Assessment of serum copper, iron and immune complexes in potentially malignant disorders and oral cancer

Abstract Potentially malignant disorders (PMDs) of oral cavity and oral cancer remain a cause of serious concern despite intensive research and development. Diet and immunity have been identified to play a crucial role as modifying factors in these diseases. Our study intended to explore this relationship by estimating and comparing the serum levels of copper, iron and circulating immune complexes (CICs) in patients diagnosed with PMDs and oral cancer and normal healthy individuals. In this study, 40 histopathologically diagnosed cases of PMDs and oral cancer were included along with 30 healthy controls and 5 ml of venous blood was drawn using venipuncture. Serum estimation of copper, iron and CIC then followed using the colorimetric and spectrophotometric methods. The data obtained was subjected to statistical analysis using one way ANOVA and Pearson's Product-Moment Correlation Test. The mean serum copper level was measured as 138.98 ± 10.13µg/100ml in the PMD group and 141.99 ± 21.44 µg/100ml in the oral cancer as compared to 105.5 + 18.81µ/100ml in the controls. The mean serum CIC levels was highest in the oral cancer (9.65 ± 0.16OD470) followed by the PMD group (0.18 + 0.21 OD470) and least in the control group (0.048 ± 0.02OD470). Whereas, the serum levels of iron showed a significant decrease in the PMD group (110.9 ± 10.54 µg/100ml) and the oral cancer group (114.29 ± 25.83 µg/100ml) as compared with the control group (136.85 ± 14.48 µg/100ml). There was no positive correlation obtained between the three groups with respect to the chosen parameters indicating that the variables were independent of each other. It can be thus be ascertained that trace elements like copper and iron as well as humoral responses (CICs) have a close relationship with PMDs and oral cancers.

Serum big endothelin-1 as a biomarker in oral squamous cell carcinoma patients: an analytical study

Detection of abnormally elevated levels of molecules in patients with oral cancer may be useful in early diagnosis. These markers can be included in current Histopathology grading and in TNM staging systems of Oral Squamous Cell Carcinoma (OSCC) to make it more efficient. Several pro-angiogenic molecules have been assessed for the same reason. Endothelin-1 (ET-1) is a vasoactive peptide associated with the development and spread of many solid tumors, including Squamous Cell Carcinoma (SCC), but its utility in OSCC has not been confirmed.Objective This study aims to evaluate the role of the serum big ET-1 as a biomarker of OSCC, by correlating it with the clinical staging and the histopathological grading.Material and Methods Serum levels of big ET-1 measured by the sandwich Enzyme-Linked Immunosorbent Assay (ELISA) in 40 OSCC cases were compared with the levels from the control group using independent t-test. Clinical stages and histopathological grades of OSCC cases were compared in relation to their mean levels of serum big ET-1, one using the Analysis of Variance (ANOVA) test and the other the independent t-test, respectively. The significance of the mean difference between the groups was evaluated by Tukey’s multiple comparison test. All statistical analyses were performed on GraphPad statistical software version 5.0.Results By comparing the mean of the big ET-1 concentrations of cases and controls, the independent t-test revealed significant higher big ET-1 concentration of OSCC cases when compared to controls (p<0.0001). Tukey’s multiple comparison test also revealed statistically significant difference among all OSCC stages in relation to the mean levels of serum big ET-1. However, the mean of the big ET-1 concentrations of cases of grade I and of grade II did not differ statistically (p=0.729).Conclusion Serum big ET-1 levels may be useful as a diagnostic tool in OSCC and as an adjunct to OSCC staging. However, its use as a prognostic marker warrants larger prospective studies.

ABO blood groups and oral premalignancies: A clinical study in selected Indian population.

Background: The ABO blood group antigens are present on the surface of red blood cells and various epithelial cells. As the majority of human cancers are derived from epithelial cells, changes in blood group antigens constitute an important aspect of human cancers. The aim of the study was to establish clinical usefulness of ABO blood group as a predisposing factor in early diagnosis and management of patients with oral precancerous lesions/conditions. Materials and Methods: The study sample consisted of 50 control and 50 oral precancer (25 leukoplakia and 25 Oral Submucous Fibrosis) confirmed by histopathologic examination. All samples were subjected to blood group testing and their prevalence was compared by Z-test using STATA version 8. Results: The "A" blood group was prevalent among the precancerous group. Significant differences on prevalences of blood groups were found (P < 0.05) between control versus leukoplakia and OSMF. Interestingly, 24% gutka chewers who had higher number of grades of dysplasia were falling in "A" blood group. Conclusion: Blood group type should be considered along with other risk factors to understand the individual patient's risk and further studies in larger samples with inclusion of Rh factor is needed to elucidate the relationship with ABO blood group types.

