Combination of a p53-activating CP-31398 and an MDM2 or a FAK inhibitor produces growth suppressive effects in mesothelioma with wild-type p53 genotype


A majority of mesothelioma had the wild-type p53 genotype but was defective of p53 functions primarily due to a genetic defect in INK4A/ARF region. We examined a growth suppressive activity of CP-31398 which was developed to restore the p53 functions irrespective of the genotype in mesothelioma with wild-type or mutated p53. CP-31398 up-regulated p53 levels in cells with wild-type p53 genotype but induced cell growth suppression in a p53-independent manner. In contrasts, nutlin-3a, an MDM2 inhibitor, increased p53 and p21 levels in mesothelioma with the wild-type p53 genotype and produced growth suppressive effects. We investigated a combinatory effect of CP-31398 and nutlin-2a and found the combination produced synergistic growth inhibition in mesothelioma with the wild-type p53 but not with mutated p53. Western blot analysis showed that the combination increased p53 and the phosphorylation levels greater than treatments with the single agent, augmented cleavages of PARP and caspase-3, and decreased phosphorylated FAK levels. Combination of CP-31398 and defactinib, a FAK inhibitor, also achieved synergistic inhibitory effects and increased p53 with FAK dephosphorylation levels greater than the single treatment. These data indicated that a p53-activating CP-31398 achieved growth inhibitory effects in combination with a MDM2 or a FAK inhibitor and suggested a possible reciprocal pathway between p53 elevation and FAK inactivation.

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Focal adhesion kinase


Half maximal inhibitory concentration


Combination index


Fractions affected


Neurofibromatosis type 2


AMP-activated protein kinase


  1. 1.

    Lee AY, Raz DJ, He B et al (2007) Update on the molecular biology of malignant mesothelioma. Cancer 109:1454–1461

    CAS  Article  Google Scholar 

  2. 2.

    Katzman D, Sterman DH (2018) Updates in the diagnosis and treatment of malignant pleural mesothelioma. Curr Opin Pulm Med 24:319–326

    Article  Google Scholar 

  3. 3.

    Guo G, Chmielecki J, Goparaju C et al (2015) Whole-exome sequencing reveals frequent genetic alterations in BAP1, NF2, CDKN2A, and CUL1 in malignant pleural mesothelioma. Cancer Res 75:264–269

    CAS  Article  Google Scholar 

  4. 4.

    Bueno R, Stawiski EW, Goldstein LD et al (2016) Comprehensive genomic analysis of malignant pleural mesothelioma identifies recurrent mutations, gene fusions and splicing alterations. Nat Genet 48:407–416

    CAS  Article  Google Scholar 

  5. 5.

    Shen J, Cao B, Wang Y et al (2018) Hippo component YAP promotes focal adhesion and tumour aggressiveness via transcriptionally activating THBS1/FAK signalling in breast cancer. J Exp Clin Cancer Res 37:175.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Foster BA, Coffey HA, Morin MJ et al (1999) Pharmacological rescue of mutant p53 conformation and function. Science 286:2507–2510

    CAS  Article  Google Scholar 

  7. 7.

    Tang X, Zhu Y, Han L et al (2007) CP-31398 restores mutant p53 tumor suppressor function and inhibits UVB-induced skin carcinogenesis in mice. J Clin Invest 117:3753–3764

    CAS  Article  Google Scholar 

  8. 8.

    Rippin TM, Bykov VJ, Freund SM et al (2002) Characterization of the p53-rescue drug CP-31398 in vitro and in living cells. Oncogene 21:2119–2129

    CAS  Article  Google Scholar 

  9. 9.

    Takimoto R, Wang W, Dicker DT et al (2002) The mutant p53-conformation modifying drug, CP-31398, can induce apoptosis of human cancer cells and can stabilize wild-type p53 protein. Cancer Biol Ther 1:47–55

    CAS  Article  Google Scholar 

  10. 10.

    Demma MJ, Wong S, Maxwell E et al (2004) CP-31398 restores DNA-binding activity to mutant p53 in vitro but does not affect p53 homologs p63 and p73. J Biol Chem 279:45887–45896

    CAS  Article  Google Scholar 

  11. 11.

    Van Maerken T, Ferdinande L, Taildeman J et al (2009) Antitumor activity of the selective MDM2 antagonist nutlin-3 against chemoresistant neuroblastoma with wild-type p53. J Natl Cancer Inst 101:1562–1574

    Article  Google Scholar 

  12. 12.

    Hopkins-Donaldson S, Belyanskaya LL, Simões-Wüst AP et al (2006) p53-induced apoptosis occurs in the absence of p14(ARF) in malignant pleural mesothelioma. Neoplasia 8:551–559

    CAS  Article  Google Scholar 

  13. 13.

    Shapiro IM, Kolev VN, Vidal CM et al (2014) Merlin deficiency predicts FAK inhibitor sensitivity: a synthetic lethal relationship. Sci Transl Med 6:237–268

    Google Scholar 

  14. 14.

