ABSTRACT
AIMS: The aim of this study was to simultaneously analyze estrogen quinone-derived adducts, including 17ß-estradiol-2,3-quinone (E2-2,3-Q) and 17ß-estradiol-3,4-quinone (E2-3,4-Q), in human albumin (Alb) and hemoglobin (Hb) derived from breast cancer patients with five-year postoperative treatment without recurrence in Taiwan and to evaluate the treatment-related effects on the production of these adducts. SETTINGS AND DESIGN: CohortMethods and Material: Blood samples derived from breast cancer 5-year survivors without recurrence were collected. Albumin and hemoglobin adducts of E2-3,4-Q and E2-2,3-Q were analyzed to evaluate the degree of disposition of estrogen to quinones and to compare these adduct levels with those in patients before treatment. STATISTICAL ANALYSIS: All data are expressed as mean ± standard deviation of three determinations. We used Student's t-test to examine subgroups. Data were transformed to the natural logarithm and tested for normal distribution for parametric analyses. Linear correlations were investigated between individual adduct levels by simple regression. Statistical analysis was performed using the SPSS Statistics 20.0. RESULTS: Result confirmed that logged levels of E2-2,3-Q-derived adducts correlated significantly with those of E2-3,4-Q-derived adducts (correlation coefficient r=.336-.624). Mean levels of E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb in 5-year survivors were reduced by 60-70% when compared to those in the breast cancer patients with less than one year of diagnosis/preoperative treatment (P<.001). CONCLUSIONS: Our findings add support to the theme that hormonal therapy including aromatase inhibitors and Tamoxifen may dramatically reduce burden of estrogen quinones. We hypothesize that combination of treatment-related effects and environmental factors may modulate estrogen homeostasis and diminish the production of estrogen quinones in breast cancer patients.
Subject(s)
Breast Neoplasms , Breast Neoplasms/drug therapy , Breast Neoplasms/metabolism , Estradiol/metabolism , Estrogens/metabolism , Estrogens/therapeutic use , Female , Humans , Quinones/metabolism , SurvivorsABSTRACT
Elevation of naphthoquinones and estrogen quinones, which are reactive metabolites of naphthalene and estrogen, is thought to be an important indicator of naphthalene- and estrogen-induced carcinogenesis. We compared background levels of naphthalene and estrogen quinone-derived adducts in serum albumin (Alb) from 143 women with breast cancer and 119 healthy controls. Cysteinyl adducts of naphthoquinones, including 1,2-naphthoquinone (1,2-NPQ) and 1,4-naphthoquinone (1,4-NPQ), and estrogen quinones, including estrogen-2,3-quinones (E2-2,3-Q) and estrogen-3,4-quinones (E2-3,4-Q), were characterized after adduct cleavage. Levels of estrogen quinones and naphthoquinones were positively correlated in healthy controls, but not in breast cancer patients (p < 0.05). Compared with controls, levels of 1,2-NPQ and E2-3,4-Q were elevated by two- to ten-fold in cancer patients (p < 0.001). To explore the correlation between estrogen- and naphthalene-derived quinone adducts and disease status, we performed linear discriminant analysis of the ratio of 1,2-NPQ-Alb to (1,2-NPQ-Alb plus 1,4-NPQ-Alb) versus the ratio of E2-3,4-Q-2-S-Alb to (E2-2,3-Q-4-S-Alb plus E2-3,4-Q-2-S-Alb) in patients and controls. These two groups were separable using albumin adducts of estrogen quinones and naphthoquinones, with 99.6% overall correct classification rate (overall accuracy). The findings of this study suggest that differences in the disposition of estrogen and naphthalene, and the subsequent elevation of cumulative E2-3,4-Q and 1,2-NPQ may serve as biomarkers of breast cancer risk.
Subject(s)
Breast Neoplasms/blood , Breast Neoplasms/diagnosis , Estrogens/blood , Naphthalenes/blood , Adult , Aged , Biomarkers, Tumor , Breast Neoplasms/etiology , Case-Control Studies , Disease Susceptibility , Female , Gas Chromatography-Mass Spectrometry , Humans , Middle Aged , Prognosis , Risk , Young AdultABSTRACT
Cumulative estrogen concentration is an important determinant of the risk of developing breast cancer. Estrogen carcinogenesis is attributed to the combination of receptor-driven mitogenesis and DNA damage induced by quinonoid metabolites of estrogen. The present study was focused on developing an improved breast cancer prediction model using estrogen quinone-protein adduct concentrations. Blood samples from 152 breast cancer patients and 71 healthy women were collected, and albumin (Alb) and hemoglobin (Hb) adducts of estrogen-3,4-quinone and estrogen-2,3-quinone were extracted and evaluated as potential biomarkers of breast cancer. A multilayer perceptron (MLP) was used as the predictor model and the resultant prediction of breast cancer was more accurate than other existing detection methods. A MLP using the logarithm of the concentrations of the estrogen quinone-derived adducts (four input nodes, 10 hidden nodes, and one output node) was used to predict breast cancer risk with accuracy close to 100% and area under curve (AUC) close to one. The AUC value of one showed that both data sets were separable. We conclude that Alb and Hb adducts of estrogen quinones are promising biomarkers for the early detection of breast cancer.
