Clinical Study Relation Between Gastric Cancer and Protein Oxidation, DNA Damage, and Lipid Peroxidation
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Hindawi Publishing Corporation Oxidative Medicine and Cellular Longevity Volume 2013, Article ID 543760, 6 pages http://dx.doi.org/10.1155/2013/543760 Clinical Study Relation between Gastric Cancer and Protein Oxidation, DNA Damage, and Lipid Peroxidation Yongsheng Ma,1 Lin Zhang,1 Shengzhong Rong,2 Hongyan Qu,3 Yannan Zhang,4 Dong Chang,1 Hongzhi Pan,4 and Wenbo Wang1 1 The First Affiliated Hospital of Harbin Medical University, No. 199 Dongdazhi Street, Nangang District, Harbin, Heilongjiang 150001, China 2 Public Health School, Mudanjiang Medical College, No. 3 Tongxiang Street, Aimin District, Mudanjiang, Heilongjiang 157011, China 3 The Third Affiliated Hospital of Harbin Medical University, No. 150 Haping Road, Nangang District, Harbin, Heilongjiang 150081, China 4 Public Health School, Harbin Medical University, No. 157 Baojian Road, Nangang District, Harbin, Heilongjiang 150081, China Correspondence should be addressed to Hongzhi Pan; [email protected] and Wenbo Wang; [email protected] Received 28 August 2013; Revised 29 October 2013; Accepted 1 December 2013 Academic Editor: Neelam Khaper Copyright © 2013 Yongsheng Ma et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Objects. Theaimofthisstudyistoevaluateproteinoxidation,DNAdamage,andlipidperoxidationinpatientswithgastriccancer and to investigate the relationship between oxidative stress and gastric cancer. Methods. We investigated changes in serum protein carbonyl (PC), advanced oxidation protein products (AOPP), and 3-nitrotyrosine (3-NT) levels, as indicators of protein oxidation, serum 8-hydroxydeoxyguanosine (8-OHdG), as a biomarker of DNA damage, and malondialdehyde (MDA), conjugated diene (CD), 4-hydroxynonenal (4-HNE), and 8-ISO-prostaglandin F2 (8-PGF) in serum, as lipid peroxidation markers in gastric cancer (GC) patients and healthy control. Results. Compared with control, a statistically significant higher values of 8-OHdG, PC, AOPP, and3-NTwereobservedintheGCpatients( < 0.05). The products of lipid peroxidation, MDA, CD, 4-HNE, and 8-PGF, were significantly lower in the GC patients compared to those of control ( < 0.05). In addition, the products of oxidative stress were similar between the Helicobacter pylori positive and the negative subgroups of GC patients. Conclusions. GC patients were characterized by increased protein oxidation and DNA damage, and decreased lipid peroxidation. Assessment of oxidative stress and augmentation of the antioxidant defense system may be important for the treatment and prevention of gastric carcinogenesis. 1. Introduction Normal level of ROS has physiological functions, whereas it is toxic to the cells at high level. Overproduction of ROS Gastric cancer (GC) is one of the most frequent diseases througheitherendogenousorexogenousinsultsisharmful in human population. It is the fourth frequent cancer and to living organisms and is termed oxidative stress [3]. the second most common cause of deaths from cancer in Oxidative stress can damage cellular macromolecules, the world, accounting for a large proportion of cancer cases leading to DNA and protein modification and lipid perox- in China in recent years [1]. The pathogenesis of GC is not idation [4]. Oxidative stress produced by free radicals has understood completely. Nutritional, microbial, and genetic been associated with the development of several diseases factors acting in a multistep and multifactorial process have such as cardiovascular, cancer, and chronic inflammation3 [ – been proposed [2], where oxidative stress was involved in the 7]. It is also known that ROS could be excessively formed possible mechanisms of GC development. in chronic diseases of the gastrointestinal tract and toxic to Reactive oxygen species (ROS) are the normal product of normal cells, which might contribute to the increased risk of a variety of essential biochemical reactions and the genera- cancer [6, 7], but the precise mechanisms are still remaining tion of ROS is an unavoidable consequence of aerobic life. to be elucidated. 2 Oxidative Medicine and Cellular Longevity The aim of this study is to investigate the relationship 3 months. All samples from each patient were run in the same between oxidative stress and GC. We used spectrophotomet- assay. ric assay and enzyme-linked immunosorbent assay (ELISA) to detect the levels of lipid peroxidation products such 2.3. Laboratory Analysis. AOPP were quantified as described as malondialdehyde (MDA), conjugated dienes (CD), 4- by Witko-Sarsat et al. [8]. PC concentrations in plasma hydroxynonenal (4-HNE), and 8-Iso-prostaglandin F2 (8- were measured with spectrophotometric assay described by PGF) and measured the concentration of protein damage Reznick and Packer [9]. CD and MDA were measured by products including advanced oxidation protein products spectrophotometric assay [4]. Serum 8-OHdG was mea- (AOPP), protein carbonyl (PC), and 3-nitrotyrosine (3-NT) sured with ELISA method following the maker’s instructions in patients with gastric cancer and healthy control subjects. (Highly Sensitive 8-OHdG Check, Japan Institute for the In addition, we determined 8-hydroxydeoxyguanosine (8- Control of Aging, Fukuroi, Shizuoka). 