El sistema lewis hoy. nuevos enfoques / The lewis system today. new approaches

La expresión de antígenos (Ags) Lewis depende de alelos heredados en loci independientes, el gen Secretor (SE) que codifica la fucosiltransferasa 2 (FUT2) y el gen Lewis (LE) que codifica la fucosiltransferasa 3 (FUT3). El gen Se codifica una glicosiltransferasa que adiciona una fucosa en la cadena precursora de tipo 1 formando el Ag H en secreciones y fluidos. Como los azúcares inmunodominantes del Ag A y B pueden ser agregados a la cadena H de tipo 1, la FUT2 también controla la expresión de Ag A y B en las secreciones. El gen se es un alelo no funcional. El gen Le codifica una transferasa diferente que adiciona una fucosa en el 2do carbono en el precursor de tipo 1. El alelo le no es funcional. Las FUT2 y FUT3 interactúan para la formación de Ags Lewis en secreciones y fluídos. Los Ags Lewis en los eritrocitos no son en realidad parte integral de la membrana, están adsorbidos sobre la superficie en forma pasiva a partir del plasma. Están ampliamente distribuidos en tejidos humanos, eritrocitos, endotelio, riñón, tracto genitourinario, epitelio gastrointestinal y son receptores para algunos patógenos. Los anticuerpos (Acs) anti-Lewis en general no son clínicamente significativos, aunque se han publicado algunos casos de reacciones transfusionales hemolíticas, enfermedad hemolítica fetoneonatal y rechazo de transplante renal. Este trabajo es una revisión sobre los Ags del Sistema Lewis enfocada hacia sus diferentes funciones biológicas y su importancia en campos variados fuera del Banco de Sangre y la Inmunohematología tradicional.

The expression of Lewis blood group antigens depends on the alleles inherited at independent loci, FUT2 Secretor gene (SE) and FUT3 Lewis gene (LE). The Se and Le alleles encode separate fucosyltransferases that interact to form Lewis antigens in secretions and fluids. The Lewis antigens on red blood cells are not integral to the membrane but are passively adsorbed from the plasma. The allele Se encodes a transferase that adds fucose to type 1 precursor chains in secretions and fluids to form type 1 H antigen. Because A and B terminal sugars may be added to type 1 H chains, FUT2 also controls A and B expression in secretions. The FUT2 allele se gen is a nonfunctional allele. The FUT3 allele Le encodes a transferase that adds a fucose in other position in type 1 H precursor. The FUT3 allele le gen is a nonfunctional allele. The Le antigens are widely distributed in human tissues and fluids and are receptors for some pathogenic organisms. Lewis antibodies are rare and clinically no significant, although there are some reports of hemolytic transfusion reactions, hemolytic disease of the newborn and renal transplant rejection. This review focuses on different biological functions of Lewis antigens and their importance in some fields other than Blood Banks and traditional.

Relationship between ABO blood groups and oral cancer.

Background: Cancer is a unique disease characterized by abnormal growth of cells which have the ability to invade the adjacent tissues and sometimes even distant organs. Oral cancer has multifactorial etiology and is significantly associated with risk factors of the individual's lifestyle, particularly, chronic use of tobacco, spicy food, alcohol and smoking. Many studies have indicated that genetic factors also have an influence on the etiology of cancer. Aim: To evaluate if any of the ABO blood groups are associated with an increased risk for oral cancer. Materials and Methods: The present study was conducted at Kidwai Memorial Institute of Oncology (KMIO), Bangalore, after obtaining permission from the Director of the institute. The study sample comprised 235 oral cancer patients and 812 controls. For statistical analysis, Chi-square test and odds ratio were used to assess the relationship between ABO blood groups and oral cancer. Probability level was fixed at ≤0.05. Results: It was found that people with blood group A had 1.46 times higher risk of developing oral cancer compared to people of other blood groups. Conclusion: By employing a simple blood grouping test during community field programs, people with blood group A in the age group of 40-59 years having tobacco chewing habits can be apprised that they are more at risk to develop oral cancer than people with other blood groups.

