DNA crosslinks, DNA damage and repair in peripheral blood lymphocytes of non-small cell lung cancer patients treated with platinum derivatives

Fikrova,Petra; Stetina,Rudolf; Hrnciarik,Michal; Hrnciarikova,Dana; Hronek,Miloslav; Zadak,Zdenek

doi:10.3892/or.2013.2805

DNA crosslinks, DNA damage and repair in peripheral blood lymphocytes of non-small cell lung cancer patients treated with platinum derivatives

Authors:
- Petra Fikrova
- Rudolf Stetina
- Michal Hrnciarik
- Dana Hrnciarikova
- Miloslav Hronek
- Zdenek Zadak
View Affiliations

Affiliations: Department of Biological and Medical sciences, Faculty of Pharmacy, Charles University in Prague, Hradec Králové, Czech Republic, Department of Research and Development, University Hospital Hradec Králové, Hradec Králové, Czech Republic, Pulmonary Department, University Hospital Hradec Králové, Hradec Králové, Czech Republic
Published online on: October 23, 2013 https://doi.org/10.3892/or.2013.2805
Pages: 391-396

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Lung cancer is the leading cause of cancer-related mortality in the world. Chemotherapy has been the mainstay of treatment for advanced non-small cell lung cancer (NSCLC) and platinum-based derivatives have been shown to improve overall survival. The aim of the present study was to investigate the DNA damage [single strand breaks (SSBs) and DNA crosslinks] and DNA repair in peripheral blood lymphocytes in patients with NSCLC treated with platinum derivatives using modified comet assay. Twenty patients in the final (4th) stage of NSCLC and 10 age-corresponding healthy controls participated in the study. Alkaline comet assay was performed according to the appropriate protocol. The DNA base excision repair (BER) activity of the controls was significantly higher compared to that of cancer patients, and the activity of DNA nucleotide excision repair (NER) was almost at the same level both in controls and patients. We observed changes in the amount of SSBs and DNA crosslinks during the course of chemotherapy. We found a significantly higher level of SSBs immediately after administration of chemotherapy. Similarly, we found the highest incidence of DNA crosslinks immediately or 1 day after chemotherapy (compared to measurement before chemotherapy). Moreover, we compared the levels of DNA repair in patients who survived chemotherapy with those in patients who died in the course of chemotherapy: the activity of BER was higher in the case of surviving patients, while the levels of NER were essentially the same. The data arising from the present study confirm the findings of other studies dealing with DNA damage and repair in cancer patients treated with chemotherapy. Moreover, our results indicated that despite the fact that cisplatin-DNA adducts are removed by the NER pathway, BER may also play a role in the clinical status of patients and their survival.