    Soria JC, Gan HK, Blagden SP et al (2016) A phase I, pharmacokinetic and pharmacodynamic study of GSK2256098, a focal adhesion kinase inhibitor, in patients with advanced solid tumors. Ann Oncol 27:2268–2274

    CAS  Article  Google Scholar 

  15. 15.

    Lim ST, Chen XL, Lim Y et al (2008) Nuclear FAK promotes cell proliferation and survival through FERM-enhanced p53 degradation. Mol Cell 29:9–22

    CAS  Article  Google Scholar 

  16. 16.

    Ammoun S, Schmid MC, Zhou L et al (2015) The p53/mouse double minute 2 homolog complex deregulation in merlin-deficient tumours. Mol Oncol 9:236–248

    CAS  Article  Google Scholar 

  17. 17.

    Golubovskaya VM, Finch R, Cance WG (2005) Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J Biol Chem 280:25008–25021

    CAS  Article  Google Scholar 

  18. 18.

    Golubovskaya VM, Finch R, Kweh F et al (2008) p53 regulates FAK expression in human tumor cells. Mol Carcinog 47:373–382

    CAS  Article  Google Scholar 

  19. 19.

    Nakataki E, Yano S, Matsumori Y et al (2006) Novel orthotopic implantation model of human malignant pleural mesothelioma (EHMES-10 cells) highly expressing vascular endothelial growth factor and its receptor. Cancer Sci 97:183–191

    CAS  Article  Google Scholar 

  20. 20.

    Di Marzo D, Forte IM, Indovina P et al (2014) Pharmacological targeting of p53 through RITA is an effective antitumoral strategy for malignant pleural mesothelioma. Cell Cycle 13:652–665

    Article  Google Scholar 

  21. 21.

    Fiorini C, Menegazzi M, Padroni C et al (2013) Autophagy induced by p53-reactivating molecules protects pancreatic cancer cells from apoptosis. Apoptosis 18:337–346

    CAS  Article  Google Scholar 

  22. 22.

    Llanos S, García-Pedrero JM, Morgado-Palacin L et al (2016) Stabilization of p21 by mTORC1/4E-BP1 predicts clinical outcome of head and neck cancers. Nat Commun 7:10438.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Wang W, Rastinejad F, El-Deiry WS (2003) Restoring p53-dependent tumor suppression. Cancer Biol Ther 2:S55–S63

    CAS  PubMed  Google Scholar 

  24. 24.

    Bassett EA, Wang W, Rastinejad F et al (2008) Structural and functional basis for therapeutic modulation of p53 signaling. Clin Cancer Res 14:6376–6386

    CAS  Article  Google Scholar 

  25. 25.

    Chai K, Ning X, Nguyễn TT et al (2018) Heat shock protein 90 inhibitors augmented endogenous wild-type p53 expression but down-regulate the adenovirally-induced expression by inhibiting a proteasome activity. Oncotarget 9:26130–26143

    Article  Google Scholar 

  26. 26.

    Luu Y, Bush J, Cheung KJ Jr et al (2002) The p53 stabilizing compound CP-31398 induces apoptosis by activating the intrinsic Bax/mitochondrial/caspase-9 pathway. Exp Cell Res 276:214–222

    CAS  Article  Google Scholar 

  27. 27.

    Zhong B, Shingyoji M, Hanazono M et al (2019) A p53-stabilizing agent, CP-31398, induces p21 expression with increased G2/M phase through the YY1 transcription factor in esophageal carcinoma defective of the p53 pathway. Am J Cancer Res 9:79–93

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Boudreau HE, Casterline BW, Burke DJ et al (2014) Wild-type and mutant p53 differentially regulate NADPH oxidase 4 in TGF-β-mediated migration of human lung and breast epithelial cells. Br J Cancer 110:2569–2582

    CAS  Article  Google Scholar 

  29. 29.

    Manic G, Obrist F, Sistigu A et al (2015) Trial watch: targeting ATM-CHK2 and ATR-CHK1 pathways for anticancer therapy. Mol Cell Oncol 2:e1012976

    Article  Google Scholar 

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This study was supported by Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science (KAKENHI: 16K09598, 17K10617, 18K15937) and Grant-in-aid from the Nichias Corporation. These funding bodies have not participated in the design of the study, collection, analysis, interpretation of data, or writing of the manuscript.

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Correspondence to Dr. Masatoshi Tagawa.

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The authors declare that there is no conflict of interests in this research. We obtained a grant from Nichias Corporation. It is not a pharmaceutical company but a company making industrial products for building, automobiles and pipes (see The grant is as a kind of their mécénat activities, corporate social contributions, which is aimed to assist for medical research for intractable cancer treatments. We are thereby irrelevant to any employment, consultancy, patents or products in development or marketed products to the company. All the authors agree to publish the data included in the manuscript.

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Zhong, B., Shingyoji, M., Hanazono, M. et al. Combination of a p53-activating CP-31398 and an MDM2 or a FAK inhibitor produces growth suppressive effects in mesothelioma with wild-type p53 genotype. Apoptosis 25, 535–547 (2020).

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  • Mesothelioma
  • CP-31398
  • Nutlin-3a
  • p53
  • FAK