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Journal of Cancer Research and Therapeutics, Vol. 7, No. 2, April-June, 2011, pp. 143-147 Original Article Quantitative evaluation of serum fucose in oral squamous cell carcinoma patients Rajkumar N Parwani1, Simran R Parwani2 1 Department of Oral and Maxillofacial Pathology, Modern Dental College, Indore, India Code Number: cr11034 PMID: 21768700 Abstract Background: Cancer, a disorder of cellular behavior is characterized by the alteration of serum glycoproteins, which are composed of different monosaccharides. One of the monosaccharides is l-fucose, a methyl pentose, which is the terminal sugar in most of the plasma glycoproteins. Elevated levels of protein-bound fucose have been reported in various disease states as well as in malignancies.Aim: To ascertain the role of serum fucose as a biomarker and to correlate with other studies for its effective clinical application. Materials and Method: The study was carried out on 67 subjects, including 14 healthy individuals and 53 oral squamous cell carcinoma cases. The serum fucose level estimation was done based on the method as adopted by Winzler using cysteine reagent. Statistical analysis included Chi-square test, Karl Pearson correlation test and Student's t test to evaluate the significance and variability of values between groups. Results: Serum fucose levels were independent of age and sex. However, there was significant increase in mean serum fucose level of oral squamous cell carcinoma patients compared with healthy controls. The results correlated well with other studies. Conclusion: Serum fucose can be used as an effective diagnostic biomarker in oral squamous cell carcinoma patients. Keywords: Biomarker for oral carcinoma, cancer, oral squamous cell carcinoma, serum fucose Introduction Cancer cells synthesize a wide variety of biochemical products that can be detected in body fluids. Of predominance and with great significance are serum glycoproteins. [1] Cell surface glycoconjugates are important in relation to cancer because many of the altered properties of cancer cells are expressed at the cell surface. Glycoconjugate molecules expressed in the plasma membrane of mammalian cells have also been reported to be associated with cell-to-cell adhesion, tumor progression and metastasis. [2] The carbohydrate of the glycoprotein is composed of relatively small number of different monosaccharides. One of them is l-fucose, which is usually a terminal sugar. It is one of the essential sugars that the body requires for optimum function of cell-to-cell communication. Apart from fucose being a prospective tumor marker, it is found to be a powerful immune modulator as it is distributed in macrophages, which are important for immune function. [3],[4] Physiologically, it is present in low concentrations in serum but is increased in cancer and other diseases. It has been documented that tumor cells modulate their surface by increasing fucosylation levels to escape recognition, which contributes to several abnormal characteristics of tumor cells, such as decreased adhesion and uncontrolled tumor growth. [5],[6] This mechanism is not specific for any anatomical group of malignancy and includes oral carcinoma. Several studies have suggested that monitoring serum/tissue fucose levels could be a promising approach for the early detection, diagnosis and prognosis of various cancer types, including oral carcinoma. [7],[8],[9],[10],[11] Clinically susceptible lesion can be analyzed with biomarker along with routine tests. Thus, the present study was undertaken with the following aims and objective:
Materials and Method The study was carried out in the Department of Oral and Maxillofacial Pathology, Modern Dental College and Research Center, Indore, after clearance from institutional ethical committee vide certificate no. MDCRC/IEC-09/07. The duration of the study extended for 9 months. Subjects were selected randomly in the age group of 23-75 years. A written consent was obtained from all the participants. The study sample consisted of 67 subjects, including 14 healthy individuals and 53 squamous cell carcinoma cases. To determine normal serum fucose levels, 14 healthy individuals, including 7 males and 7 females in the age range 23-67 years, were selected from subjects visiting the college for minor problems such as restoration, prophylactic oral care and escorts to other patients. The patient sample consisted of 53 cases with 36 males and 17 females in the age range 35-75 years with clinical signs of oral squamous cell carcinoma. Common