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Cisplatin resistance in human cervical, ovarian and lung cancer cells

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Abstract

Purpose

This study was performed to determine whether or not in cervical, ovarian and lung cancer cell lines, free radicals (ROS) play a role in cisplatin cytotoxicity and activation of the mitochondrial and JNK/p38 pathways. The role of the enzyme, dihydrodiol dehydrogenase (DDH1), in the activation/deactivation of this pathway and how this may be related to the development of resistance was also investigated.

Methods

Mitochondrial membrane potential and ROS analysis were performed by flow cytometry, P-JNK and P-p38 by western blotting and mRNA by RT-PCR. Dihydrodiol dehydrogenase (DDH1) and thioredoxin knockdowns were prepared by standard techniques.

Results

Cisplatin treatment of a cervical cancer cell line resulted in ROS production with mitochondrial membrane depolarization and phosphorylation of JNK and p38. N-acetyl-cysteine, a free radical scavenger, ameliorated these effects. Treatment of the sensitive cells with H2O2 produced similar effects but at shorter incubation times. Similar results were observed with an ovarian cell line. Downregulation of dihydrodiol dehydrogenase in the cisplatin-resistant cervical and lung cancer cell lines resulted in increased drug sensitivity with detectable production of ROS and activation of the JNK/p38 pathways; however, downregulation of thioredoxin in the cervical cells had minimal effect.

Conclusion

Dihydrodiol dehydrogenase appears to play a role in cisplatin resistance in cervical, ovarian and lung cancer cells which includes mitochondrial membrane depolarization, ROS production and activation of the JNK pathway. However, its mode of action cannot be mimicked by an ROS scavenger so its mechanism of action is more complex (a not unexpected finding considering its role in xenobiotic activation/countering oxidative stress).

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Abbreviations

DDH:

Dihydrodiol dehydrogenase

THR:

Thioredoxin 1

ROS:

Reactive oxygen species

NAC:

N-acetyl-cysteine

H2O2 :

Hydrogen peroxide

DCF:

5-(and 6-)-chloromethyl-2′, 7′-dichlorodihydrofluorescein diacetate, acetyl ester

DHE:

Dihydroethidium

JC1:

5′, 6, 6′-Tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolycarbocyanine iodide

References

  1. Ozols RF, Williams SD (1989) Testicular cancer. Curr Probl Cancer 13:287–335

    Article  Google Scholar 

  2. Loehrer PJ, Einhorn LH (1984) Drugs five years later, Cisplatin. Ann Intern Med 100:704–713

    Article  CAS  PubMed  Google Scholar 

  3. Ferlay J, Shin H, Bray F, Forman D, Mathers C, Parkin D (2008) GLOBOCAN2008: cancer incidence and mortality worldwide: Lyon. International Agency for Research on Cancer, France

    Google Scholar 

  4. Pinto A, Lippard SJ (1985) Binding of the antitumor drug Cis-diamminedichloroplatinum (II) (cisplatin) to DNA. Biochem Biophys Acta 780:167–180

    CAS  PubMed  Google Scholar 

  5. Hector S, Bolanowska-Higdon W, Zdanowicz J, Hitt S, Pendyla L (2001) In vitro studies on the mechanisms of oxaliplatin resistance. Cancer Chemother Pharmacol 48:398–406

    Article  CAS  PubMed  Google Scholar 

  6. Andrews PA, Velury S, Mann SC, Howell SB (1992) Cisdiamminedichloroplatinum (II) accumulation in sensitive and resistant human ovarian cells. Cancer Res 48:68–73

    Google Scholar 

  7. Mamenta EL, Poma EE, Kaufman WK, Delmastro DA, Grady HL, Chaney SG (1994) Enhanced replicative bypass of platinum: DNA adducts in cisplatin resistant human ovarian carcinoma cell lines. Cancer Res 54:3500–3508