View Figures

View References

1	Crohns M, Liippo K, Erhola M, Kankaanranta H, Moilanen E, Alho H and Kellokumpu-Lehtinen P: Concurrent decline of several antioxidants and markers of oxidative stress during combination chemotherapy for small cell lung cancer. Clin Biochem. 12:1236–1245. 2009. View Article : Google Scholar : PubMed/NCBI
2	Gurubhagavatula S, Liu G, Park S, Zhou W, Su L, Wain JC, Lynch TJ, Neuberg DS, David C and Christiani DC: XPD and XRCC1 genetic polymorphisms are prognostic factors in advanced non-small-cell lung cancer patients treated with platinum chemotherapy. J Clin Oncol. 22:2594–2601. 2004. View Article : Google Scholar : PubMed/NCBI
3	Almeida GM, Duarte TL, Steward WP and Jones GD: Detection of oxaliplatin-induced DNA crosslinks in vitro and in cancer patients using the alkaline comet assay. DNA Repair. 5:219–225. 2005. View Article : Google Scholar : PubMed/NCBI
4	Rosenberg B: Fundamental studies with cisplatin. Cancer. 55:2303–2316. 1985. View Article : Google Scholar
5	Lebwohl D and Canetta R: Clinical development of platinum complexes in cancer therapy: an historical perspective and an update. Eur J Cancer. 34:1522–1534. 1998. View Article : Google Scholar : PubMed/NCBI
6	Dronkert ML and Kannar R: Repair of DNA interstrand cross-links. Mutat Res. 486:217–247. 2001. View Article : Google Scholar : PubMed/NCBI
7	Unger FT, Klasen HA, Tchartchian G, de Wilde RL and Witte I: DNA damage induced by cis- and carboplatin as indicator for in vitro sensitivity of ovarian carcinoma cells. BMC Cancer. 9:3592009. View Article : Google Scholar : PubMed/NCBI
8	Rabik CA and Dolan ME: Molecular mechanisms of resistance and toxicity associated with platinating agents. Cancer Treat Rev. 33:9–23. 2007. View Article : Google Scholar : PubMed/NCBI
9	McKenna DJ, McKeown SR and McKelvey-Martin VJ: Potential use of the comet assay in the clinical management of cancer. Mutagenesis. 23:183–190. 2008. View Article : Google Scholar : PubMed/NCBI
10	Faust F, Kassie F, Knasmuller S, Boedecker RH, Mann M and Mersch-Sundermann V: The use of the alkaline comet assay with lymphocytes in human biomonitoring studies. Mutat Res. 566:209–229. 2004. View Article : Google Scholar : PubMed/NCBI
11	Collins AR and Ferguson LR: DNA repair as a biomarker. Mutat Res. 736:2–4. 2012. View Article : Google Scholar : PubMed/NCBI
12	Singh NP, McCoy MT, Tice RR and Schneider EL: A simple technique for the quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 175:184–191. 1988. View Article : Google Scholar : PubMed/NCBI
13	Collins AR: The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol. 26:249–261. 2004. View Article : Google Scholar : PubMed/NCBI
14	Boyum A: Separation of white blood cells. Nature. 204:793–794. 1964. View Article : Google Scholar : PubMed/NCBI
15	Collins AR, Dobson VL, Dusinská M, Kennedy G and Stětina R: The comet assay: what can it really tell us? Mutat Res. 375:183–193. 1997. View Article : Google Scholar : PubMed/NCBI
16	Fikrova P, Stetina R, Hrnciarik M, Rehacek V, Jost P, Hronek M and Zadak Z: Detection of DNA crosslinks in peripheral lymphocytes isolated from patients treated with platinum derivates using modified comet assay. Neoplasma. 60:413–418. 2013. View Article : Google Scholar
17	Collins AR, Dušinská M, Horváthová E, Munro E, Savio M and Stětina R: Inter-individual differences in repair of DNA base oxidation, measured in vitro with the comet assay. Mutagenesis. 16:297–301. 2001. View Article : Google Scholar : PubMed/NCBI
18	Gaivão I, Piasek A, Brevik A, Shaposhnikov S and Collins AR: Comet assay based methods for measuring DNA repair in vitro; estimates of inter- and intraindividual variation. Cell Biol Toxicol. 25:45–52. 2009.PubMed/NCBI
19	Langie SA, Knaapen AM, Brauers KJJ, van Berlo D, van Schooten FJ and Godschalk RWL: Development and validation of a modified comet assay to phenotypically assess nucleotide excision repair. Mutagenesis. 21:153–158. 2006. View Article : Google Scholar : PubMed/NCBI
