|Year : 2021 | Volume
| Issue : 1 | Page : 40-44
Role of sialic acid, protein, and sugar parameters of saliva and serum as a diagnostic biomarker tool among oral squamous cell carcinoma patients at rural Kanpur: An evaluative case—control assessment
Kriti Garg1, Rohan Sachdev2, Samiksha Shwetam1, Praveen Singh1, Akash Srivastava1
1 Department of Oral Medicine and Radiology, Rama Dental College, Kanpur, Uttar Pradesh, India
2 Department of Public Health, UWA School of Population and Global Health, University of Western Australia, Nedlands, Australia
|Date of Submission||06-May-2021|
|Date of Acceptance||01-Jun-2021|
|Date of Web Publication||12-Jul-2021|
Dr. Kriti Garg
117/K-68 Sarvodaya Nagar, Kanpur, Uttar Pradesh
Source of Support: None, Conflict of Interest: None
Context: It is essential to identify cancer at an early stage to reduce the disease's morbidity and mortality. Noninvasive methods such as serum and saliva analysis, in comparison to existing biopsy, can provide a cost-effective approach for screening a large population. Aims: The aim of this study was to evaluate sialic acid (SA), protein, and sugar in serum and saliva of oral squamous cell carcinoma (OSCC) patients with controls to assess their role as a diagnostic marker. Settings and Design: Unstimulated whole saliva and serum were collected. Subjects and Methods: Unstimulated whole saliva and serum were collected. From sixty controls and 78 squamous cell carcinoma patients. Serum and salivary total sialic acid, total protein, and total sugar estimation were carried out. Statistical Analysis Used: The results were tabulated and analyzed statistically using the t-square test, Pearson correlation with SPSS 21 version statistical software. Results: A significant rise in the salivary and serum SA as well serum protein was observed in OSCC study patients as compared to controls. Salivary protein, serum, and salivary sugar also show significant results. Furthermore, serum and salivary SA levels were found to be significantly increased with histopathological grading. Conclusions: The present study showed a significant and gradual increase in serum and salivary SA, protein, and sugar from control to oral cancer. From this study, we can suggest that SA can be used as a reliable biomarker in saliva for screening and early detection of oral cancer.
Keywords: Oral cancer, protein, saliva, serum, sialic acid, sugar
|How to cite this article:|
Garg K, Sachdev R, Shwetam S, Singh P, Srivastava A. Role of sialic acid, protein, and sugar parameters of saliva and serum as a diagnostic biomarker tool among oral squamous cell carcinoma patients at rural Kanpur: An evaluative case—control assessment. Indian J Oral Health Res 2021;7:40-4
|How to cite this URL:|
Garg K, Sachdev R, Shwetam S, Singh P, Srivastava A. Role of sialic acid, protein, and sugar parameters of saliva and serum as a diagnostic biomarker tool among oral squamous cell carcinoma patients at rural Kanpur: An evaluative case—control assessment. Indian J Oral Health Res [serial online] 2021 [cited 2022 May 19];7:40-4. Available from: https://www.ijohr.org/text.asp?2021/7/1/40/321115
| Introduction|| |
In India, 30%–40% of all malignant tumors responsible for the occurrence of head-and-neck cancer. Head-and-neck squamous cell carcinoma, with an estimated 1.5 million new cases diagnosed annually, ranks 6th globally for cancer-related mortality. The most problematic question is detecting and evaluating emerging oral cavity preliminary cancer. With the probability of a high recurrence rate and secondary metastasis, the clinical decision for the course of treatment and therapeutic approaches becomes more essential. To be scientifically valid, any biochemical agent must be associated with cancer cells and present in sufficient quantities in tissue or fluid where its concentration can be correlated to the underlying cancer progression. Its also worth mentioning that the biochemical alteration is a result of cancer, not something that occurred on its own or before the emergence of cancer symptoms., These biochemical substances are termed as tumor markers which can be defined as “A substance that is measured objectively and evaluated as an indicator of normal biologic processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention.”,
Sialic acid (SA) is a protein-bound monosaccharide which occurs in combination with other monosaccharides such as galactose, mannose, glucosamine, and fucose. It is a constituent of many proteins in saliva and is mainly present in salivary mucin. Its level is studied for the evaluation of synthesis and secretion of glycoprotein. It has been known to increase in patients with various premalignant and malignancies. Unnatural glycosylations are a universal feature of cancer, and the level of these glycoconjugates increases as cancer progresses. Several studies have shown that variations in serum SA levels in cancer patients correlate with a substantial reduction in the cancer cell, recurrence, and metastases, and have been recognized as a useful cancer marker in the clinical status of patients with carcinoma. Total sugar and total protein (TP) are standard tests performed in patients suspected of having a pathology, and any substantial changes to them, if found, can support the early detection of malignancy. The majority of recent research has focused on serum SA, but a few studies have used saliva as a diagnostic fluid for oral cancer. The effects of biochemical saliva analysis have been studied less in routine diagnostic settings. Saliva can be very beneficial because it can be obtained easily and noninvasively. Therefore, this study was undertaken to estimate salivary levels of SA, protein (P), sugar (S) in oral squamous cell carcinoma (OSCC) patients and compare them with serum levels to aid in assessing their role as diagnostic tumor marker for early detection of oral cancer and monitoring the progression of the disease at rural Kanpur.
