Protein Thiols and Butryrylcholinestrase in Saliva of Oral Cancer Patients

Oral squamous cell carcinoma is one of the most common malignancies recognized. Biomarkers which can predict presence of cancer and its progression can help in better management of these disorders. Over production of lipid peroxidation byproducts and disturbances in antioxidant defense system have been implicated in the pathogenesis of several diseases including oral cancer. Studies have shown a correlation of butyrylcholinesterase (BChE), with tumourigenesis, cell proliferation and cell differentiation. Earlier we have observed a significant elevation in plasma BChE and protein thiols in oral cancer patients which correlated well with stages of cancer. As it was not clear whether the above markers will be altered in saliva of oral cancer patients this study was undertaken. Institutional Ethics Committee gave permission to carry out this study. Total of 55 subjects comprising healthy controls (n = 30) and biopsy proven oral cancer patients (n = 25) consented to participate in this study. Salivary samples from cases were taken before any definitive treatment. Protein thiols and BChE were estimated in salivary samples using validated assay methods. Oral cancer patients had a significant increase in pre-treatment salivary BChE levels (p ≤ 0.001) and a significant decrease (p ≤ 0.001) in salivary thiols as compared to respective values in controls. Salivary protein thiols and BChE may have a role in pathophysiology of oral cancer. Saliva can be used as a potential non-invasive screening tool in oral cancer patients.     

Effect of vitamin E on protein bound carbhohydrate complexes in radiation treated oral squamous cell carcinoma patients

Serum glycoproteins were evaluated in oral squamous cell carcinoma patients treated with radiotherapy and also the effect of vitamin E was studied. Cell surface glycoconjugates are important parameters in the detection of malignancy. Thus, the objective of the present study is to evaluate the efficacy of vitamin E on glycoproteins in oral cavity cancer patients treated with radiotherapy. The study includes 26 age and sex matched normal healthy individuals and 26 patients with squamous cell carcinoma of oral cavity. These patients were divided into two groups, one for radiotherapy alone (at a dosage of 6000 cGy in five fractions per week for a period of six weeks) and the other for radiotherapy plus vitamin E supplementation (at a dosage of 400 IU / day of vitamin E) for the entire period of radiotherapy. Levels of hexose, hexosamine, fucose and sialic acid were increased in oral squamous cell carcinoma patients and a significant decrease was observed in radiation treated patients when compared to control. The levels of glycoconjugates were significantly decreased in radiation treated patients supplemented with vitamin E. This measurement may be useful in assessing disease progression and identifying patients resistant to therapy and a possible role of vitamin E on reduction in glycoconjugate levels of radiation treated oral squamous cell carcinoma patients.     

Role of Oncofetal Protein CR-1 as a Potential Tumor Marker for Oral Squamous Cell Carcinoma

Cripto-1 (CR-1) is an oncofetal protein with its role as a key factor in early process of carcinoma has been evaluated in cases of various cancers. However, very few studies have reported its role in oral cancer, which is the sixth most common cancer around the world, particularly with high prevalence in developing countries. Oral squamous cell carcinoma (OSCC) is the most predominant (90%) of all the histological types of oral cancer. Late detection, associated with increased morbidity and mortality, is mainly attributed to non-availability of a suitable biomarker for the disease. In the present pilot study, we have evaluated the role of soluble CR-1, in serum as a potential tumor marker for OSCC. CR-1 was estimated using sandwich ELISA in serum samples of 50 biopsy proven OSCC patients (pre and post treatment) along with age and gender matched healthy controls. Immunohistochemistry was also done in corresponding tumor tissue sections to check the expression of CR-1. Pre-treatment CR-1 was found to be 2.25-fold higher in serum of OSCC patients as compared to control (p < 0.0001***), which was reduced to 1.6 folds post treatment (p = 0.0006***). CR-1 levels were comparatively higher in early stage of disease. Upon IHC 80% of the cases were found to be positive for CR-1. This study provides evidence that serum levels of CR-1 are elevated in patients of Oral Squamous Cell Carcinoma, which decrease post treatment. Also, the association of expression of protein with tumor progression predicts CR-1 as a molecule that can be further evaluated as a potential tumor maker in OSCC.     

