Cell Stress and Chaperones

, Volume 22, Issue 6, pp 811–822 | Cite as

Molecular markers of DNA damage and repair in cervical cancer patients treated with cisplatin neoadjuvant chemotherapy: an exploratory study

  • Nilda E. Real
  • Gisela N. Castro
  • F. Darío Cuello-Carrión
  • Claudia Perinetti
  • Hanna Röhrich
  • Niubys Cayado-Gutiérrez
  • Martin E. Guerrero-Gimenez
  • Daniel R. CioccaEmail author
Original Paper


Neoadjuvant (or induction) chemotherapy can be used for cervical cancer patients with locally advanced disease; this treatment is followed by radical surgery and/or radiation therapy. Cisplatin is considered to be the most active platinum agent drug for this cancer, with a response rate of 20%. In order to understand how the cisplatin treatment affects the stress response, in this work, we performed an exploratory study to analyze a number of stress proteins before and after cisplatin neoadjuvant chemotherapy. The study involved 14 patients; the pre- and post-chemotherapy paired biopsies were examined by hematoxylin and eosin staining and by immunohistochemistry. The proteins evaluated were p53, P16/INK4A, MSH2, nuclear protein transcriptional regulator 1 (NUPR1), and HSPB1 (total: HSPB1/t and phosphorylated: HSPB1/p). These proteins were selected because there is previous evidence of their relationship with drug resistance. The formation of platinum-DNA adducts was also studied. There was a great variation in the expression levels of the mentioned proteins in the pre-chemotherapy biopsies. After chemotherapy, p53 was not significantly affected by cisplatin, as well as P16/INK4A and MSH2 while nuclear NUPR1 content tended to decrease (p = 0.056). Cytoplasmic HSPB1/t expression levels decreased significantly following cisplatin therapy while nuclear HSPB1/t and HSPB1/p tended to increase. Since the most significant changes following chemotherapy appeared in the HSPB1 expression levels, the changes were confirmed by Western blot. The platinum-DNA adducts were observed in HeLa cell in apoptosis; however, in the tumor samples, the platinum-DNA adducts were observed in morphologically healthy tumor cells; these cells displayed nuclear HSPB1/p. Further mechanistic studies should be performed to reveal how HSPB1/p is related with drug resistance. When the correlations of the markers with the response to neoadjuvant chemotherapy were examined, only high pre-chemotherapy levels of cytoplasmic HSPB1/p correlated with a poor clinical and pathological response to neoadjuvant cisplatin chemotherapy (p = 0.056) suggesting that this marker could be useful opening its study in a larger number of cases.


Cervical cancer Neoadjuvant chemotherapy Cisplatin DNA damage Heat shock proteins Molecular markers HSPB1 



This work was partially supported by CONICET grant (PIP 11220110100836 DAS 30844). The authors confirm that the founder had no influence over the study design, content of the article, or selection of this journal.


