoogle Scholar
  16. Gong X, Kole L, Iskander K, Jaiswal AK (2007) NRH:quinone oxidoreductase 2 and NAD(P)H:quinone oxidoreductase 1 protect tumor suppressor p53 against 20 s proteasomal degradation leading to stabilization and activation of p53. Cancer Res 67:5380–5388CrossRefGoogle Scholar
  17. Hasinoff BB, Begleiter A (2006) The reductive activation of the antitumor drug RH1 to its semiquinone free radical by NADPH cytochrome P450 reductase and by HCT116 human colon cancer cells. Free Radic Res 40:974–978CrossRefGoogle Scholar
  18. Huang B, Deo D, Xia M, Vassilev LT (2009) Pharmacologic p53 activation blocks cell cycle progression but fails to induce senescence in epithelial cancer cells. Mol Cancer Res 7:1497–1509CrossRefGoogle Scholar
  19. Johnson TM, Meade K, Pathak N, Marques MR, Attardi LD (2008) Knockin mice expressing a chimeric p53 protein reveal mechanistic differences in how p53 triggers apoptosis and senescence. Proc Natl Acad Sci USA 105:1215–1220CrossRefGoogle Scholar
  20. Kepa JK, Ross D (1999) DT-diaphorase activity in NSCLC and SCLC cell lines: a role for fos/jun regulation. Br J Cancer 79:1679–1684CrossRefGoogle Scholar
  21. Lim H, Park BK, Shin SY, Kwon YS, Kim HP (2017) Methyl caffeate and some plant constituents inhibit age-related inflammation: effects on senescence-associated secretory phenotype (SASP) formation. Arch Pharm Res 40:524–535CrossRefGoogle Scholar
  22. Lin AW, Barradas M, Stone JC, Van Aelst L, Serrano M, Lowe SW (1998) Premature senescence involving p53 and p16 is activated in response to constitutive MEK/MAPK mitogenic signaling. Genes Dev 12:3008–3019CrossRefGoogle Scholar
  23. Narita M, Lowe SW (2005) Senescence comes of age. Nat Med 11:920–922CrossRefGoogle Scholar
  24. Park MT, Song MJ, Oh ET, Lee H, Choi BH, Jeong SY, Choi EK, Park HJ (2011) The anti-tumour compound, RH1, causes mitochondria-mediated apoptosis by activating c-Jun N-terminal kinase. Br J Pharmacol 163:567–585CrossRefGoogle Scholar
  25. Shenberger JS, Dixon PS (1999) Oxygen induces S-phase growth arrest and increases p53 and p21(WAF1/CIP1) expression in human bronchial smooth-muscle cells. Am J Respir Cell Mol Biol 21:395–402CrossRefGoogle Scholar
  26. Vazquez A, Bond EE, Levine AJ, Bond GL (2008) The genetics of the p53 pathway, apoptosis and cancer therapy. Nat Rev Drug Discov 7:979–987CrossRefGoogle Scholar
  27. Vilgelm AE, Johnson CA, Prasad N, Yang J, Chen SC, Ayers GD, Pawlikowski JS, Raman D, Sosman JA, Kelley M, Ecsedy JA, Shyr Y, Levy SE, Richmond A (2016) Connecting the dots: therapy-induced senescence and a tumor-suppressive immune microenvironment. J Natl Cancer Inst 108:dvj406CrossRefGoogle Scholar
  28. Winski SL, Hargreaves RH, Butler J, Ross D (1998) A new screening system for NAD(P)H:quinone oxidoreductase (NQO1)-directed antitumor quinones: identification of a new aziridinylbenzoquinone, RH1, as a NQO1-directed antitumor agent. Clin Cancer Res 4:3083–3088Google Scholar
  29. Wyllie FS, Haughton MF, Bond JA, Rowson JM, Jones CJ, Wynford-Thomas D (1996) S phase cell-cycle arrest following DNA damage is independent of the p53/p21(WAF1) signalling pathway. Oncogene 12:1077–1082Google Scholar
  30. Yan C, Kepa JK, Siegel D, Stratford IJ, Ross D (2008) Dissecting the role of multiple reductases in bioactivation and cytotoxicity of the antitumor agent 2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone (RH1). Mol Pharmacol 74:1657–1665CrossRefGoogle Scholar
  31. Yu Y, Gao R, Kaul Z, Li L, Kato Y, Zhang Z, Groden J, Kaul SC, Wadhwa R (2016) Loss-of-function screening to identify miRNAs involved in senescence: tumor suppressor activity of miRNA-335 and its new target CARF. Sci Rep 6:30185CrossRefGoogle Scholar
  32. Zuckerman V, Wolyniec K, Sionov RV, Haupt S, Haupt Y (2009) Tumour suppression by p53: the importance of apoptosis and cellular senescence. J Pathol 219:3–15Google Scholar

Copyright information

© The Pharmaceutical Society of Korea 2019

Authors and Affiliations

  • Joohee Jung
    • 1
    • 2
  • Do Young Song
    • 3
    • 4
  • Jung Jin Hwang
    • 3
    • 4
    • 5
    • 6
  • Heon Joo Park
    • 7
  • Jung Shin Lee
    • 3
    • 8
  • Si Yeol Song
    • 3
    • 6
    • 9
  • Seong-Yun Jeong
    • 3
    • 4
    • 5
    • 6
    Email author
  • Eun Kyung Choi
    • 3
    • 6
    • 9
    Email author
  1. 1.College of PharmacyDuksung Women’s UniversitySeoulKorea
  2. 2.Innovative Drug CenterDuksung Women’s UniversitySeoulKorea
  3. 3.Institute for Innovative Cancer Research, Asan Medical CenterSeoulKorea
  4. 4.Asan Institute for Life Sciences, ASAN Medical CenterUniversity of Ulsan College of MedicineSeoulKorea
  5. 5.Department of Convergence Medicine, ASAN Medical CenterUniversity of Ulsan College of MedicineSeoulKorea
  6. 6.Center for Advancing Cancer Therapeutics, ASAN Medical CenterSeoulKorea
  7. 7.Department of Microbiology, College of MedicineInha UniversityInchonKorea
  8. 8.Department of Internal Medicine, ASAN Medical CenterUniversity of Ulsan College of MedicineSeoulKorea
  9. 9.Department of Radiation Oncology, ASAN Medical CenterUniversity of Ulsan College of MedicineSeoulKorea

Personalised recommendations