Alteration of serum beta 2-microglobulin in oral carcinoma

Serum β₂-microglubin (β₂-m) levels were measured in oral carcinoma patients and compared with normal healthy controls. It was observed that there was a significant rise in serum β₂-microglobulin in oral carcinoma patients. Progressively higher values were obtained as the cancer advanced clinically. Therefore the estimation of serum β₂-microglobulin may be useful as one of a battery of tests in the assessment of oral carcinoma patients.     

宫颈鳞状上皮癌变过程中细胞周期调节通路p14^ARF-MDM2-p53作用的研究

目的 探讨细胞周期调节p14ARF-MDM2-p53通路在宫颈上皮癌变过程中的作用.方法 采用免疫组化SP法检测18例正常宫颈上皮、48例宫颈上皮内瘤变(CIN)和23例宫颈鳞状细胞癌组织p14ARF、MDM2及p53蛋白的表达.结果 p14ARF蛋白表达阳性率在正常宫颈上皮、CIN和宫颈鳞癌组分别为28%、42%和78%,宫颈鳞癌组表达较正常上皮和CIN组升高(P＜0.05);MDM2蛋白表达阳性率在正常宫颈上皮、CIN和宫颈鳞癌组分别为0%、23%和43%,CIN和宫颈鳞癌组较正常宫颈上皮表达升高(P＜0.05);p53蛋白表达阳性率在正常宫颈上皮、CIN和宫颈鳞癌组分别为6%、31%和39%,CIN及宫颈鳞癌组表达水平较正常宫颈上皮升高(P＜0.05);统计分析未发现三种蛋白的表达存在显著相关性.结论 p14ARF-MDM2-p53细胞周期调节通路与宫颈癌发生发展有关并有望用于CIN及宫颈癌的诊断.     

A report on clinical importance of serum glycoconjugates in oral cancer

For evolving a potential Biochemical index for early diagnosis of oral cancer, serum levels of total sialic acid (TSA), lipid bound sialic acid (LSA), mucoid proteins and hexoses (galactose and mannose) were determined in 30 patients with oral cancer. The results were compared with similar values obtained for 30 patients with oral precancerous conditions, 24 healthy controls and 15 normal individuals with a habit of tobacco chewing. All the biomarkers were significantly elevated in oral cancer patients compared to the controls, the normal chewers and precancer patients. The levels of glycoconjugates showed significant increase in patients with oral precancerous conditions compared to the controls. Only LSA and hexoses showed significantly elevated levels in oral precancer patients compared to the normal chewers. The levels of glycoconjugates were more or less comparable between normal chewers and the controls. The parameters evaluated in the oral cancer patients also showed a progressive rise with the clinical stage of the malignant disease. The levels of the four glycoconjugates were also higher in patients with metastasis compared to those with only primary disease. The present findings suggest that the glycoconjugates have clinical utility in diagnosis of the patients with cancer of the oral cavity as well as in staging the disease.     

A Review on Salivary Genomics and Proteomics Biomarkers in Oral Cancer

Oral cancer has emerged as an alarming public health problem with increasing incidence and mortality rates all over the world. Therefore, the implementation of newer screening and early detection approaches are of utmost importance which could reduce the morbidity and mortality associated with this disease. Sensitive and specific biomarkers for oral cancer are likely to be most effective for screening, diagnosis, staging and follow-up for this dreaded malignancy. Unlike other deep cancers, oral cancer is located in oral cavity. Hence, the direct contact between saliva and oral cancer lesion makes the measurement of tumor markers in saliva an attractive alternative to serum and tissue testing. The DNA, RNA and protein molecules derived from the living cancer cells can be conveniently obtained from saliva. Thus, salivary biomarkers, a non-invasive alternative to serum and tissue-based biomarkers may be an effective modality for early diagnosis, prognostication and monitoring post therapy status. In the current post-genomic era, various technologies provide opportunities for high-throughput approaches to genomics and proteomics; which have been used to evaluate altered expressions of gene and protein targets in saliva of oral cancer patients. The emerging field of salivary biomarkers has great potentials to prove its clinical significance to combat oral cancer. Hence, we have reviewed importance of several salivary genomics and proteomics biomarkers for oral cancer.     