  1. Al-Mansour Z, Verschraegen C (2010) Locally advanced cervical cancer: what is the standard of care? Curr Opin Oncol 22:503–512CrossRefPubMedGoogle Scholar
  2. Arrossi S (2008) Proyecto para el mejoramiento del Programa Nacional de Prevención del Cáncer de Cuello Uterino en Argentina. 1ra. Edicion. Buenos Aires. Organizacion Panamericana de la Salud, 163 p. ISBN: 978–950–710-114-4Google Scholar
  3. Arrossi S, Maceira V, Paolino M, Sankaranarayanan R (2012) Acceptability and uptake of HPV vaccine in Argentina before its inclusion in the immunization program: a population-based survey. Vaccine 30:2467–2474CrossRefPubMedGoogle Scholar
  4. Cano CE, Hamidi T, Sandi MJ, Iovanna JL (2011) Nupr1: the Swiss-knife of cancer. J Cell Physiol 226:1439–1443CrossRefPubMedGoogle Scholar
  5. Castro GN, Cayado-Gutiérrez N, Zoppino FC, Fanelli MA, Cuello-Carrión FD, Sottile M, Nadin SB, Ciocca DR (2015) Effects of temozolomide (TMZ) on the expression and interaction of heat shock proteins (HSPs) and DNA repair proteins in human malignant glioma cells. Cell Stress Chaperones 20:253–265CrossRefPubMedGoogle Scholar
  6. Cayado-Gutiérrez N, Moncalero VL, Rosales EM, Berón W, Salvatierra EE, Alvarez-Olmedo D, Radrizzani M, Ciocca DR (2013) Downregulation of Hsp27 (HSPB1) in MCF-7 human breast cancer cells induces upregulation of PTEN. Cell Stress Chaperones 18:243–249CrossRefPubMedGoogle Scholar
  7. Chen R, Dai RY, Duan CY, Liu YP, Chen SK, Yan DM, Chen CN, Wei M, Li H (2011) Unfolded protein response suppresses cisplatin-induced apoptosis via autophagy regulation in human hepatocellular carcinoma cells. Folia Biol (Praha) 57:87–95Google Scholar
  8. Chen SF, Nieh S, Jao SW, Liu CL, Wu CH, Chang YC, Yang CY, Lin YS (2012) Quercetin suppresses drug-resistant spheres via the p38 MAPK-Hsp27 apoptotic pathway in oral cancer cells. PLoS One 7(11):e49275CrossRefPubMedPubMedCentralGoogle Scholar
  9. Dasari S, Tchounwou PB (2014) Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 0:364–378CrossRefPubMedCentralGoogle Scholar
  10. Doak SH, Jenkins GJ, Parry EM, Griffiths AP, Baxter JN, Parry JM (2014) Differential expression of the MAD2, BUB1 and HSP27 genes in Barrett's oesophagus-their association with aneuploidy and neoplastic progression. Mutat Res 547:133–144CrossRefGoogle Scholar
  11. Elledge RM, Ciocca DR, Langone G, McGuire WL (1993) Estrogen receptor, progesterone receptor, and HER-2/neu protein in breast cancers from pregnant patients. Cancer 71:2499–2506CrossRefPubMedGoogle Scholar
  12. Fanelli MA, Cuello Carrión FD, Dekker J, Schoemaker J, Ciocca DR (1998) Serological detection of heat shock protein hsp27 in normal and breast cancer patients. Cancer Epidemiol Biomarkers Prev 7:791–795Google Scholar
  13. Ferlay J, Soerjomataram I, Ervik M, et al. (2012) GLOBOCAN 2012 v1. 0, Cancer incidence and mortality worldwide: IARC cancer Base No. 11. Lyon, France: International Agency for Research [on-line].
  14. Fong CW (2016) Platinum anti-cancer drugs: free radical mechanism of Pt-DNA adduct formation and anti-neoplastic effect. Free Radic Biol Med 95:216–229CrossRefPubMedGoogle Scholar
  15. Hamidi T, Algül H, Cano CE, Sandi MJ, Molejon MI, Riemann M, Calvo EL, Lomberk G, Dagorn J-C, Weih F, Urrutia R, Schmid RM, Iovanna JL (2012) Nuclear protein 1 promotes pancreatic cancer development and protects cells from stress by inhibiting apoptosis. J Clin Invest 122:2092–2103CrossRefPubMedPubMedCentralGoogle Scholar
  16. Jennings JC, Murer B, O'Grady A, Hearn LM, Harvey BJ, Kay EW, Thomas W (2015) Differential p16/INK4A cyclin-dependent kinase inhibitor expression correlates with chemotherapy efficacy in a cohort of 88 malignant pleural mesothelioma patients. Br J Cancer 113:69–75CrossRefPubMedPubMedCentralGoogle Scholar
  17. Kang D, Choi HJ, Kang S, Kim SY, Hwang YS, Je S, Han Z, Kim JH, Song JJ (2015) Ratio of phosphorylated HSP27 to nonphosphorylated HSP27 biphasically acts as a determinant of cellular fate in gemcitabine-resistant pancreatic cancer cells. Cell Signal 27:807–817CrossRefPubMedGoogle Scholar
  18. Kase S, Parikh JG, Rao NA (2009) Expression of heat shock protein 27 and alpha-crystallins in human retinoblastoma after chemoreduction. Br J Ophthalmol 93:541–544CrossRefPubMedGoogle Scholar