Serum albumin in poorly differentiated, oral squamous cell carcinoma: a possible diagnostic marker of significance / Albumina sérica em carcinomas de células orais escamosas e pobremente diferenciadas: um possível marcador de diagnóstico de significância

Objectives: Serum albumin is considered to be the most potent and abundant extra-cellular anti-oxidant thatmight have a protective role in the ongoing process of transition of the various oral pre-cancerous lesions andconditions into frank malignant degenerations. The aim of this study was to check the reliability of serum albuminas one of the diagnostic anti-oxidant parameter. Materials and methods: The study consisted of seraanalysis of albumin in the age and sex matched normal healthy adults and patients with histologically proven,poorly differentiated oral squamous cell carcinoma. The results were analyzed using Student’s t-test andwere averaged as mean ± standard deviation. In above test, p-values less than 0.05 were taken to be statisticallysignificant. The normality of data was checked before the statistical analysis was performed. Results:The study revealed variations in sera levels of albumin to be statistically significant with the mean level ofsera albumin to be 4.956 ± 1.0579 in controls as against 3.6933 ± 1.2177 in patients with histologically proven,poorly differentiated, oral squamous cell carcinoma. Conclusions: The results of the study emphasize theneed for more studies with larger sample sizes to be conducted before a conclusive role could be drawn infavor of sera levels of albumin as diagnostic markers of significance in oral squamous cell carcinoma.

Objetivos: A albumina sérica é considerada o antioxidante extracelular mais poderoso e mais abundante, que pode exercer um papel protetor no processo de transição das várias lesões e circunstâncias pré-cancerígenasorais em degenerações malignas. O objetivo deste estudo foi verificar a confiabilidade da albumina sérica como um dos parâmetros de diagnóstico antioxidante. Materiais e métodos: A análise consistiu no estudo da albumina sérica em pacientes adultos saudáveis normais, separados por idade e sexo, e em pacientes com carcinoma oral de células escamosas pobremente diferenciadas com evidência histológica comprovada. Os resultados foram analisados usando o teste t de Student, e as médias foram calculadas com ± desvio-padrão. No teste citado, os valores de p menores que 0,05 foram considerados estatisticamente significantes. A normalidade dos dados foi verificada antes da realização da análise estatística. Resultados: O estudo revelou diferenças estatisticamente significantes nos níveis de albumina sérica com o nível médio de 4.956 ± 1.0579 nos controle sem contraste com 3.6933 ± 1.2177 nos pacientes com carcinoma oral de células escamosas pobremente diferenciadas.Conclusões: Os resultados do estudo enfatizaram a necessidade de mais estudos com tamanhos de amostra maiores antes que um papel conclusivo possa ser atribuído, em favor dos níveis de albumina sérica,como marcador diagnóstico para o carcinoma oral de células escamosas.

Evaluation of circulating immune complexes and serum immunoglobulins in oral cancer patients - A follow up study.

Background: High serum immunoglobulins and circulating immune complexes (IgG, IgM, IgA and CIC) values in patients with cancer have been used as tumor markers. Hence, the aim of the study was to estimate these immunological markers in pre- and post-treatment phases with a follow-up of 3-24 months and to understand the prognostic significance of the same in patients with oral cancer. Materials and Methods: The malignancy group consisted of 56 patients with different stages (AJCC TNM) of oral cancer and 20 healthy control group. Samples were selected at random and subjected for sequential analysis of serum biochemical markers (IgG, IgA, IgM and CIC-circulating immune complexes levels) in the pre- and post-treatment period. Statistical method employed was the paired t test. Results: We observed significant elevated levels of all the immunological markers ( P < 0.01) when compared with the control group. Sequential analysis of these markers revealed significant reduction in immunological markers in stage I and II patients. On the contrary, stage III and IV patients showed remarkably elevated levels of IgA and CIC one year after initial treatment. Conclusions : All these immunological markers are indicative of tumor burden and Serum levels of CIC and IgA might be employed as prognostic indicators in oral cancer.