Quick chip assay using locked nucleic acid modified epithelial cell adhesion molecule and nucleolin aptamers for the capture of circulating tumor cells

The role of circulating tumor cells (CTCs) in disease diagnosis, prognosis, monitoring of the therapeutic efficacy, and clinical decision making is immense and has attracted tremendous focus in the last decade. We designed and fabricated simple, flat channel microfluidic devices polydimethylsiloxane (PDMS based) functionalized with locked nucleic acid (LNA) modified aptamers (targeting epithelial cell adhesion molecule (EpCAM) and nucleolin expression) for quick and efficient capture of CTCs and cancer cells. With optimized flow rates (10 μl/min), it was revealed that the aptamer modified devices offered reusability for up to six times while retaining optimal capture efficiency (>90%) and specificity. High capture sensitivity (92%) and specificity (100%) was observed in whole blood samples spiked with Caco-2 cells (10–100 cells/ml). Analysis of blood samples obtained from 25 head and neck cancer patients on the EpCAM LNA aptamer functionalized chip revealed that an average count of 5 ± 3 CTCs/ml of blood were captured from 22/25 samples (88%). EpCAM intracellular domain (EpICD) immunohistochemistry on 9 oral squamous cell carcinomas showed the EpICD positivity in the tumor cells, confirming the EpCAM expression in CTCs from head and neck cancers. These microfluidic devices also maintained viability for in vitro culture and characterization. Use of LNA modified aptamers provided added benefits in terms of cost effectiveness due to increased reusability and sustainability of the devices. Our results present a robust, quick, and efficient CTC capture platform with the use of simple PDMS based devices that are easy to fabricate at low cost and have an immense potential in cancer diagnosis, prognosis, and therapeutic planning.     

Comparison of the Carcinogenic Potential of Smokeless Tobacco and Smoked Tobacco by Quantifying the Excretion of Nicotine Metabolite NNAL in Patients with Oral Leukoplakia

The case control study was conducted to quantify the amount of the carcinogenic tobacco specific nitrosamine in smokeless tobacco users and smokers in patients with oral leukoplakia and also to emphasize the role of tobacco-specific nitrosamine in the incidence of oral leukoplakia. The study was conducted on 30 patients. The urine samples were obtained from smokeless tobacco users with oral leukoplakia, smokers with oral leukoplakia and non-tobacco users (i.e. people who do not use tobacco in any form). The collected samples were processed by gas chromatography-mass spectrometry. A highly significant difference of NNAL[4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol] in smokeless tobacco users than tobacco smokers P = 0.0002 (Table 1) was obtained. This significant difference shows smokeless tobacco is more carcinogenic than smoked tobacco. This study confirmed that NNAL is a potent biomarker for calculating the risk of occurrence of carcinoma in smokeless tobacco users and smokers, and that smokeless tobacco is more harmful than smoked tobacco.     

Oxidatively Damaged DNA: A Possible Antigenic Stimulus for Cancer Autoantibodies

Reactive oxygen species (ROS) are cytotoxic at higher concentration resulting in cell death, mutations, chromosomal aberrations or carcinogenesis. In this study DNA was modified by singlet oxygen and superoxide anion radicals generated by illumination of riboflavin under 365 nm UV-light. The modified DNA induced high titre antibodies in experimental animals. In enzyme immunoassay, serum antibodies from cancer patients (n = 34) showed a higher recognition of the modified DNA, as compared to the native form. This was further confirmed by the gel-shift assay. Immune IgG were used as a probe to detect oxidative lesions in the DNA of cancer patients. DNA isolated from lymphocytes of cancer patients proved to be an appreciable inhibitor of the experimentally induced antibodies against the ROS-DNA. This indicates the presence of oxidative lesions in the DNA obtained from cancer patients. The results show that ROS induced oxidative damage to DNA in cancer patients generate neo-epitopes that are alien for the immune system, resulting in autoantibody formation.     