    CAS  PubMed  Google Scholar 

  8. Christen RJ, Jekunen AP, Jones JA, Thiebaut F, Shalinsky DR, Howell SB (1993) In Vitro modulation of cisplatin accumulation in human ovarian cells by pharmacological alteration of microtubules. J Clin Invest 92:431–440

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Parekh HK, Simpkins H (1995) The differential expression of cytokeratin 18 in cisplatin-sensitive and resistant human ovarian adenocarcinoma cells and its association with drug sensitivity. Cancer Res 55:5203–5206

    CAS  PubMed  Google Scholar 

  10. Andrews PA, Albright KD (1992) Mitochondrial defects in cis-diamminedichloroplatinum (II)-resistant human ovarian carcinoma cells. Cancer Res 52:1895–1901

    CAS  PubMed  Google Scholar 

  11. Isonishi S, Jekunen AP, Hom DH, Eastman A, Edelstein PS, Thiebaut FB, Christen RD, Howell SB (1992) Modulation of cisplatin sensitivity and growth rate of an ovarian carcinoma cell line by bombesin and tumor necrosis factor. J Clin Invest 90:1436–1442

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Deng HB, Adikari M, Parekh HK, Simpkins H (2002) Increased expression of dihydrodiol dehydrogenase induces resistance to cisplatin in human ovarian carcinoma cells. J Biol Chem 277:15035–15043

    Article  CAS  PubMed  Google Scholar 

  13. Deng HB, Adikari M, Parekh HK, Simpkins H (2004) Ubiquitous induction of resistance to platinum drugs in human ovarian, cervical, germ cell and lung carcinoma tumor cells overexpressing isoforms 1 and 2 of dihydrodriol dehydrogenase. Cancer Chemother Pharmacol 54:301–307

    Article  CAS  PubMed  Google Scholar 

  14. Chen J, Adikari M, Pallai R, Parekh HK, Simpkins H (2008) Dihydrodiol dehydrogenase regulates the generation of reactive oxygen species and the development of cisplatin resistance in human ovarian carcinoma cells. Cancer Chemother Pharmacol 61:979–987

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Hung JJ, Chow KC, Wang HW, Wang LS (2006) Expression of dihydrodiol dehydrogenase and resistance to chemotherapy and radiotherapy in adenocarcinoma cells of lung. Anticancer Res 26:2949–2956

    CAS  PubMed  Google Scholar 

  16. Chen YJ, Yuan GC, Chan KC, Wang PH, Lai CR, Yen MS, Wang LS (2005) Overexpression of dihydrodiol dehydrogenase is associated with cisplatin-based chemotherapy resistance in ovarian cancer patients. Gynecol Oncol 97:110–117

    Article  CAS  PubMed  Google Scholar 

  17. Hsu NY, Ho HC, Chow KC, Lin TY, Shih CS, Wang LS, Tsai CM (2001) Overexpression of dihydrodiol dehydrogenase as a prognostic marker of non-small cell lung cancer. Cancer Res 61:2727–2731

    CAS  PubMed  Google Scholar 

  18. Chen CY, Hsu CP, Hsu NY, Shih CS, Lin TY, Chow KC (2002) Expression of dihydrodiol dehydrogenase in the resected stage I non-small cell lung cancer. Oncol Rep 9:515–519

    CAS  PubMed  Google Scholar 

  19. Kuang P, Zhou C, Li X, Ren S, Li B, Wang Y, Li J, Tang L, Zhang J, Zhao Y (2012) Proteomics-based identification of secreted protein dihydrodiol dehydrogenase 2 as a potential biomarker for predicting cisplatin efficacy in advanced NSCLC patients. Lung Cancer 77:427–432

    Article  PubMed  Google Scholar 

  20. Smithgall TE, Harvey RG, Penning TM (1986) Regio-and stereospecificity of homogeneous 3 alpha hydroxysteroid-dihydrodiol dehydrogenase for transdihydrodiol metabolites of polycyclic aromatic hydrocarbons. J Biol Chem 261:6184–6191