20	Azqueta A, Shaposhnikov S and Collins AR: DNA repair measured by the comet assay. DNA Repair. Kruman I: InTech; Croatia: 2011, View Article : Google Scholar : 2011, Available from: http://www.intechopen.com/books/dna-repair/dna-repair-measured-by-the-comet-assay.
21	Palyvoda O, Polanska J, Wygoda A and Rzeszowska-Wolny J: DNA damage and repair in lymphocytes of normal individuals and cancer patients: studies by the comet assay and micronucleus tests. Acta Biochim Pol. 50:181–190. 2003.PubMed/NCBI
22	Smith TR, Miller MS, Lohman KK, Case LD and Hu JJ: DNA damage and breast cancer risk. Carcinogenesis. 24:883–889. 2003. View Article : Google Scholar : PubMed/NCBI
23	Rajeswari N, Ahuja YR, Malini U, Chandrashekar S, Balakrishna N, Rao KV and Khar A: Risk assessment in first degree female relatives of breast cancer patients using the alkaline comet assay. Carcinogenesis. 21:557–561. 2000. View Article : Google Scholar
24	Sanchez C, Clementi M, Benitez D, Contreras H, Huidobro C and Castellon E: Effect of GnRH analogs on the expression of TrkA and p75 neurotrophin receptors in primary cell cultures from human prostate adenocarcinoma. Prostate. 65:195–202. 2005. View Article : Google Scholar : PubMed/NCBI
25	Lin X, Wood CG, Shao L, Huang M, Yang H, Dinney CP and Wu X: Risk assessment of renal cell carcinoma using alkaline comet assay. Cancer. 110:282–288. 2007. View Article : Google Scholar : PubMed/NCBI
26	Schabath MB, Grossman HB, Delclos GL, Hernandez LM, Day RS, Davis BR, Lerner SP, Spitz MR and Wu X: Dietary carotenoids and genetic instability modify bladder cancer risk. J Nutr. 134:3362–3369. 2004.PubMed/NCBI
27	Lou J, He J, Zheng W, Jin L, Chen Z, Chen S, Lin Y and Xu S: Investigating the genetic instability in the peripheral lymphocytes of 36 untreated lung cancer patients with comet assay and micronucleus assay. Mutat Res. 617:104–110. 2007. View Article : Google Scholar : PubMed/NCBI
28	Tudek B: Base excision repair modulation as a risk factor for human cancers. Mol Aspects Med. 28:258–275. 2007. View Article : Google Scholar : PubMed/NCBI
29	Paz-Elizur T, Krupsky M, Elinger D, Schechtman E and Livneh Z: Repair of the oxidative DNA damage 8-oxoguanine as a biomarker for lung cancer risk. Cancer Biomark. 1:201–205. 2005.PubMed/NCBI
30	Sánchez-Suárez P, Ostrosky-Wegman P, Gallegos-Hernández F, Peñarroja-Flores R, Toledo-García J, Bravo JL, Del Castillo ER and Benítez-Bribiesca L: DNA damage in peripheral blood lymphocytes in patients during combined chemotherapy for breast cancer. Mutat Res. 640:8–15. 2008.PubMed/NCBI
31	Kopjar N, Garaj-Vrhovac V and Milas I: Assessment of chemotherapy-induced DNA damage in peripheral blood leukocytes of cancer patients using the alkaline comet assay. Teratog Carcinog Mutagen. 22:13–30. 2002. View Article : Google Scholar : PubMed/NCBI
32	Plummer ER, Middleton MR, Jones C, Olsen A, Hickson I, McHugh P, Margison GP, McGown G, Thorncroft M, Watson AJ, Boddy AV, Calvert AH, Harris AL, Newell DR and Curtin NJ: Temozolomide pharmacodynamics in patients with metastatic melanoma: DNA damage and activity of repair enzymes O⁶-alkylguanine alkyltransferase and poly(ADP-ribose) polymerase-1. Clin Cancer Res. 11:3402–3409. 2005. View Article : Google Scholar : PubMed/NCBI
33	Wynne P, Newton C, Ledermann JA, Olaitan A, Mould TA and Hartley JA: Enhanced repair of DNA interstrand crosslinking in ovarian cancer cells from patients following treatment with platinum-based chemotherapy. Br J Cancer. 97:927–933. 2007.PubMed/NCBI
34	Li XQ, Li J, Shi SB, Chen P, Yu LC and Bao QL: Expression of MRP1, BCRP, LRP and ERCC1 as prognostic factors in non-small cell lung cancer patients receiving postoperative cisplatin-based chemotherapy. Int J Biol Markers. 24:230–237. 2009.PubMed/NCBI
35	Rosell R, Lord RV, Taron M and Reguart N: DNA repair and cisplatin resistance in non-small-cell lung cancer. Lung Cancer. 38:217–227. 2002. View Article : Google Scholar : PubMed/NCBI