Subject(s)
Biomarkers, Tumor/blood , Breast Neoplasms/blood , Estrogens/blood , Hemoglobins/metabolism , Models, Biological , Quinones/blood , Serum Albumin, Human/metabolism , Adolescent , Adult , Aged , Female , Humans , Middle Aged , Predictive Value of TestsABSTRACT
BACKGROUND: Apurinic/apyrimidinic (abasic/AP) sites are among the most common endogenous DNA lesions. AP sites, if not repaired, could result in genomic instability as well as chromosome aberrations. Information regarding the direct assay of the number of abasic sites in human leukocytes and its association with risk of breast cancer has not been reported. METHODS: In this study, we investigated the association between certain risk factors for breast cancer and the background levels of AP sites in leukocytes derived from 148 Taiwanese women with breast cancer and 140 cancer-free controls. The risk factors studied include age, body mass index (BMI), and polymorphisms of apurinic/apyrimidinic endonuclease (APE1) [APE1 Asp148Glu(rs3136820)]. RESULTS: Mean levels of AP sites were estimated to be 23.3 and 50.3 per 106 nucleotides in controls and breast cancer patients, respectively (~twofold, p < 0.001). In subjects with age <50 or BMI < 27 (kg/m2), the levels of AP sites in breast cancer patients were ~2-3-fold greater than those of controls (p < 0.05). Additionally, results from the AP site 3'-cleavage assay indicated that the AP sites detected in both controls and patients were likely to be oxidant-mediated 5'-cleaved AP sites (~61-64 %). The number of AP sites in breast cancer patients was ~twofold greater in subjects with Asp/Glu + Glu/Glugenotypes than those with Asp/Asp genotype (p < 0.001). CONCLUSIONS: We confirmed that cumulative body burden of AP sites is a significant predictor of the risk of developing breast cancer and that genetic predisposition and environment factors may modulate the induction of oxidative DNA lesions in breast cancer patients.
Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/pathology , DNA-(Apurinic or Apyrimidinic Site) Lyase/genetics , Leukocytes/pathology , Polymorphism, Single Nucleotide , Adult , Case-Control Studies , DNA/metabolism , DNA Damage , Female , Gene Frequency , Genetic Predisposition to Disease , Humans , Middle Aged , Putrescine/metabolismABSTRACT
The objective of this research was to simultaneously analyze protein adducts of quinonoid metabolites of naphthalene and endogenous estrogen in serum albumin (Alb) derived from healthy pregnant women in Taiwan and to explore the correlations among them. The isomeric forms of cysteinyl adducts of naphthoquinones, including 1,2-naphthoquinone (1,2-NPQ) and 1,4-naphthoquinone (1,4-NPQ) as well as estrogen quinones, including estrogen-2,3-quinones (E2-2,3-Q) and estrogen-3,4-quinones (E2-3,4-Q), are characterized after adduct cleavage. Results showed that the median levels of cysteinyl adducts of 1,2-NPQ and 1,4-NPQ on serum albumin were 249-390 and 16.0-24.8 pmol g(-1), respectively. Logged levels of 1,2-NPQ-Alb were correlated with logged levels of 1,4-NPQ-Alb (correlation coefficient r = 0.551, P < 0.001). Cysteinyl adducts of E2-2,3-Q-1-S-Alb, E2-2,3-Q-4-S-Alb, and E2-3,4-Q-2-S-Alb were detected in all subjects with median levels at 275-435, 162-288, and 197-254 pmol g(-1), respectively. We also found a positive relationship between logged levels of E2-2,3-Q-4-S-Alb and those of E2-3,4-Q-2-S-Alb (r = 0.770, P < 0.001).We noticed that median levels of E2-2,3-Q-derived adducts (E2-2,3-Q-1-S-Alb plus E2-2,3-Q-4-S-Alb) in pregnant women were greater than those of E2-3,4-Q-2-S-Alb (â¼2-3-fold). Taken together, this evidence lends further support to the theme that cumulative concentration of E2-3,4-Q is a significant predictor of the risk of breast cancer. Furthermore, we noticed that levels of 1,2-NPQ-Alb are positively associated with levels of E2-3,4-Q-2-S-Alb (r = 0.522, P < 0.001) and those of E2-2,3-Q-4-S-Alb (r = 0.484, P < 0.001). Overall, this evidence suggests that environmental exposure to polycyclic aromatic hydrocarbons may modulate estrogen homeostasis and enhance the production of reactive quinone species of endogenous estrogen in humans.