    CAS  PubMed  Google Scholar 

  21. Burczynski ME, Sridhar GR, Palackal NT, Penning TM (2001) The reactive oxygen species-and Michael acceptor-inducible human Aldo-Keto reductase AKR1C1 reduces the α, β-unsaturated aldehyde 4-hydroxy-2-nonenal to 1, 4-Dihydroxy-2-noene. J Biol Chem 276:2890–2897

    Article  CAS  PubMed  Google Scholar 

  22. Pan BF, Nelson JA (2007) Dihydrodiol dehydrogenase in drug resistance and sensitivity of human carcinomas. Cancer Chemother Pharmacol 59:687–702

    Article  Google Scholar 

  23. Takada E, Hata K, Mizuguchi J (2008) C-Jun-NH2-terminal kinase potentiates apoptotic cell death in response to carboplatin in B lymphoma cells. Cancer Chemother Pharmacol 62:569–576

    Article  CAS  PubMed  Google Scholar 

  24. Bragado P, Armesilla A, Silva A, Porras A (2007) Apoptosis by cisplatin requires p53 mediated p38α MAPK activation through ROS generation. Apoptosis 12:1733–1742

    Article  CAS  PubMed  Google Scholar 

  25. Korsch C, Spillman MA, Jackson TA, Jacobsen BM, Murphy SK, Lessey BA, Jordan VC, Bradford AP (2012) DNA profiling analysis of endometrial and ovarian cell lines reveals misidentification redundancy and contamination. Gynecol Oncol 127:241–248

    Article  Google Scholar 

  26. Loetchutinat C, Kothan S, Dechsupa S, Meesungnoen J, Jay-Gerin J, Manketkom S (2005) Spectrofluorometric determination of intracellular levels of reactive oxygen species in drug-sensitive and drug-resistant cancer cells using the 2′,7′-dichlorofluorescein diacetate assay. Radiat Phys Chem 72:323–331

    Article  CAS  Google Scholar 

  27. Camara AKS, Riess ML, Kevin LG, Novalia E, Stowe DL (2004) Hypothermia augments reactive oxygen species detected in the guinea pig isolated perfused heart. Am J Physiol (Heart Circ Physiol) 286: H1289–H1299

  28. Gebauer A, Mirakhur B, Nguyen A, Shue SK, Simpkins H, Dhanasekaran N (2000) Cisplatin resistance involving the defective processing of MEKKI in human ovarian adenocarcinoma 2008/C13 cells. Int J Oncol 16:321–325

    CAS  PubMed  Google Scholar 

  29. Chonghaile TN, Sarosiek KA, Vo TT, Ryan JA, Tammareddi A, Moore VdG, Deng J, Anderson KC, Richardson P, Ta Y-T, Mitsiades CS, Matulonis UA, Drapkin R, Stone R, DeAngelo DJ, McConkey DJ, Sallan SE, Silverman L, Hirsch MS, Carrasco DR, Letai A (2011) Co-treatment mitochondrial priming correlates with clinical response to cytotoxic chemotherapy. Science 334:1129–1133

    Article  PubMed Central  CAS  Google Scholar 

  30. Sodhi A, Gupta P (1986) Increased release of hydrogen peroxide (H2O2) and superoxide anion (O2˙) by murine macrophages in vitro after cisplatin treatment. Int J Immunopharmacol 8:709–714

    Article  CAS  PubMed  Google Scholar 

  31. Kim HS, Hwang JT, Yun H, Chi SG, Lee SJ, Kang I, Yoon KS, Choe WJ, Kim SS, Ha J (2008) Inhibition of AMP-activated protein kinase sensitizes cancer cells to cisplatin-induced apoptosis via hyper-induction of p53. J Biol Chem 283:3731–3742

    Article  CAS  PubMed  Google Scholar 

  32. Biroccio A, Benassi B, Amodei S, Gabellini C, Del Bufalo D, Zupi G (2001) c-Myc down-regulation increases susceptibility to cisplatin through reactive oxygen species-mediated apoptosis in M14 human melanoma cells. Mol Pharmacol 60:174–182