| Subjects and Methods|| |
The present retrospective case—control study was carried out at dental college, in rural Kanpur, India. Informed consent was obtained from all the study patients and ethical approval was obtained from the ethical committee of the institute. It comprised a total of 138 patients, which included 78 clinically and histopathologically diagnosed cases of OSCC and sixty healthy age and gender-matched controls. The OSCC patients were categorized clinically according to the American joint committee on cancer tumor, lymph nodes, metastasis staging system, and histopathologically into well-differentiated, moderately differentiated, and poorly differentiated squamous cell carcinoma (PDSCC) according to Border's criteria, [Table 1].
|Table 1: Demographic details of oral squamous cell carcinoma study patients (n=78)|
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Serum and whole unstimulated saliva samples were collected from all the patients to analyze SA, protein, and sugar levels. To ensure minimal diurnal variation in composition, saliva was collected between 10 a. m. and 12 p. m., 2 h after the patient's usual breakfast time. Patients were told to rinse their mouths vigorously with clean water and to void their mouths of saliva before saliva collection The patients were asked to spit out their saliva into sterile plastic containers after it had accumulated on the floor of their mouth. The patients were told not to spit forcibly to prevent any possible blood contamination from the inflamed gingival tissue or ulcerated lesion. The plastic container was placed in an ice carrier box and transferred to the laboratory for biochemical analysis after saliva (2 ml) was collected. Venous sample was collected from the right-hand cubital vein under aseptic precautions. Both the saliva and blood samples were centrifuged at 3000 rpm for 15 min. The supernatant was taken for the estimation of SA, proteins, and sugar levels. Serum and salivary total SA level were determined by the method as described by Skoza and Mohos. Salivary proteins were estimated by the method of Lowry et al. Bovine serum albumin standards ranging in concentration from 0.05 to 0.5 mg/ml were done simultaneously with the test samples. Sugar content was estimated based on phenol-sulfuric acid reaction of sugars. Glucose standards ranging in concentration from 0.01 to 0.08 mg/ml, was done.
All the biochemical parameter values gathered were further tabulated and statistically analyzed using SPSS 21 version software (IBM Corp., Released 2012. IBM SPSS Statistics for windows, version 21.0. Armonk, NY: IBM Corp. USA). All the values were expressed as mean ± standard deviation and t-square test was used to assess the association between two parameters. Pearson's coefficient of correlation “r” was used to assess correlation. P < 0.05 was considered statistically significant.
| Results|| |
Serum and salivary SA and serum TP were statistically significantly higher in OSCC patients as compared to controls. However, there was no statistically significant difference between other parameters analysed [Table 2]. Comparison of serum and salivary SA, protein, and sugar with clinical stages of OSCC showed no statistically significant correlation [Table 3].
|Table 2: Comparison of serum and salivary sialic acid, protein, and sugar among oral squamous cell carcinoma and controls study patients (n=138)|
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|Table 3: Comparison of serum and salivary sialic acid, protein, and sugar with clinical staging of oral squamous cell carcinoma|
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Correlation of serum and salivary SA, protein, and sugar with histopathological grades of OSCC showed the serum SA was significantly higher in PDSCC as compared to moderately differentiated squamous cell carcinoma (MDSCC) and well-differentiated squamous cell carcinoma (WDSCC). Furthermore, the salivary SA was significantly higher in PDSCC as compared to MDSCC [Table 4]. There was a significant correlation in serum and salivary sugar in the controls, whereas it was not significant in OSCC [Table 5].
