|Year : 2011 | Volume
| Issue : 2 | Page : 143-147
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
2 Department of Periodontics, Modern Dental College, Indore, India
|Date of Web Publication||12-Jul-2011|
Rajkumar N Parwani
Department of Oral and Maxillofacial Pathology, Modern Dental College, Indore
Source of Support: None, Conflict of Interest: None
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: T0 he 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
|How to cite this article:|
Parwani RN, Parwani SR. Quantitative evaluation of serum fucose in oral squamous cell carcinoma patients. J Can Res Ther 2011;7:143-7
| > 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.  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. 
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. , 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. , 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. ,,,, Clinically susceptible lesion can be analyzed with biomarker along with routine tests. Thus, the present study was undertaken with the following aims and objective:
- To evaluate and compare serum fucose levels in healthy controls and oral squamous cell carcinoma patients.
- To ascertain the role of serum fucose as an effective biomarker.
| > 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  as adopted by Winzler. 
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].
|Figure 1: Photograph showing test tubes with colored solutions of different groups after completion of test. (Std., standard; N, normal controls and M, malignant)|
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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.
|Table 1: Evaluation and comparison of mean serum fucose level of controls and oral squamous cell carcinoma patients|
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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. 
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. ,,
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.  Fucose is found in many glycolipids and glycoproteins, including several families of blood group antigens.  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. ,
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, ,,, although slightly higher levels have been obtained by some other workers. ,,,,,,,,
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. ,,,,,,
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,  Arya and Bhatnagar,  Solanki et al.  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.  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.  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,  rickets and osteomalacia,  tuberculosis, , cardiovascular disorders  as well as in depressive disorders.  Also, it has been observed that the serum fucose level is raised in different groups of malignancies such as breast cancer, ,, ovarian cancer, ,, colorectal adenocarcinomas, , leukemias , as well as brain tumors.  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. 
Serum fucose is considered a better biochemical marker than sialic acid level in oral squamous cell carcinoma. , 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|| |
|1.||Tormey DC, Davis TE, Walkers PT. Tumor Markers in Principles of Cancer treatment. New York: Academic Press; 1982. |
|2.||Kanagi R. Carbohydrate mediated cell adhesion involved in hematogenous metastasis of cancer. Glycoconj J 1997;14:577-84. |
|3.||Emil IM, Kitei M. Sugar that heal. The New Healing Science of Glyconutrients. New York: Balantine Publishing; 2001. p. 255. |
|4.||Elkins, Rita MH. Miracle Sugars: The Glyconutrient link to Better Health. Pleasant Grove, Utah, USA: Woodland Publishing; 2003. p. 220. |
|5.||MacDougall SL, Schwarting GA, Parkinson D, Sullivan AK. Increased fucosylation of glycolipids in a human leukaemia cell line (K562-Clone I) with decreased sensitivity to NK-mediated lysis. Immunology 1987;62:75-80. |
|6.||Youakim A, Herscovics A. Cell surface glycopeptides from human intestinal epithelial cell lines derived from normal colon and colon adenocarcinomas. Cancer Res 1985;45:5505-11. |
|7.||Fernandez-Rodriguez J, Paez de la Cadena M, Martinez-Zorzano VS, Rodriguez-Berrocal FJ. Fucose levels in sera and in tumours of colorectal adenocarcinoma patients. Cancer Lett 1997;121:147-53. |
|8.||Rao VR, Krishnamoorthy L, Kumaraswamy SV, Ramaswamy G. Circulating levels in serum of total sialic acid, lipid associated sialic acid, and fucose in precancerous lesion and cancer of the oral cavity. Cancer Detect Prev 1998;22:237-40. |
|9.||Wang JW, Ambros RA, Weber PB, Rusano TG. Fucosvltransferase and alpha-L-fucosidase activities and fucose levels in normal and malignant endometrial tissue. Cancer Res 1995;55:3654-8. |