单克隆抗体MC3用于大肠癌和腺瘤的免疫组织化学研究及其预后意义

用单克隆抗体MC_3在福尔马林固定的48例大肠癌、31例癌变腺瘤、42例伴有不典型增生腺瘤、40例腺瘤、13例息肉以及48例正常肠粘膜组织切片上用免疫组化(PAP)法检测,结果MC_3相应抗原的表达阳性率分别为93.8％(45/48)、80.65％ (25/31)、47.62％ (20/42)、37.5％(15/40),其阳性率差异有非常显著性(P<0.001).48例正常肠粘膜均为阴性.MC_3在大肠癌组织中的反应程度与患者3年生存率有关,差异有显著意义(P<0.05).MC_3反应强度在高分化肠癌组与中～低分化肠癌组比较差异有显著意义(P<0.05).在绒毛状腺瘤、混合性腺瘤和管状腺瘤中MC_3相应抗原表达阳性率分别为71.4％(25/35)、60.0％(9/15)、41.3(26/63),差异有显著性(p<0.05).绒毛状腺瘤、混合性腺瘤和管状腺瘤其癌变率分别为45.7％(16/35)、13.3％(2/15)、20.6％(13/63),其癌变率比较差异有显著性(P<0.05).     

Clinical usefulness of alterations in sialic acid,sialyl transferase and sialoproteins in breast cancer

Sialic acid, the end moieties of the carbohydrate chains are biologically important and essential for functions of glycoconjugates and are reported to be altered in cancer patients. Two hundred and twenty five breast cancer (BC) patients, 100 patients with benign breast disease (BBD) and 100 healthy females (controls) were enrolled for the study. Eight hundred and twenty four follow-up samples of 225 breast carcinoma patients were also evaluated. The association of sialic acid forms, sialyltransferase and α-2–6 sialoproteins levels with presence and extent as well as prognosis of breast carcinoma was studied. Serum sialic acid forms and sialyltransferase revealed significantly elevated levels among untreated breast cancer patients as compared to the controls, patients with BBD as well as cancer patients in remission. Non-responders showed comparable levels of the markers with those found in breast cancer patients at the time of diagnosis. Higher levels of sialic acid forms at diagnosis were associated with poor prognosis. A positive correlation between serum levels of different forms of sialic acids and extent of malignant disease was observed. The changes in serum proteins with terminal α-2–6 sialic acid correlated well with alterations in the levels of sialic acid forms and sialyltransferase. Malignant tissues showed elevated levels of sialic acid and sialyltransferase as compared to surrounding normal tissues. The results suggested potential utility of these markers in evaluation of clinical outcome.     

Circulating peptide and steroid hormone levels in patients with breast cancer

Circulating levels of gonadotropins (FSH,LH), prolactin (PRL), estradiol and progesterone were measured by radioimmunoassay in the sera of 60 patients with histopathologically proven breast cancer. 30 patients belonged to the post and 30 to the premenopausal group. 30 healthy women, 15 each in the pre and post menopausal groups served as controls for the study. Prolactin was elevated in 33% and 26% of pre and post menopausal breast cancer patients respectively. Lower levels of estradiol and progesterone were observed during the luteal phase of the menstrual cycle in premenopausal breast cancer patients. Among postmenopausal patients with breast cancer, LH was elevated in 32%, FSH in 15% and estradiol in 30% of the patients studied.     

Electrotaxis of oral squamous cell carcinoma cells in a multiple-electric-field chip with uniform flow field

We report a new design of microfluidic chip (Multiple electric Field with Uniform Flow chip, MFUF chip) to create multiple electric field strengths (EFSs) while providing a uniform flow field simultaneously. MFUF chip was fabricated from poly-methyl methacrylates (PMMA) substrates by using CO₂ laser micromachining. A microfluidic network with interconnecting segments was utilized to de-couple the flow field and the electric field (EF). Using our special design, different EFSs were obtained in channel segments that had an identical cross-section and therefore a uniform flow field. Four electric fields with EFS ratio of 7.9:2.8:1:0 were obtained with flow velocity variation of only 7.8% CV (coefficient of variation). Possible biological effect of shear force can therefore be avoided. Cell behavior under three EFSs and the control condition, where there is no EF, was observed in a single experiment. We validated MFUF chip performance using lung adenocarcinoma cell lines and then used the chip to study the electrotaxis of HSC-3, an oral squamous cell carcinoma cell line. The MFUF chip has high throughput capability for studying the EF-induced cell behavior under various EFSs, including the control condition (EFS = 0).     