exclusion criteria for both the groups, that is, controls and oral carcinoma patients was present or past history of any major illness such as liver disease, tuberculosis, diabetes and hypertension. Healthy controls were further excluded on the basis of tobacco consumption and alcoholism. Patients were selected on the basis of oral lesions prior to undergoing treatment of any form. At the onset, detailed case history of the patient was recorded and subsequently subjected to biopsy procedure for histopathological confirmation. The patient was asked to sit in chair in a comfortable position. Blood was drawn from the healthy subjects for the purpose of the study, whereas in patients it was drawn as part of routine hematological investigation prior to biopsy. Blood was collected by venipuncture and transferred to sterilized plain tubes and allowed to clot at room temperature. Then the clot was separated and serum centrifuged at 3000 rpm for 15 min to get a clear serum sample. The sample was stored at 4°C and assays were performed within 48 h. The serum fucose level estimation was done based on the method of Dische and Shettles [12] as adopted by Winzler. [13] L-fucose was assayed by dissolving ethanol-precipitated proteins of serum in alkali, addition of ice-cold sulfuric acid water mixture with subsequent immersion of test tubes in boiling water bath followed by addition of cysteine reagent. Standard l-fucose was procured from Sigma Company MO, USA. After 60-90 min at room temperature, the optical density readings were obtained from Bechman′s Spectrophotometer at 396 and 430 nm. Two optical densities were used to correct for the color produced by hexoses under these conditions. Serum l-fucose levels were calculated as mg/100 mL [Figure - 1]. Statistical analysis Statistical analysis was carried out using MSTAT-C software. Statistical analysis included mean values, standard deviation, 95% confidence interval, coefficient of variation, Chi-square test, Karl Pearson correlation test and Student′s t test. Chi-square test was used to evaluate the relationship between sex and serum fucose level. Karl Pearson correlation test was used to test the relationship between age and serum fucose level. Student′s t test was applied to evaluate the significance and variability of serum fucose between groups. Results The mean serum fucose level in 14 healthy controls in the present series was 5.323 ± 6767 mg%. Fifty-three patients showed a mean serum fucose level of 15.34 ± 0.86 mg% [Table - 1]. Correlation coefficient for age and serum fucose irrespective of sex and disease was 0.028 with P > 0.05 suggesting no relationship. Histopathologically, lesions included 2 cases of verrucous carcinoma, 38, 9 and 4 each of well, moderately and poorly differentiated squamous cell carcinoma respectively [Figure - 2]. There was no relationship between age, sex and serum fucose level. However, there was almost a threefold increase in mean serum fucose level from healthy controls to patients. Moreover, histopathological grading and serum fucose levels were independent of each other [Table - 2]. Discussion The search for unique molecular alteration associated with malignant transformation has been pursued for decades by a legion of biochemists and immunologists. It has also become well known that cancer cells synthesize a wide variety of biochemical products that can be detected in body fluids. This has led to the development of a wide variety of biochemical methods to detect neoplasms in cancer patients. [1] Numerous observations indicate that glycoproteins are often elevated above the normal levels in sera of cancer patients as well as in sera of animals bearing experimental tumors. [14],[15],[16] Measurement of protein-bound carbohydrates of glycoproteins has been used as an index to glycoprotein levels. A more recent trend has been to use the amount of given monosaccharide as a measure of glycoproteins. One of the monosaccharide is l-fucose, a methyl pentose, which is a terminal sugar in most of the plasma glycoproteins. [4] Fucose is found in many glycolipids and glycoproteins, including several families of blood group antigens. [17] Changes have been detected in the fucosylation pattern of these molecules in the tissue of cancer patients due to fucosyl transferase activity, which is especially high in the serum of patients suffering from highly malignant or metastatic tumors. [18],[19] Mean serum fucose level in 14 control individuals in the present series was 5.323 ± 677 mg% with levels ranging between 4.259 and 7.124 mg%. The level obtained in