Subject(s)
Breast Neoplasms/chemically induced , Estradiol/analogs & derivatives , Naphthalenes/metabolism , Polycyclic Aromatic Hydrocarbons/analysis , Quinones/metabolism , Serum Albumin/chemistry , Adult , Benzoquinones , Biomarkers/blood , Body Burden , Environmental Exposure , Estradiol/metabolism , Female , Humans , Naphthoquinones/metabolism , Pregnancy , TaiwanABSTRACT
The aim of this study was to establish a methodology to analyze estrogen quinone-derived adducts, including 17ß-estradiol-2,3-quinone (E2-2,3-Q) and 17ß-estradiol-3,4-quinone (E2-3,4-Q), in human hemoglobin (Hb). The methodology was then used to measure the levels of these adducts in Hb derived from female breast cancer patients (n=143) as well as controls (n=147) in Taiwan. Our result confirmed that both E2-2,3-Q- and E2-3,4-Q-derived adducts, including E2-2,3-Q-4-S-Hb and E2-3,4-Q-2-S-Hb, were detected in all breast cancer patients with median levels at 434 (215-1472) and 913 (559-2384) (pmol/g), respectively. Levels of E2-2,3-Q-4-S-Hb correlated significantly with those of E2-3,4-Q-2-S-Hb (r=0.622-0.628, p<0.001). By contrast, median levels of these same estrogen quinone-derived adducts in healthy controls were 71.8 (35.7-292) and 139 (69.1-453) (pmol/g). This translated to ~6-fold increase in mean values of E2-2,3-Q-4-S-Hb and E2-3,4-Q-2-S-Hb in breast cancer patients compared to those in the controls (p<0.001). Our findings add further support to the theme that cumulative body burden of estrogen quinones is an important indicator of breast cancer risk. We hypothesize that combination of genetic events and environmental factors may modulate estrogen homeostasis and enhance the production of estrogen quinones which lead to subsequent generation of pro-mutagenic DNA lesions in breast cancer patients.
Subject(s)
Biomarkers, Tumor/blood , Breast Neoplasms/blood , Estradiol/analogs & derivatives , Gas Chromatography-Mass Spectrometry/methods , Hemoglobins/metabolism , Adult , Aged , Estradiol/blood , Female , Humans , Limit of Detection , Linear Models , Middle Aged , Reproducibility of Results , Taiwan , Young AdultABSTRACT
Both 17ß-estradiol-2,3-quinone (E2-2,3-Q) and 17ß-estradiol-3,4-quinone (E2-3,4-Q) are reactive metabolites of estrogen. Elevation of E2-3,4-Q to E2-2,3-Q ratio is thought to be an important indicator of estrogen-induced carcinogenesis. Our current study compared the cumulative body burden of these estrogen quinones in serum samples taken from Taiwanese women with breast cancer (n=152) vs healthy controls (n=75) by using albumin (Alb) adducts as biomarkers. Results clearly demonstrated the presence of cysteinyl adducts of E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb in all study population at levels ranging from 61.7-1330 to 66.6-1,590 pmol/g, respectively. Correlation coefficient between E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb was 0.610 for controls and 0.767 for breast cancer patients (p<0.001). We also noticed that in premenopausal subjects with body mass index (BMI) less than 27, background levels of E2-3,4-Q-2-S-Alb was inversely proportional to BMI with about 25% increase in E2-3,4-Q-2-S-Alb per 5 kg/m(2) decrease in BMI (p<0.001). In addition, we confirmed that mean levels of E2-3,4-Q-2-S-Alb in breast cancer patients were â¼5-fold greater than in those of controls (p<0.001). Overall, this evidence suggests that disparity in estrogen disposition and the subsequent elevation of cumulative body burden of E2-3,4-Q may play a role in the development of breast cancer.