  19. Katsogiannou M, Andrieu C, Rocchi P (2014) Heat shock protein 27 phosphorylation state is associated with cancer progression. Front Genet 5:346CrossRefPubMedPubMedCentralGoogle Scholar
  20. Langer R, Ott K, Specht K, Becker K, Lordick F, Burian M, Herrmann K, Schrattenholz A, Cahill MA, Schwaiger M, Hofler H, Wester HJ (2008) Protein expression profiling in esophageal adenocarcinoma patients indicates association of heat-shock protein 27 expression and chemotherapy response. Clin Cancer Res 14:8279–8287CrossRefPubMedGoogle Scholar
  21. Li Y, Xiao S, Dan L, Xue M (2015) P16INK4A is required for cisplatin resistance in cervical carcinoma SiHa cells. Oncol Lett 9:1104–1108PubMedGoogle Scholar
  22. Li X, Huang K, Zhang Q, Shen J, Zhou H, Yang R, Wang L, Liu J, Zhang J, Sun H, Jia Y, Du X, Wang H, Deng S, Ding T, Jiang J, Lu Y, Li S, Wang S, Ma D (2016) Early response to neoadjuvant chemotherapy can help predict long-term survival in patients with cervical cancer. Oncotarget. doi:  10.18632/oncotarget.11460
  23. Luvero D, Plotti F, Aloisi A, Capriglione S, Ricciardi R, Miranda A, Lopez S, Scaletta G, De Luca G, Benedetti-Panici P, Angioli R (2016) Patients treated with neoadjuvant chemotherapy + radical surgery + adjuvant chemotherapy in locally advanced cervical cancer: long-term outcomes, survival and prognostic factors in a single-center 10-year follow-up. Med Oncol 33:110CrossRefPubMedGoogle Scholar
  24. Miyazaki T, Kato H, Faried A, Sohda M, Nakajima M, Fukai Y, Masuda N, Manda R, Fukuchi M, Ojima H, Tsukada K, Kuwano H (2005) Predictors of response to chemo-radiotherapy and radiotherapy for esophageal squamous cell carcinoma. Anticancer Res 25:2749–2755PubMedGoogle Scholar
  25. Moureau S, Luessing J, Harte EC, Voisin M, Lowndes NF (2016) A role for the p53 tumour suppressor in regulating the balance between homologous recombination and non-homologous end joining. Open Biol. 6(9)Google Scholar
  26. Nadin S, Ciocca DR (2010) Participation of heat shock proteins in DNA repair mechanisms in cancer. In: DNA Damage Repair, Repair Mechanisms and Aging. Editor: Allison E. Thomas, pp. 165–186. ISBN 978–1–61668-914-8Google Scholar
  27. Nadin SB, Vargas-Roig LM, Drago G, Ibarra J, Ciocca DR (2007) Hsp27, Hsp70 and mismatch repair proteins hMLH1 and hMSH2 expression in peripheral blood lymphocytes from healthy subjects and cancer patients. Cancer Lett 252:131–146CrossRefPubMedGoogle Scholar
  28. Nadin SB, Sottile ML, Montt-Guevara MM, Gauna GV, Daguerre P, Leuzzi M, Gago FE, Ibarra J, Cuello-Carrión FD, Ciocca DR, Vargas-Roig LM (2014) Prognostic implication of HSPA (HSP70) in breast cancer patients treated with neoadjuvant anthracycline-based chemotherapy. Cell Stress Chaperones 19:493–505CrossRefPubMedGoogle Scholar
  29. Narayan S, Sharma N, Kapoor A, Sharma R, Kumar N, Singhal M, Purohit R, Jakhar SL, Beniwal S, Kumar HS, Sharma A (2016) Pros and cons of adding of neoadjuvant chemotherapy to standard concurrent chemoradiotherapy in cervical cancer: a regional cancer center experience. J Obstet Gynaecol India 66:385–390CrossRefPubMedGoogle Scholar
  30. Nicolay NH, Lopez Perez R, Rühle A, Trinh T, Sisombath S, Weber K-J, Ho AD, Debus J, Saffrich R, Huber PE (2016) Mesenchymal stem cells maintain their defining stem cell characteristics after treatment with cisplatin. Sci Rep 6:20035CrossRefPubMedPubMedCentralGoogle Scholar
  31. Oba M, Yano S, Shuto T, Suico MA, Eguma A, Kai H (2008) IFN-gamma down-regulates Hsp27 and enhances hyperthermia-induced tumor cell death in vitro and tumor suppression in vivo. Int J Oncol 32:1317–1324PubMedGoogle Scholar
  32. Palam LR, Gore J, Craven KE, Wilson JL, Korc M (2015) Integrated stress response is critical for gemcitabine resistance in pancreatic ductal adenocarcinoma. Cell Death Dis 6:e1913CrossRefPubMedPubMedCentralGoogle Scholar
  33. Pearcey R, Brundage M, Drouin P, Jeffrey J, Johnston D, Lukka H, MacLean G, Souhami L, Stuart G, Tu D (2002) Phase III trial comparing radical radiotherapy with and without cisplatin chemotherapy in patients with advanced squamous cell cancer of the cervix. J Clin Oncol 20:966–972CrossRefPubMedGoogle Scholar
  34. Poirier MC, Reed E, Litterst CL, Katz D, Gupta-Burt S (1992) Persistence of platinum-ammine-DNA adducts in gonads and kidneys of rats and multiple tissues from cancer patients. Cancer Res 52:149–153PubMedGoogle Scholar