|10.||Patel PS, Baxi BR, Adhvaryu SG, Balar DB. Evaluation of serum sialic acid, heat stable alkaline phosphatase and fucose as markers of breast carcinoma. Anticancer Res 1990;10:1071-4. |
|11.||Patel PS, Adhvaryu SG, Balar DB, Parikh BJ, Shah PM. Clinical application of serum levels of sialic acid, fucose and seromucoid fraction as tumour markers in human leukemias. Anticancer Res 1994;14:747-51. |
|12.||Dische Z, Shettles LB. A specific color reaction of methyl pentoses and a spectrophotometric micromethod for their determination. J Biol Chem 1948;175:595-604. |
|13.||Winzler RJ. Determination of serum glycoproteins In: Glick D, editor. Methods of biochemical analysis. New York: Interscience Publishers Inc; 1955; p. 279-311. |
|14.||Shetlar MR, Foster JV, Kelly KH, Shetlar CL, Bryan RS, Everett MR. The serum polysaccharide level in malignancy and other pathological states. Cancer Res 1949;9:515-9. |
|15.||Catchpole HR. Serum and tissue glycoproteins in mice bearing transplantable tumors. Proc Soc Exp Biol Med 1950;75:221-3. |
|16.||Apffel CA, Peters JH. Tumors and serum glycoproteins. The Symbodies. Prog Exp Tumor Res 1969;12:1-54. |
|17.||Vanhooren PT, Vandamme EJ. L-Fucose occurrence, physiological role, chemical, enzymatic and microbial synthesis. Chem Technol Biotechnol 1999;74:479-97. |
|18.||Oksana PY, Rima EP, Igor YA, Alexander SA. Fucose specific lectins in cancer research and diagnosis. Drug Des Rev 2005;2:349-59. |
|19.||Sen U, Guha S, Chowdhury JR. Serum fucosyl transferase activity and serum fucose levels as diagnostic tools in malignancy. Acta Med Okayama 1983;37:457-62. |
|20.||Sharma NC, Sur BK. Serum fucose and sialic acid levels in Indian children and adults under normal and pathological conditions. Indian J Med Res 1967;55:380-4. |
|21.||Arya DB, Bhatnagar KK. Evaluation of serum fucose level. Indian J Surg 1974;36:224-8. |
|22.||Solanki RL, Ramdev IN, Sachdev KN. Serum protein bound fucose in the diagnosis of breast malignancy. Indian J Med Res 1978;67:786-91. |
|23.||Kaswan HS, Kaushik SK. Serum fucose levels in the diagnosis of carcinoma breast. Indian J Surg 1982;44:741-3. |
|24.||Agarwal DP, Punia DP, Nawalkha PL, Khuteta KP. Effect of therapeutic irradiation on serum fucose levels in patients of oral cancer. Indian J Radiol 1980;34:255-9. |
|25.||Sen R, Sur R, Dasgupta R, Muzumdar GC. Serum pseudocholinesterase activity and protein bound fucose level in oral malignancy. Indian J Cancer 1987;24:242-9. |
|26.||Ghosh M, Raghavan RV, Nayak BR, Bailoor DN. Evaluation of serum fucose levels in patients with oral cancer. Oral Surg Oral Med Oral Pathol 1988;65:418-20. |
|27.||Ghosh M, Nayak BR. Serum sialic acid, fucose, sialic acid/fucose ratio as tumor marker in oral cancer. Ann Dent 1992;2:33-5. |
|28.||Macbeth RA, Bekesi JG. Plasma glycoprotein in various disease states including carcinoma. Cancer Res 1962;22:1170-4. |
|29.||Nandave M, Ojha SK, Kaur R. Changes in levels of serum glycoproteins in major depressive disorders. Indian J Clin Biochem 2005;20:154-7. |
|30.||Sawke NG, Sawke GK. Serum fucose level in malignant diseases. Indian J Cancer 2010;47:452-7. |
|31.||Shashikant MC, Rao BB. Study of serum fucose and serum sialic acid in oral squamous cell carcinoma. Indian J Dent Res 1994;5:119-24. |
|32.||Dutta TK, Sengupta U, Gupta BD. Clinical evaluation of serum protein bound fucose as a diagnostic and prognostic index in malignant tumors. Indian J Cancer 1976;13:262-6. |
|33.||Seibert FB, Seibert MV, Atno AJ, Campbell HW. Variation in protein and polysaccharide content of sera in the chronic diseases, tuberculosis, sarcoidosis and carcinoma. J Clin Invest 1947;26:90-102. |
|34.||Sharma NC, Sur BF. Serum fucose and sialic acid in rickets and osteomalacia. Clin Sci 1969;36:317-21. |
|35.||Mehta NG, Venkataraman A. Serum glycoprotein levels and diseases: An evaluation. Indian J Med Res 1975;63:970-7. |
|36.||Anand VK, Solanki RL, Ramdeo IN, Tandom SK. Serum protein bound fucose in cardiovascular disorders. Indian Pathol Bacteriol 1975;18:16-9. |
|37.||Aranganathan S, Senthil K, Nalin N. Case control study of glycoprotein status in ovarian carcinoma. Clin Biochem 2005;38:535-9. |
|38.||Manjula S, Monteiro F, Rao AA, Rao S, Annarwamy R, Rao A. Assessment of serum L-fucose in brain tumor cases. Ann Indian Acad Neurol 2010;13:33-5. |
|39.||Chitra S, Shyamala Devi CS. Effect of vitamin E on protein bound carbohydrate complexes in radiation treated oral squamous cell carcinoma patients. Indian J Clin Biochem 2008;23:92-4. |
[Figure 1], [Figure 2]
[Table 1], [Table 2]
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