Serum level of galectin-3 in patients with oral squamous cell carcinoma

Galectin-3, a member of the family of beta-galactoside-binding animal lectins, has been implicated in tumor invasion and metastasis. Using an ELISA kit, we assessed the circulating levels of galectin-3 in sera from 40 oral squamous cell carcinoma [OSCC] patients and from 43 healthy controls. Serum galectin-3 levels in OSCC patients were significantly higher [5.1 +/- 2.5 ng/ml] when compared with healthy controls [2.6 +/- 3.07 ng/ml, P<0.0005]. There was no apparent correlation between serum galectin-3 concentration and clinico-pathological features such as stage, tumor size, nodal status, distant metastasis and histological grade. This result suggests that, in addition to other tests, measurement of serum galectin-3 concentrations can be utilized as an adjuvant test for establishing a diagnosis of OSCC

Reverse transcriptase polymerase chain reaction study to evaluate dissemination of cancer cells into circulation after incision biopsy in oral squamous cell carcinoma.

BACKGROUND: Tissue manipulation by incisions, radiotherapy, and palpation may lead to dissemination of cancer cells into circulation. Circulating cancer cells in blood play a central role in metastatic process. Their numbers can be very small and for their detection,reverse transcriptase polymerase chain reaction (RT-PCR) has been successfully used in this study. MATERIALS AND METHODS: To examine whether cancer cell dissemination results from incision biopsy, we tried to detect oral squamous cell carcinoma (OSCC) cells in the peripheral blood sample before and after incision biopsy by CK19 RT-PCR. The study group consisted of 25 OSCC patients and the control group consisted of five patients with oral submucos fibrosis and five with leukoplakia. Five ml of blood collected before and twice (15 and 30 min) after incision were used for CK19 RT-PCR. RESULTS: Four (16%) of 25 cases of OSCC were positive for CK19 transcripts in their peripheral blood drained 15 min after incision. CK19 transcripts were not detected in the control group. CONCLUSION: Surgical invasion, in the form of incisional biopsy, causes dissemination of cancer cells into circulation, resulting in increased risk of metastasis.

Serum total glutathione-s-transferase levels in oral cancer.

We conducted a study wherein serum total glutathione-s-transferase levels were measured in patients (n = 27) with various stages of biopsy proven oral cancer (squamous cell carcinoma) and age and sex matched healthy human volunteers (n=10). In all patients with oral cancer, serum total glutathione-s-transferase was measured before the onset of treatment. There was a significant increase in serum total glutathione-s-transferase levels in patients with stage IV oral cancer as compared to stage II (P = 0.001) and stage III (P = 0.002) oral cancer. This shows that alterations in serum total Glutathione-s-transferase levels may have a role in cancer progression.

Expresión antigénica ABH y carácter secretor en pacientes con lesiones orales precancerosas y cancerosas / Secretor status and ABH antigens expression in patients with oral lesion

Los antígenos ABH, productos de la interacción de dos sistemas genéticos Hh y ABO, están sujetos a leyes de herencia y pueden estar localizados no sólo en los eritrocitos sino también en la mayoría de las células humanas. El objetivo del este trabajo fue investigar la relación entre el carácter secretor de pacientes con lesiones orales pre-malignas y malignas y la expresión antigénica ABH en cortes histológicos de dichas lesiones. Se trabajó con muestras incluídas en tacos de parafina de pacientes con lesiones orales. Los pacientes fueron clasificados en 2 grupos a) lesiones pre-malignas y malignas y b) lesiones benignas. Se investigaron los antígenos ABH por la técnica de inmunoadherencia específica modificada. Se utilizó la adherencia al tejido vascular como control positivo y al tejido adiposo como control negativo. Los resultados fueron semicuantificados desde adherencia fuertemente positiva a negativa. El carácter secretor fue determinado por la técnica de inhibición de la hemaglutinación. En 21 de las 34 muestras se observó una débil expresión antigénica en áreas atípicas, y deleción total en las áreas histológicamente afectadas por neoplasia. En 8 muestras hubo pérdida total de los antígenos ABH tanto en áreas normales como patológicas, estos pacientes presentaron un mayor grado de malignidad y metástasis que aquellos que conservaron la antigenicidad. Los pacientes con lesiones orales pre-malignas y malignas presentaron un incremento del carácter no secretor (52,3 por ciento) respecto de la población control (19,5 por ciento) y de aquellos pacientes con lesiones orales benignas (15.4 por ciento). Se observó una importante asociación entre pacientes no secretores y deleción de los antigenos ABH en muestras de lesiones orales. Además, hemos encontrado, en el grupo no secretor, una mayor malignidad de las lesiones orales como así también una mayor presentación de displasia epitelial. El estudio del carácter secretor en los pacientes con lesiones orales...