the present study was very much similar to that obtained by other workers, [20],[21],[22],[23] although slightly higher levels have been obtained by some other workers. [13],[24],[25],[26],[27],[28],[29],[30],[31] Oral squamous cell carcinoma patients showed mean serum fucose level of 15.34 ± 0.86 mg%. There was almost threefold rise in the levels from healthy controls to patients. The application of t test showed that the difference was highly significant statistically (P < 0.01). Similarly, higher serum fucose levels compared with those of the controls have been observed by various other workers. [24],[25],[26],[27],[30],[31],[32] It was observed that there was no relationship of serum fucose levels with age and sex. The findings of the present study were in accordance with those of Sharma and Sur, [20] Arya and Bhatnagar, [21] Solanki et al. [22] as well as with other workers. Two cases that were initially diagnosed as squamous cell carcinoma were histopathologically confirmed as verrucous carcinoma. These cases showed significant elevation in serum fucose levels as compared with healthy controls. Relationship of serum fucose level was ascertained with grades of squamous cell carcinoma patients. Although there was a slight difference in the mean of various histopathological grades, the difference was not significant statistically. This may be due to smaller sample size. Moreover, serum fucose level may be affected by other criteria such as secondary infection, size of the lesion, extent of the lesion and so on. The cause of the rise in serum fucose is difficult to explain, especially as the normal source and metabolic control of glycoprotein is not yet clearly understood. Various views have been expressed by different authors. The increased levels of fucose have been attributed to tissue destruction and tissue proliferation. Seibert et al. [33] suggested that elevation of serum fucose merely reflects the occurrence of tissue destruction and release of preformed fucose at the site. However, Shetlar et al. [14] suggested that tissue proliferation rather than repair is a more probable cause for increase in serum fucose. With respect to oral carcinoma, the possibilities exist that it may be a cancer product filtering in the blood or manifestation of generalized effect of oral carcinoma on the body metabolism. Rise in serum fucose level is not specific for cancers, as elevated serum fucose levels have also been reported in various pathological states such as cirrhosis liver and meningitis, [20] rickets and osteomalacia, [34] tuberculosis, [33],[35] cardiovascular disorders [36] as well as in depressive disorders. [29] Also, it has been observed that the serum fucose level is raised in different groups of malignancies such as breast cancer, [21],[22],[23] ovarian cancer, [9],[32],[37] colorectal adenocarcinomas, [6],[7] leukemias [5],[11] as well as brain tumors. [38] Thus, it becomes imperative in evaluating serum fucose levels in oral carcinoma to exclude other degenerative and proliferative diseases. Furthermore, size of the lesion and secondary inflammation could alter these levels. Altered levels of serum protein-bound fucose may be an indicator of both tumor burden and inflammatory response. Elevated levels have been observed in older age group of cancer patients, in cancer patients with metastasis and advancing stages than in subjects without metastasis. [30] Serum fucose is considered a better biochemical marker than sialic acid level in oral squamous cell carcinoma. [31],[39] Some studies have concluded that it is most effective of the essential sugars when it comes to slowing the growth of cancer cells. [3,4] Decrease in serum fucose levels after treatment has been observed by different workers, although it is emphasized that follow-up period has to be long enough. Conclusion There was a significant elevation in the mean serum fucose level in oral squamous cell carcinoma patients as compared with healthy controls. There was no relationship of serum fucose level with histopathological grades as well as with age and sex. The elevated level in oral cancer patients is nonpathognomonic. However, in conjunction with clinical diagnostic procedures, it can be used as an effective biochemical indicator. Moreover, it is an easy, noninvasive and cost-effective technique. References
Copyright 2011 - Journal of Cancer Research and Therapeutics The following images related to this document are available:Photo images[cr11034t1.jpg] [cr11034f1.jpg] [cr11034t2.jpg] [cr11034f2.jpg] |
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