|Table 4: Comparison of serum and salivary sialic acid, protein, and sugar with histopathological grades of oral squamous cell carcinoma|
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|Table 5: Correlation of serum and salivary sialic acid, protein, and sugar in oral squamous cell carcinoma study patients and controls|
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| Discussion|| |
The aim of the present study was to analyze serum and salivary l SA, protein, and sugar in OSCC patients and correlate these parameters with those of healthy controls and with different clinical stages and histopathological grades of OSCC. In the present study, the serum SA level was significantly higher in OSCC patients as compared to controls, similar to the observations of several other authors.,,,,,, It has been demonstrated that SA increases at the tumor cell surface, so the increase in their serum levels may be related to their increased release through increased turnover, secretion, and shedding.,,,,,, The mean value of serum protein too was significantly higher in OSCC patients than in controls. The serum sugar level also was higher in OSCC patients than in controls, and it was statistically significant although in the year 2013 similar study done by Dhakar et al., stated higher serum sugar level in OSCC patients than in control but was not statistically significant. No further research is reported in the literature (Pub med central 2013–2020) estimating serum protein and sugar in OSCC, and hence the results of our study can not be compared with any.
The mean value of salivary SA was significantly higher in OSCC as compared to the controls in the present study. Similar results were also observed by Dhakar et al., Koç et al., and Sanjay et al.,, This may be explained on the basis that some of the tumor-associated carbohydrate changes are not restricted only to the cell surface membranes but may also occur due to secreted mucin. Salivary protein did not show any significant increase in OSCC as compared to the controls similar to the study done by Dhakar et al. While Krasteva et al.,, and Sanjay et al. showed increased values in OSCC as compared to controls. The source and relevance of this discrepancy are unknown; these salivary proteins are nonspecific, and their exact significance and function in cancer are unidentified. Dhakar et al., in their study, found no significant difference in OSCC patients as compared to the controls in salivary sugar levels which is found similar to our study. However, Sanjay et al., reported a significantly higher sugar level in OSCC patients than in controls. The reason for this difference in observations cannot be exactly figured out but may be due to variations in the methodology of studies.
In the present study, the mean value of serum SA, protein, and sugar show a significant correlation with the clinical stages of OSCC, while Xing et al., and Rajpura et al., found a significant correlation only between serum SA and clinical stages of OSCC. They explained that the elevations in the levels of SA appeared to reflect tumor burden and metastasis. However Dhakar et al., in their study showed no significant correlation with the clinical stages and among the mean value of serum SA, protein, and sugar. The mean value of salivary SA correlated well with the clinical stages of OSCC; similar to the study done by Dhakar et al.
The mean value of serum and salivary SA was significantly higher in PDSCC followed by MDSCC and WDSCC and this finding for serum SA is consistent with the results obtained by Dhakar et al., and Rao et al., This may be attributed to tumor differentiation and increased metastasis-related shedding of malignant cells into the bloodstream. However, Rajpura et al., did not find any significant correlation between these parameters. Dhakar et al. in their study showed that mean values of serum and salivary protein and sugar did not correlate with the histopathological grades of differentiation which was not inconsistent with our study results.
The serum SA, l protein, and sugar did not show any significant correlation with salivary SA, protein, and sugar in OSCC. In controls, the serum SA and protein showed a negative correlation with salivary SA and protein which was not significant and it is found similar to a study done by Dhakar et al., in the year 2013.
| Conclusions|| |
This research validates previous findings of the accuracy of serum SA as a cancer predictor. Furthermore, since the procedure is simple and nondestructive, salivary SA could be used for early detection as a diagnostic cancer biomarker as well as to predict the prognosis of malignancy. However, further research with a large sample size is needed to assess the accuracy of these parameters as specific tumor markers.
The authors would like to acknowledge the Department of Oral Pathology and Department of Biochemistry for all help throughout the study.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Joshi M, Patil R. Estimation and comparative study of serum total sialic acid levels as tumor markers in oral cancer and precancer. J Cancer Res Ther 2010;6:263-6.
