DNA Damage in Healthy Individuals and Respiratory Patients after Treating Whole Blood In vitro with the Bulk and Nano Forms of NSAIDs

Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit COX enzyme activity which affects the inflammatory response. Inflammation is associated with increasing cancer incidence. Pre-clinical and clinical studies have shown that NSAID treatment could cause an anti-tumor effect in cancers. In the present study, blood was taken from healthy individuals (n = 17) and patients with respiratory diseases or lung cancer (n = 36). White blood cells (WBC) were treated with either a micro-suspension, i.e., bulk (B) or nano-suspension (N) of aspirin (ASP) or ibuprofen (IBU) up to 500 μg/ml in the comet assay and up to 125 μg/ml in the micronucleus assay. In this study results were compared against untreated lymphocytes and their corresponding treated groups. The results showed, that NSAIDs in their nano form significantly reduced the DNA damage in WBCs from lung cancer patients in bulk and nano compared to untreated lymphocytes. Also, there was a decrease in the level of DNA damage in the comet assay after treating WBCs from healthy individuals, asthma and COPD groups with aspirin N (ASP N) but not with IBU N. In addition, the number of micronuclei decreased after treatment with NSAIDs in their nano form (ASP N and IBU N) in the healthy as well as in the lung cancer group. However, this was not the case for micronucleus frequency in asthma and COPD patients. These data show that lymphocytes from different groups respond differently to treatment with ASP and IBU as measured by comet assay and micronucleus assay, and that the size of the suspended particles of the drugs affects responses.

[1]  Xiaoyang Xu,et al.  Cancer nanotechnology: the impact of passive and active targeting in the era of modern cancer biology. , 2014, Advanced drug delivery reviews.

[2]  J. Mathers,et al.  Genetics, inheritance and strategies for prevention in populations at high risk of colorectal cancer (CRC). , 2013, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[3]  M. Fenech.  The in vitro micronucleus technique. , 2000, Mutation research.

[4]  J. Potter,et al.  Pre-diagnostic NSAID use but not hormone therapy is associated with improved colorectal cancer survival in women , 2011, British Journal of Cancer.

[5]  Rodney J Scott,et al.  Long-term effect of aspirin on cancer risk in carriers of hereditary colorectal cancer: an analysis from the CAPP2 randomised controlled trial , 2011, The Lancet.

[6]  Y. Kubota,et al.  Analysis of NSAID-Activated Gene 1 Expression in Prostate Cancer , 2010, Urologia Internationalis.

[7]  Randall Harris,et al.  Chemoprevention of lung cancer by non-steroidal anti-inflammatory drugs among cigarette smokers. , 2002, Oncology reports.

[8]  Wenjing Tian,et al.  Meta-analysis on the relationship between nonsteroidal anti-inflammatory drug use and gastric cancer , 2010, European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation.

[9]  Peter York,et al.  Preparation of hydrocortisone nanosuspension through a bottom-up nanoprecipitation technique using microfluidic reactors. , 2009, International journal of pharmaceutics.

[10]  S. Edge,et al.  Genetic variants in COX-2, non-steroidal anti-inflammatory drugs, and breast cancer risk: the Western New York Exposures and Breast Cancer (WEB) Study , 2011, Breast Cancer Research and Treatment.

[11]  Diana Anderson,et al.  Whole-cell Currents Recording from Ion Channels in Human Lymphocytes Treated with Anti-inflammatory Drugs in Nanoparticles Forms , 2014 .

[12]  Diana Anderson,et al.  Sensitivity and specificity of the empirical lymphocyte genome sensitivity (LGS) assay: implications for improving cancer diagnostics , 2014, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[13]  W. Rom,et al.  Chemoprevention of Lung Cancer: Prospects and Disappointments in Human Clinical Trials , 2013, Cancers.

[14]  M. Ashraf,et al.  Non-steroidal anti-inflammatory drugs, tumour immunity and immunotherapy. , 2012, Pharmacological research.

[15]  L. Stark,et al.  c-Src dependency of NSAID-induced effects on NF-κB-mediated apoptosis in colorectal cancer cells. , 2011, Carcinogenesis.

[16]  M. Fenech.  Cytokinesis-block micronucleus cytome assay , 2007, Nature Protocols.

[17]  D. McMillan,et al.  The impact of anti-inflammatory agents on the outcome of patients with colorectal cancer. , 2014, Cancer treatment reviews.