Lipid peroxidation & antioxidants status in patients with oral squamous cell carcinoma.

BACKGROUND & OBJECTIVE: Overproduction of lipid peroxidation byproducts and disturbances in antioxidant defense system have been implicated in the pathogenesis of several diseases including oral cancer. Though several studies have been done on the level of lipid peroxidation and antioxidants in oral cancer patients, there are no reports in patients with various clinical stages of oral squamous cell carcinoma. We carried out this study to assess the level of oxidative stress in oral cancer patients with various clinical stages. METHODS: Blood samples of 48 adult male oral cancer patients with various clinical stages of oral cancer (stage II to stage IV, 16 of each) and 16 age and sex matched healthy subjects were collected. Plasma and erythrocytes levels of thiobarbituric acid reactive substances (TBARS), vitamin E, reduced glutathione (GSH), and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were assayed using specific colorimetric methods. The statistical comparisons were performed by one way analysis of variance (ANOVA) followed by Student's t-test. RESULTS: Elevated lipid peroxidation and decline in non-enzymatic and enzymatic antioxidants status were noticed in oral cancer patients as compared to healthy subjects. The TBARS levels were gradually increased whereas antioxidants were gradually reduced from stage II to stage IV of oral cancer patients. INTERPRETATION & CONCLUSION: The altered lipid peroxidation in plasma and erythrocytes of oral cancer patients may be related to their compensatory changes in the antioxidants defense system.

Alterations in plasma lipid profile patterns in head and neck cancer and oral precancerous conditions.

BACKGROUND: The changes in lipid profile have long been associated with cancer because lipids play a key role in maintenance of cell integrity. AIMS: The present study evaluated alterations in plasma lipid profile in untreated head and neck cancer patients as well as patients with oral precancerous conditions (OPC) and its association with habit of tobacco consumption. MATERIAL AND METHODS: This hospital-based case control study included 184 head and neck cancer patients, 153 patients with OPC and 52 controls. Plasma lipids including: (i) Total cholesterol, (ii) LDL cholesterol (LDLC), (iii) HDL cholesterol (HDLC) (iv) VLDL cholesterol (VLDLC) and (v) triglycerides were analysed by spectrophotometric kits. STATISTICAL ANALYSIS USED: Student's t-test was performed to compare mean values of the parameters. RESULTS: A significant decrease in plasma total cholesterol and HDLC was observed in cancer patients (P=0.008 and P=0.000 respectively) as well as in patients with OPC (P=0.014 and P=0.000, respectively) as compared to the controls. The plasma VLDL and triglycerides levels were significantly lower in cancer patients as compared to the patients with OPC (P=0.04) and controls (P=0.059). The tobacco habituates showed lower plasma lipid levels than the non-habituates. Our data strengthen the evidence of an inverse relationship between plasma lipid levels and head and neck malignancies as well as OPC. CONCLUSION: The lower levels of plasma cholesterol and other lipid constituents in patients might be due to their increased utilization by neoplastic cells for new membrane biogenesis. The findings strongly warrant an in-depth study of alterations in plasma lipid profile in head neck cancer patients.

Study of serum fucose and serum sialic acid levels in oral squamous cell carcinomia.

Serum fucose and sialic acid levels were determined in 50 oral cancer patients and 25 healthy controls. A statistically significant increase was noted in the study group. The increase in serum fucose level correlated well with the clinical staging in the study group whereas sialic acid did not. These values were independent of age, sex and histopathological grading. The result suggest that the serum fucose level is a better biochemical tumor marker than sialic acid level. However its usefulness may be limited if used judiciously to assess the prognosis of the disease.

A case control study of selenium in cancer.

As selenium has been shown to be a risk modifier of cancer at several sites in a case-control approach, selenium nutriture was studied in oral and oesophageal cancer patients. The mean selenium level in oral cancer patients was 102.6 ng/ml and in oesophageal cancer 103.3 ng/ml, while in the matched controls it was 117.5 and 116.2 ng/ml respectively. Relative risk estimates were 3.16 (P < 0.02) and 16.0 (P < 0.01) in oral and oesophageal cancer patients respectively. The risk appears to increase with selenium levels lower than 115 ng/ml.