3-NT (Hycult Biotech- OHdG) content, the DNA damage produced in GC patients, nology B.V.), 4-HNE, and 8-PGF (Cell Biolabs, Inc.) were andhealthycontrol,too.Inthepresentstudy,weprovide measured with ELISA following the maker’s instructions. evidence for the first time indicating the relationship between Briefly, 50–100 L of serum was added into each well of the ∘ proteincarbonylandtheriskofgastriccancer. microtiter plate and incubated for 2h at 37 C. Secondary ∘ antibodywasaddedtotheplateandincubatedat37Cfor 1 h, the unbound enzyme-labeled secondary antibody was 2. Material and Methods removed, and then the antibodies binding with the plate 2.1. Subjects. This is an age-and sex-matched case-control were identified using a substrate that contained 3,3 ,5,5 - study.ThirtypatientswithnewlydiagnosedGCarerecruited tetramethylbenzidine. Absorbance was measured using a for the experiment group, among whom 17 patients were spectrophotometric plate reader (BIO-RAD) at wavelength of + − positive for Helicobacter pylori (Hp ) and 13 negative (HP ). 450 nm. Quantification of the 8-OHdG, 3-NT, 8-PGF, and 4- Diagnosis of gastric cancer was confirmed by histopathologi- HNE were done by comparing the optical densities of each cal examination and histopathology. Tumors were classified sample to that of an internal standard known at various according to their predominant histological pattern into concentrations. intestinal (=13), diffuse (=6), and adenocarcinoma (=11). The pathologic stage (TNM) was determined 2.4. Other Biochemical Parameters. Blood glucose, triglyc- according to the newly revised classification of the American erides, and cholesterol were determined using routine clinical Joint Committee on Cancer (AJCC) and the International chemical assays. Union Against Cancer (UICC). Seven patients had AJCC Stage II tumors, and ten patients had AJCC Stage III tumors. 2.5. Statistical Analysis. Data are presented as mean ± SD. The remaining thirteen patients had UICC Stage III or All experimental data in this study were statistically analyzed IV gastric adenocarcinoma. Urease test and histopathologic with SAS 9.13. The statistical significance was evaluated using examination were carried out to determine the infection of unpaired Student’s t-test. Results were considered significant Helicobacter pylori (HP). If the urease test and histology when < 0.05. were positive, the diagnosis of HP infection was confirmed. − If both tests were negative, the patient was considered Hp . 3. Results Exclusion criteria included subjects who had been taking antioxidant supplements and regularly used acetylsalicylic 3.1. Baseline Characteristics. The clinical characteristics of the acid and other nonsteroidal anti-inflammatory drugs. Other GC patients and control subjects are shown in Table 1.Age exclusion criteria included diabetes mellitus, hypertension, and gender of the patients were not significantly different arteriosclerosis, severe liver disease, cataract, renal failure, from those of the controls. Initial clinical laboratory test and dialysis. All patients were recruited from the Third values, such as glucose, cholesterol, triglycerides, and blood Affiliated Hospital of the Harbin Medical University. Both pressure, were also not significantly different between the two groupsbelongedtothesameethnicgroup. groups. What is more, all the test values were in normal range. Thirty healthy, age-matched subjects, who came to the same hospital for an annual checkup, were also included as 3.2. Results of Protein Oxidative Damage and DNA Damage controls. The control subjects were in good health, as deter- Products in Serum. As shown in Table 2,theconcentration mined by a medical history questionnaire, physical examina- of AOPP in GC patients was increased significantly than that tion, and normal results of clinical laboratory tests. None of of control (25.75 ± 5.75 versus 22.29 ± 5.27; < 0.05). Serum the control subjects were taking any medication. All subjects 3-NT also significantly increased in GC patients compared gave written informed consent to participate in this study. with normal subjects (117.75 ± 37.12 versus 92.85 ± 14.47, < 0.01); so was serum PC (2.16 ± 0.68 versus