    CAS  PubMed  Google Scholar 

  33. Chen J, Emara N, Solomides C, Parekh H, Simpkins H (2010) Resistance to platinum based chemotherapy in lung cancer cell lines. Cancer Chem Pharm 66:1103–1111

    Article  CAS  Google Scholar 

  34. Burczynski ME, Lin HK, Penning TM (1999) Isoform-specific induction of a human aldo-keto reductase by polycyclic aromatic hydrocarbons (PAHs), electrophiles, and oxidative stress: implications for the alternative pathway of PAH activation catalyzed by human dihydrodiol dehydrogenase. Cancer Res 59:607–614

    CAS  PubMed  Google Scholar 

  35. Yokomizo A, Ono M, Nanri H, Makino Y, Ohga T, Wada M, Okamoto T, Yodol J, Kuwano M, Kohno K (1995) Cellular levels of thioredoxin associated with drug sensitivity to cisplatin, mitomycin C, doxorubicin and ectoposide. Cancer Res 55:4293–4296

    CAS  PubMed  Google Scholar 

  36. Sasada T, Iwata S, Sato N, Kitaoka Y, Hirota K, Nakamura K, Nishiyama A, Taniquchi Y, Takabayashi A, Yodol J (1996) Redox control of resistance to CDDP: protective effect of human thioredoxin against CDDP-induced cytotoxicity. J Clin Invest 97:2268–2276

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Yamada M, Tomida A, Yoshikawa H, Taketani Y, Tsuruo T (1997) Overexpression of thioredoxin does not confer resistance to cisplatin in transfected human ovarian and colon cancer cell lines. Cancer Chemother Pharmacol 40:31–37

    Article  CAS  PubMed  Google Scholar 

  38. Mansouri A, Ridgway LD, Korapati AL, Zhang Q, Tian L, Wang Y, Siddik ZH, Mills GB, Claret FX (2003) Sustained Activation of JNK/p38 MAPK Pathways in Response to Cisplatin Leads to Fas ligand Induction and cell death in ovarian carcinoma cells. J Biol Chem 278:19245–19256

    Article  CAS  PubMed  Google Scholar 

  39. Pallai R, Simpkins H, Chen J, Parekh HK (2010) The CCAAT box binding transcription factor, nuclear factor-Y (NF-Y) regulates transcription of human aldo-keto reductase1C1 (AKR1C1) gene. Gene 459:11–23

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Acknowledgments

The work was supported by the National Institutes of Health [Grant R01-CA098804, Funding Agency: National Cancer Institute] and Staten Island University Hospital.

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Authors and Affiliations

  1. The Feinstein Institute for Medical Research, NS-LIJ Health System, 350 Community Drive, Manhasset, NY, 11030, USA

    Jianli Chen & Henry Simpkins

  2. Department of Pathology and Laboratory Medicine, Staten Island University Hospital, 475 Seaview Avenue, Staten Island, NY, 10305, USA

    Jianli Chen & Henry Simpkins

  3. Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, Thomas Jefferson University, 132 S. 10th Street, 260E Main, Philadelphia, PA, 19107, USA

    Charalambos Solomides

  4. Department of Pathology and Laboratory Medicine, Temple University School of Medicine, 3401 North Broad Street, Philadelphia, PA, 19140, USA

    Hemant Parekh

  5. Jordan Center for Gynecologic Oncology, Penn Perelman Center for Advanced Medicine, 3400 Civic Center Blvd., 3rd floor West, Philadelphia, PA, 19104, USA

    Fiona Simpkins

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  1. Jianli Chen
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Correspondence to Henry Simpkins.

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Chen, J., Solomides, C., Parekh, H. et al. Cisplatin resistance in human cervical, ovarian and lung cancer cells. Cancer Chemother Pharmacol 75, 1217–1227 (2015). https://doi.org/10.1007/s00280-015-2739-2

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  • DOI: https://doi.org/10.1007/s00280-015-2739-2

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