  35. Puy LA, Lo Castro G, Olcese JE, Lotfi HO, Brandi HR, Ciocca DR (1989) Analysis of a 24-kilodalton (KD) protein in the human uterine cervix during abnormal growth. Cancer 64:1067–1073CrossRefPubMedGoogle Scholar
  36. Robl B, Pauli C, Botter SM, Bode-Lesniewska B, Fuchs B (2015) Prognostic value of tumor suppressors in osteosarcoma before and after neoadjuvant chemotherapy. BMC Cancer 15:379CrossRefPubMedPubMedCentralGoogle Scholar
  37. Sawant A, Kothandapani A, Zhitkovich A, Sobol RW, Patrick SM (2015) Role of mismatch repair proteins in the processing of cisplatin interstrand cross-links. DNA Repair (Amst) 35:126–136CrossRefGoogle Scholar
  38. Song TF, Zhang ZF, Liu L, Yang T, Jiang J, Li P (2009) Small interfering RNA-mediated silencing of heat shock protein 27 (HSP27) increases chemosensitivity to paclitaxel by increasing production of reactive oxygen species in human ovarian cancer cells (HO8910). J Int Med Res 37:1375–1388CrossRefPubMedGoogle Scholar
  39. Sottile ML, Losinno AD, Fanelli MA, Cuello-Carrión FD, Montt-Guevara MM, Vargas-Roig LM, Nadin SB (2015) Hyperthermia effects on Hsp27 and Hsp72 associations with mismatch repair (MMR) proteins and cisplatin toxicity in MMR-deficient/proficient colon cancer cell lines. Int J Hyperth 31:464–475CrossRefGoogle Scholar
  40. Stope MB, Wiegank L, Weiss M, Diesing K, Koensgen D, Burchardt M, Zygmunt M, Mustea A (2016) Drug-induced modulation of heat shock protein HSPB1 in an ovarian cancer cell model. Anticancer Res 36:3321–3327PubMedGoogle Scholar
  41. Taba K, Kuramitsu Y, Ryozawa S, Yoshida K, Tanaka T, Maehara S, Maehara Y, Sakaida I, Nakamura K (2010) Heat-shock protein 27 is phosphorylated in gemcitabine-resistant pancreatic cancer cells. Anticancer Res 30:2539–2543PubMedGoogle Scholar
  42. Tadessse SK (2015) Socio-economic and cultural vulnerabilities to cervical cancer and challenges faced by patients attending care at Tikur Anbessa hospital: a cross sectional and qualitative study. BMC Womens Health 15:75CrossRefGoogle Scholar
  43. Thakur B, Ray O (2016) p53 loses grip on PIK3CA expression leading to enhanced cell survival during platinum resistance. Mol Oncol 10:1283–1295CrossRefPubMedPubMedCentralGoogle Scholar
  44. Vargas-Roig LM, Gago FE, Tello O, Aznar JC, Ciocca DR (1998) Heat shock protein expression and drug resistance in breast cancer patients treated with induction chemotherapy. Int J Cancer 79:468–475Google Scholar
  45. Vincent AJ, Ren S, Harris LG, Devine DJ, Samant RS, Fodstad O, Shevde LA (2012) Cytoplasmic translocation of p21 mediates NUPR1-induced chemoresistance NUPR1 and p21 in chemoresistance. FEBS Lett 586:3429–3434CrossRefPubMedGoogle Scholar
  46. World Health Organization (2016) Human papillomavirus (HPV) and cervical cancer. On line
  47. Xie Q, Liang J, Rao Q, Xie X, Li R, Liu Y, Zhou H, Han J, Yao T, Lin Z (2016) Aldehyde dehydrogenase 1 expression predicts chemoresistance and poor clinical outcomes in patients with locally advanced cervical cancer treated with neoadjuvant chemotherapy prior to radical hysterectomy. Ann Surg Oncol 23:163–170CrossRefPubMedGoogle Scholar
  48. Yamamoto K, Okamoto A, Isonishi S, Ochiai K, Ohtake Y (2001) Heat shock protein 27 was up-regulated in cisplatin resistant human ovarian tumor cell line and associated with the cisplatin resistance. Cancer Lett 168:173–181CrossRefPubMedGoogle Scholar
  49. Zhu H, Wu J, Zhang W, Luo H, Shen Z, Cheng H, Zhu X (2016) PKM2 enhances chemosensitivity to cisplatin through interaction with the mTOR pathway in cervical cancer. Sci Rep 6:30788CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Cell Stress Society International 2017

Authors and Affiliations

  • Nilda E. Real
    • 1
  • Gisela N. Castro
    • 2
  • F. Darío Cuello-Carrión
    • 2
  • Claudia Perinetti
    • 1
  • Hanna Röhrich
    • 3
  • Niubys Cayado-Gutiérrez
    • 2
  • Martin E. Guerrero-Gimenez
    • 2
  • Daniel R. Ciocca
    • 2
    Email author
  1. 1.Oncology DepartmentHospital Diego Paroissien of MaipúMendozaArgentina
  2. 2.Laboratory of OncologyInstitute of Medicine and Experimental Biology of Cuyo (IMBECU), National Scientific and Technical Research Council (CONICET)MendozaArgentina
  3. 3.Freie Universitaet BerlinBerlinGermany

Personalised recommendations