Subject(s)
Breast Neoplasms/blood , Estradiol/analogs & derivatives , Serum Albumin/metabolism , Adult , Biomarkers/blood , Body Burden , Body Mass Index , Case-Control Studies , Cell Transformation, Neoplastic/metabolism , Estradiol/blood , Female , Humans , Linear Models , Middle Aged , Protein Binding , Serum Albumin, Human , Taiwan , Up-RegulationABSTRACT
The role of fluoroquinolones (FQs) as empirical therapy for community-acquired pneumonia (CAP) remains controversial in countries with high tuberculosis (TB) endemicity owing to the possibility of delayed TB diagnosis and treatment and the emergence of FQ resistance in Mycobacterium tuberculosis. Although the rates of macrolide-resistant Streptococcus pneumoniae and amoxicillin/clavulanic acid-resistant Haemophilus influenzae have risen to alarming levels, the rates of respiratory FQ (RFQ) resistance amongst these isolates remain relatively low. It is reported that ca. 1-7% of CAP cases are re-diagnosed as pulmonary TB in Asian countries. A longer duration (≥ 7 days) of symptoms, a history of night sweats, lack of fever (> 38 °C), infection involving the upper lobe, presence of cavitary infiltrates, opacity in the lower lung without the presence of air, low total white blood cell count and the presence of lymphopenia are predictive of pulmonary TB. Amongst patients with CAP who reside in TB-endemic countries who are suspected of having TB, imaging studies as well as aggressive microbiological investigations need to be performed early on. Previous exposure to a FQ for >10 days in patients with TB is associated with the emergence of FQ-resistant M. tuberculosis isolates. However, rates of M. tuberculosis isolates with FQ resistance are significantly higher amongst multidrug-resistant M. tuberculosis isolates than amongst susceptible isolates. Consequently, in Taiwan and also in other countries with TB endemicity, a short-course (5-day) regimen of a RFQ is still recommended for empirical therapy for CAP patients if the patient is at low risk for TB.
Subject(s)
Community-Acquired Infections/drug therapy , Drug Resistance, Multiple, Bacterial , Fluoroquinolones/therapeutic use , Mycobacterium tuberculosis/pathogenicity , Tuberculosis, Pulmonary/drug therapy , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/therapeutic use , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Delayed Diagnosis , Endemic Diseases/prevention & control , Fluoroquinolones/adverse effects , Humans , Lung/drug effects , Lung/microbiology , Lung/pathology , Mycobacterium tuberculosis/drug effects , Taiwan/epidemiology , Time Factors , Tuberculosis, Pulmonary/epidemiology , Tuberculosis, Pulmonary/pathologyABSTRACT
Ornithine decarboxylase (ODC), the key enzyme of polyamine biosynthesis, has paradoxical roles in apoptosis. Our published papers show overexpression of ODC prevents the apoptosis induced by many cytotoxic drugs. Thapsigargin (TG) is an inhibitor of the sarcoplasmic/endoplasmic reticulum (ER) Ca(2+) ATPase (SERCA) pumps and causes ER stress-induced apoptosis. We used ODC overexpressing cell lines to examine whether overexpression of ODC inhibits TG-induced apoptosis. Our results indicated overexpression of ODC attenuated TG-induced apoptosis. Overexpression of ODC blocked procaspase-4 cleavage and phosphorylation of protein kinase-like ER-resident kinase (PERK), triggered by TG. It also attenuated the increase in CAAT/enhancer binding protein homologous protein (CHOP). Cells with overexpressed ODC had greater Bcl-2 expression. Overexpression of ODC preserved the expression of Bcl-2, inhibited the increase in Bak and stabilized mitochondrial membrane potential without the influences of TG. Cytochrome c release and down-stream caspase activation were blocked. That is, overexpression of ODC inhibits the mitochondria-mediated apoptotic pathway, induced by TG. Finally, overexpression of ODC maintains the protein and mRNA expression of SERCA. In conclusion, overexpression of ODC suppresses TG-induced apoptosis by blocking caspase-4 activation and PERK phosphorylation, attenuating CHOP expression and inhibiting the mitochondria-mediated apoptotic pathway.