Rastogi P. Emergence of cancer stem cells in head and neck squamous cell carcinoma: A therapeutic insight with literature review. Dent Res J (Isfahan) 2012;9:239-44.
Berlin NI. Tumor markers in cancer prevention and detection. Cancer 1981;47:1151-3.
Chittemsetti S, Manchikatla PK, Guttikonda V. Estimation of serum sialic acid in oral submucous fibrosis and oral squamous cell carcinoma. J Oral Maxillofac Pathol 2019;23:156.
] [Full text]
Erbil KM, Jones JD, Klee GG. Use and limitations of serum total and lipid-bound sialic acid concentrations as markers for colorectal cancer. Cancer 1985;55:404-9.
Mishra A, Verma M. Cancer biomarkers: Are we ready for the prime time? Cancers (Basel) 2010;2:190-208.
Patel PS, Shah MH, Jha FP, Raval GN, Rawal RM, Patel MM, et al
. Alterations in plasma lipid profile patterns in head and neck cancer and oral precancerous conditions. Indian J Cancer 2004;41:25-31.
] [Full text]
Hangloo VK, Kaul I, Zargar HU. Serum sialic acid levels in healthy individuals. J Postgrad Med 1990;36:140-2.
] [Full text]
Dabelsteen E, Clausen H, Mandel U. Aberrant glycosylation in oral malignant and premalignant lesions. J Oral Pathol Med 1991;20:361-8.
Dhakar N, Astekar M, Jain M, Saawarn S, Saawarn N. Total sialic acid, total protein and total sugar levels in serum and saliva of oral squamous cell carcinoma patients: A case control study. Dent Res J (Isfahan) 2013;10:343-7.
Carew JF, Singh B, Shah JP. Workup and staging. In: Shah JP, Johnson NW, Batsakis JG, editors. Oral Cancer. London: Martin Dunitz Publication; 2003. p. 205-6.
Anneroth G, Batsakis J, Luna M. Review of the literature and a recommended system of malignancy grading in oral squamous cell carcinomas. Scand J Dent Res 1987;95:229-49.
Skoza L, Mohos S. Stable thiobarbituric acid chromophore with dimethyl sulphoxide. Application to sialic acid assay in analytical de-O-acetylation. Biochem J 1976;159:457-62.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
Xing RD, Wang ZS, Li CQ, Tang QY, Jiang CB, Zhang YZ. Total sialic acid as a tumor marker for oral cancer. Int J Biol Markers 1994;9:239-42.
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.
Rawal RM, Patel PS, Patel BP, Raval GN, Patel MM, Bhatavdekar JM, et al
. Evaluation of glycoprotein constituents in head and neck cancer patients undergoing radiotherapy. Head Neck 1999;21:192-7.
Wilma Delphine Silvia CR, Vasudevan DM, Prabhu KS. Evaluation of serum glycoproteins in oral carcinoma. Indian J Clin Biochem 2001;16:113-5.
Goodarzi MT, Shafiei M, Nomani H, Shahriarahmadi A. Relationship between total and lipid-bound serum sialic acid and some tumor markers. Iranian J Med Sci 2005;30:124-7.
Rajpura KB, Patel PS, Chawda JG, Shah RM. Clinical significance of total and lipid bound sialic acid levels in oral pre-cancerous conditions and oral cancer. J Oral Pathol Med 2005;34:263-7.
Nigam PK, Narain VS, Kumar A. Sialic acid in cardiovascular diseases. Indian J Clin Biochem 2006;21:54-61.
Silver HK, Rangel DM, Morton DL. Serum sialic acid elevations in malignant melanoma patients. Cancer 1978;41:1497-9.
Koç L, Yarat A, Emekli N, Serdengeçti S, Berkarda B. Salivary sialic acid and cancer. J Marmara Univ Dent Fac 1996;2:523-6.
Sanjay PR, Hallikeri K, Shivashankara AR. Evaluation of salivary sialic acid, total protein, and total sugar in oral cancer: A preliminary report. Indian J Dent Res 2008;19:288-91.
] [Full text]
Krasteva A, Aleksiev E, Ivanova A, Altankova I, Bocheva T, Stanimirov P, et al
. Salivary components of treated cancer patients and patients with precancerous lesions. J IMAB 2008;2:41-4.
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]