Association of Tumour Microenvironment with Protein Glycooxidation, DNA Damage, and Nitrosative Stress in Colorectal Cancer

Abstract

Purpose

In this study, we evaluated the total antioxidant capacity, nitrosative stress, and protein/DNA oxidation and glycoxidation products in patients with colorectal cancer regarding histopathological parameters associated with the tumour microenvironment, such as inflammatory infiltration and tumour budding and compare all determined parameters between tumours located in the right and left side of the colon and normal mucosa.

Patients and Methods

Ferric reducing antioxidant power (FRAP), nitrosative stress (myeloperoxidase (MPO), nitrogen oxide (NO), peroxynitrite, and nitrotyrosine), protein oxidation products (protein carbonyls (PC), total thiols, and ischemia modified albumin (IMA)), protein glycooxidation products (tryptophan, kynurenine, N-formylkynurenine, dityrosine, Amadori product, advanced glycation end products (AGE)) and 8-hydroxydeoxyguanosine (8-OHdG) were measured in homogenates from normal and cancerous tissue of 30 patients with colorectal cancer.

Results

Levels of FRAP (p=0.0009), IMA (p=0.0002), kynurenine (p<0.0001), N-formylkynurenine (p<0.0001), dityrosine (p<0.0001), Amadori products (p=0.0024), AGE (p<0.0001), MPO (p<0.0001), NO (p<0.0001) and nitrotyrosine (p=0.0011) were increased, whereas PC (p=0.0004), tryptophan (p<0.0001), 8-OHdG (p<0.0001) and peroxynitrite (p=0.0003) were decreased in the left-side tumour compared to the right-side tumour and normal mucosa.

Conclusion

Our results showed that colorectal cancer is related with disturbances in antioxidant defense and increased oxidative and nitrosative damages to proteins and DNA. These parameters may be useful for evaluation the progression and differentiation of the tumour location. We also demonstrated that redox indicators may depend on the histological type of the tumour and may influence tumour invasion depth, presence of lymph node and distant metastasis, vascular and neural invasion, inflammatory infiltration, and tumour budding, which are part of the tumour microenvironment.

Keywords:

• oxidative stress
• nitrosative stress
• redox biomarkers
• colorectal cancer

Acknowledgments

We are grateful to Martin Lenkiewicz, MSc, for his language assistance.

Abbreviations

CRC, colorectal cancer; FRAP, ferric reducing antioxidant power; MPO, myeloperoxidase; NO, nitrogen oxide; PC, protein carbonyls; IMA, ischemia modified albumin; AGE, advanced glycation end products; 8-OHdG, 8-hydroxydeoxyguanosine; GLOBOCAN, Global Cancer Observatory; adc, adenocarcinoma; muc adc, mucinous adenocarcinoma; CIN-high, chromosomal instability high; MSI-high, microsatellite instability high; ROS, reactive oxygen species; RNS, reactive nitrogen species; TB, tumour budding; PDC, poorly differentiated clusters; APDC, areas of poorly differentiated components; PBS, phosphate-buffered saline; BHT, butylated hydroxytoluene; TPTZ, 2,4,6-Tri(2-pyridyl)-s-triazine; 2,4-DNPH, 2,4-dinitrophenylhydrazine; HAS, human serum albumin; NBT, nitro blue tetrazolium; FFI, furyl-furanyl-imidazole; CML, carboxymethyl lysine; RCRC, right-side colorectal cancer; LCRC, left-side colorectal cancer; NOX, NADPH oxidase; COX, cyclooxygenase; LOX, lipoxygenase; NOS, nitric oxide synthase; TNF-α, tumor necrosis factor-α; NFκB, nuclear factor kappa-B; RAGE, receptors for advanced glycation end products; 3-NT, 3-nitrotyrosine; HOCL, hypochlorous acid.

Data Sharing Statement

All data generated or analysed during this study are included in this published article (and its supplementary information files).

Ethics Approval and Consent to Participate

The study was approved by the Bioethics Committee of the Medical University of Bialystok, Poland (permission number APK.002.99.2021). After a detailed explanation of the purpose of our research and the possible risk, all the qualified patients agreed in writing to participate in the experiment.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors declare that they have no competing interests.