Subject(s)
Apoptosis/drug effects , Apoptosis/physiology , Ornithine Decarboxylase , Thapsigargin/pharmacology , Animals , Caspase Inhibitors , Caspases/analysis , Caspases/metabolism , Cytochromes c/analysis , Cytochromes c/metabolism , Endoplasmic Reticulum/metabolism , Enzyme Inhibitors/pharmacology , Gene Expression Regulation , HL-60 Cells , Humans , Membrane Potential, Mitochondrial/drug effects , Membrane Potential, Mitochondrial/physiology , Mitochondria/metabolism , Ornithine Decarboxylase/genetics , Ornithine Decarboxylase/metabolism , Phosphorylation/drug effects , Phosphorylation/physiology , Polyamines/analysis , Polyamines/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism , Signal Transduction/drug effects , Signal Transduction/physiology , Transcription Factor CHOP/antagonists & inhibitors , Transcription Factor CHOP/metabolism , bcl-2 Homologous Antagonist-Killer Protein/antagonists & inhibitors , bcl-2 Homologous Antagonist-Killer Protein/metabolism , eIF-2 Kinase/metabolismABSTRACT
OBJECTIVE: A novel T-77C polymorphism in the promoter region of the DNA repair gene X-ray cross-complementing group 1 (XRCC1) may modulate its transcription to increase the risk of lung cancer. Here, we attempt to clarify: (i) whether the XRCC1 T-77C polymorphism was associated with lung cancer risk in Taiwanese and (ii) whether this polymorphism could act as a prognostic indicator to predict the clinical outcome of non-small-cell lung cancer (NSCLC) patients. METHODS: A total of 294 primary lung cancer patients and 288 potential controls were recruited into our study. Clinical data were collected. The genotypes of XRCC1 T-77C were identified by polymerase chain reaction. RESULTS: Our case-control study showed that the XRCC1 T-77C polymorphism was not associated with the risk of lung cancer in Taiwanese patients. To verify the impact of the XRCC1 T-77C polymorphism on the clinical outcome of NSCLC, survival analysis showed that patients with TT had a lower survival rate than those with the TC + CC genotype (33.1% versus 48.8%, P = 0.031). The Cox regression analysis further indicated that patients with the TT genotype had a 1.84-fold risk compared with those with the TC + CC genotype (95% CI, 1.16-2.86, P = 0.008). CONCLUSION: Our results suggest that XRCC1 T-77C variants (TC + CC) may act as a favorable prognostic indicator of resected NSCLC.
Subject(s)
Carcinoma, Non-Small-Cell Lung/genetics , DNA-Binding Proteins/genetics , Lung Neoplasms/genetics , Polymorphism, Genetic , Adenocarcinoma/genetics , Adenocarcinoma/mortality , Adult , Aged , Aged, 80 and over , Asian People , Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/mortality , Female , Humans , Lung Neoplasms/mortality , Male , Middle Aged , Polymerase Chain Reaction , Prognosis , Proportional Hazards Models , Regression Analysis , Taiwan , Transcription, Genetic , X-ray Repair Cross Complementing Protein 1ABSTRACT
BACKGROUND: Inconsistency in the prevalence of infection by human papillomavirus (HPV) in lung cancer patients was found between different countries with racial and geographic variations. Our previous reports have indicated that a high-risk HPV 16/18 DNA was frequently detected in Chinese lung cancer patients living in Taichung, Taiwan (Cheng et al. Cancer Res. 2001;61:2799-803). Thus, we conducted this study to verify whether there was a similar HPV 16/18 infection prevalence in lung cancer patients from Wuhan, China. METHODS: To reduce the false positive HPV detection, the paraffin sections of 73 lung tumors and 34 non-cancer controls from Wuhan, China were collected for detection of the presence of HPV 16/18 DNA by in situ hybridization (ISH). RESULTS: Our results showed that the rates of HPV 16 and/or 18 infections in patients with lung tumors were significantly higher than in 34 non-cancer control subjects (26.0 versus 2.8% for HPV 16, P = 0.030; 23.3 versus 5.7% for HPV 18, P = 0.031; 27.7 versus 5.9% for HPV 16 or 18, P = 0.003) with a similar infection frequency of HPV 16 and 18 types in lung tumors. This result indicated that HPV 16/18 infection may be associated with lung cancer development in Chinese patients from Wuhan, China. Further statistical analyses revealed that HPV 16 or 18 infection was not correlated with any clinico-pathological parameter studied, including age, gender, smoking status, tumor type, tumor stage and tumor grades. Interestingly, smoking and male patients had a higher prevalence of HPV 16, although not reaching a statistical significance, compared with non-smoking and female patients, respectively (33.3% for smokers versus 20.0% non-smokers; 33.3% for male versus 17.6% for female). As compared with the HPV 16/18 infection in Taiwan, Chinese patients with lung cancer from Wuhan had a different HPV 16/18 infection prevalence. CONCLUSION: Difference in HPV 16/18 infection in lung cancer patients from Wuhan, China and Taichung, Taiwan suggests that HPV 16/18 might play a different role in lung cancer development among Chinese living in different areas.