[18]  A. LaCroix,et al.  The Association between NSAID Use and Colorectal Cancer Mortality: Results from the Women's Health Initiative , 2012, Cancer Epidemiology, Biomarkers & Prevention.

[19]  P York,et al.  Comminution of ibuprofen to produce nano-particles for rapid dissolution. , 2011, International journal of pharmaceutics.

[20]  Yongmei Yin,et al.  Meta-analysis on the association between nonsteroidal anti-inflammatory drug use and lung cancer risk. , 2012, Clinical lung cancer.

[21]  C. Ulrich,et al.  IκBKβ and NFκB1, NSAID use and risk of colorectal cancer in the Colon Cancer Family Registry. , 2013, Carcinogenesis.

[22]  Y. S. Shin,et al.  Expression of NSAID-activated gene-1 by EGCG in head and neck cancer: involvement of ATM-dependent p53 expression. , 2013, The Journal of nutritional biochemistry.

[23]  Awadhesh N Jha,et al.  Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals. , 2006, Mutation research.

[24]  T Sofuni,et al.  Report from the In Vitro Micronucleus Assay Working Group. , 2003, Environmental and molecular mutagenesis.

[25]  Diana Anderson,et al.  In vitro sensitivities to UVA of lymphocytes from patients with colon and melanoma cancers and precancerous states in the micronucleus and the Comet assays. , 2012, Mutagenesis.

[26]  M. Fenech,et al.  HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. , 2003, Mutation research.

[27]  M. Fenech,et al.  Report from the In Vitro Micronucleus Assay Working Group , 2003, Mutation research.

[28]  M. Fenech,et al.  Report on the buccal micronucleus assay workshop organized by the International Human Micronucleus (HUMN) project--Antalya, Turkey 2007. , 2008, Mutagenesis.

[29]  D. McMillan,et al.  The impact of aspirin, statins and ACE-inhibitors on the presentation of colorectal neoplasia in a colorectal cancer screening programme , 2013, British Journal of Cancer.

[30]  Amaya Azqueta,et al.  Comet assay to measure DNA repair: approach and applications , 2014, Front. Genet..

[31]  M. Fenech.  Chromosomal biomarkers of genomic instability relevant to cancer. , 2002, Drug discovery today.

[32]  S. Dubinett,et al.  The potential and rationale for COX-2 inhibitors in lung cancer. , 2006, Anti-cancer agents in medicinal chemistry.

[33]  D. Bishop,et al.  Edinburgh Research Explorer Long-term effect of resistant starch on cancer risk in carriers of hereditary colorectal cancer , 2022 .

[34]  N. Cook,et al.  Alternate-Day, Low-Dose Aspirin and Cancer Risk: Long-Term Observational Follow-up of a Randomized Trial , 2013, Annals of Internal Medicine.

[35]  R. Demicheli,et al.  NSAID analgesic ketorolac used perioperatively may suppress early breast cancer relapse: particular relevance to triple negative subgroup , 2012, Breast Cancer Research and Treatment.

[36]  B. Pasche.  Differential effects of aspirin before and after diagnosis of colorectal cancer. , 2013, JAMA.

[37]  E. Giovannucci,et al.  A large cohort study of nonsteroidal anti-inflammatory drugs and renal cell carcinoma incidence in the National Institutes of Health–AARP Diet and Health Study , 2013, Cancer Causes & Control.

[38]  A. Bode,et al.  Naproxen Induces Cell-Cycle Arrest and Apoptosis in Human Urinary Bladder Cancer Cell Lines and Chemically Induced Cancers by Targeting PI3K , 2013, Cancer Prevention Research.

[39]  Edward S. Kim,et al.  Biological Activity of Celecoxib in the Bronchial Epithelium of Current and Former Smokers , 2010, Cancer Prevention Research.

[40]  M D Waters,et al.  IPCS guidelines for the monitoring of genotoxic effects of carcinogens in humans. International Programme on Chemical Safety. , 2000, Mutation research.

[41]  R. Tice,et al.  Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing , 2000, Environmental and molecular mutagenesis.

[42]  M. Kwak,et al.  Induction of p53-independent apoptosis by a novel synthetic hexahydrocannabinol analog is mediated via Sp1-dependent NSAID-activated gene-1 in colon cancer cells. , 2010, Biochemical pharmacology.

[43]  Awadhesh N. Jha,et al.  Comet Assay measurements: a perspective , 2009, Cell Biology and Toxicology.

[44]  J. Baron.  Aspirin and NSAIDs for the prevention of colorectal cancer. , 2009, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.