Polymorphisms of CYP1A1, GSTM1 and GSTT1 Loci as the Genetic Predispositions of Oral Cancers and Other Oral Pathologies: Tobacco and Alcohol as Risk Modifiers

Polycyclic aromatic hydrocarbons of tobacco require activation by phase I enzymes, such as cytochrome-P4501A1 (CYP1A1) to become an ultimate carcinogen, which are subjected to detoxification by phase II enzymes, especially glutathione S-transferases (GSTs). A study was designed to find whether genetic predisposition are risk modifiers of oral pathologies. The study included 102 cases with Oral Cancers (OCs), 68 cases with nonmalignant pathologies, 100 cases as control group. GSTM1 null genotype was associated with increased risk of OCs but not with benign pathologies. Deleted GSTT1 was associated with all pathologies. Both m1m2 and m2m2 polymorphisms of CYP1A1 were associated with oral pathologies.     

Diagnostic Role of Tumour Markers CEA, CA15-3, CA19-9 and CA125 in Lung Cancer

The aim of this study was to assess the diagnostic yield of the tumour markers carcinoembryonic antigen, carbohydrate antigen 15-3, carbohydrate antigen 19-9 and carbohydrate antigen 125, in serum and bronchoalveolar lavage fluid in a group of patients with bronchogenic carcinoma. Serum and bronchoalveolar lavage fluid samples were collected in a group of 90 patients with benign or malignant pulmonary diseases. After appropriate processing, tumour markers were determined by enzyme immunoassay. The diagnostic yields (sensitivity, specificity and predictive values) in each environment (serum and bronchoalveolar lavage fluid) were obtained by using "Receivers operating characteristic" curve. Determined individually, carcinoembryonic antigen, carbohydrate antigen 19-9 and carbohydrate antigen 125, showed the greatest diagnostic accuracy in bronchoalveolar lavage fluid. Carbohydrate antigen 15-3 did so in serum. Carcinoembryonic antigen was the most relevant marker in bronchoalveolar lavage fluid. For the factors evaluated in this study, determination of carcinoembryonic antigen, carbohydrate antigen 19-9 and carbohydrate antigen 125 in bronchoalveolar lavage fluid were clinically more useful markers in comparison with serum, although the latter may also be helpful in certain situations. Although there is no specific tumour marker for lung cancer, the combination of several can be used to diagnose most patients with lung cancer and also to rule out false positive and negative cases.     

氟他胺加睾丸切除治疗晚期前列腺癌22例报告

为探讨提高晚期前列腺癌的治疗效果，回顾性研究22例晚期前列腺癌的临床资料，运用氟他胺加睾丸切除治疗前列腺癌，其中C2期8例，D1期9例，D2期5例。结果：显效17例，有效3例，死亡2例，总有效率达90．9％。结果表明氟他胺是一个单纯抗雄激素的非类固醇化合物，对前列腺组织靶细胞膜上的雄激素受体有特异性阻断作用，能抑制雄激素依赖性肿瘤细胞生长，切除睾丸能降低雄激素水平，两者产生协同作用，提高疗效。     

Evaluation of serum glycoproteins in oral carcinoma

The present study was conducted on thirty untreated oral cancer patients proved by clinical and histopathological evidence and thirty healthy control subjects. The levels of glycoprotein-associated carbohydrates such as hexose, hexosamine, fucose and sialic acid were found to be elevated significantly as compared to control subjects. There was a progressive rise in these markers as the stages of oral cancer advanced.     

Sex hormone binding globulin in breast cancer

The present study was undertaken to determine the significance of sex hormone binding globulin, the major and specific binding protein for testosterone and estradiol, in breast cancer. Among breast cancer patients, lower serum levels of Sex hormone binding globulin and higher levels of testosterone were observed. Sex hormone binding globulin showed an inverse relationship with testosterone and total cholesterol, and a direct relation with HDL-cholesterol. By the western blot analyses, Sex hormone binding globulin was detected in all biological samples that we examined. In the breast tumor tissue sections, immuno-staining for Sex hormone binding globulin was confined in cell cytoplasm and 29% cases were positive, which showed no association with the investigated prognostic markers of breast cancer such as ER and HER-2/neu over-expression. In this study, decreased circulating levels of Sex hormone binding globulin in breast cancer patients possibly indicate higher bioavailable estrogens.