|Year : 2005 | Volume
| Issue : 1 | Page : 16-19
An estimation of serum β-2 microglobulin level in premalignant lesions / conditions and oral squamous cell carcinoma: A clinicopathological study
N Vaishali, JV Tupkari
Department of Oral and Maxillo Facial Pathology, Govt. Dental College and Hospital, Aurangabad - 431 041, India
J V Tupkari
Department of Oral and Maxillo Facial Pathology, Govt. Dental College and Hospital, Aurangabad 431 001
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Recently biological tumour markers have been introduced into the clinical diagnosis of malignant lesions. Tumour markers are the substances, which quantitatively change in serum during tumour development. One such tumour marker is "Beta 2 microglobulin". In the present study, an attempt was made to correlate the level of serum Beta 2 microglobulin with premaIinant lesions / conditions and oral squamous cell carcinoma. This study has been carried out to evaluate the role of Beta 2 microglobulin as biochemical parameter in oral premaligant lesions conditions and oral squamous cell carcinoma. The progressively increased serum β-2 microglobulin level has positive correlation with the histologic grading of oral squamous cell carcinoma. Also increased level of β-2 microlobulin was observed in oral premalignant lesions and conditions
Keywords: β-2 microglobulin, IRMA
|How to cite this article:|
Vaishali N, Tupkari J V. An estimation of serum β-2 microglobulin level in premalignant lesions / conditions and oral squamous cell carcinoma: A clinicopathological study. J Oral Maxillofac Pathol 2005;9:16-9
|How to cite this URL:|
Vaishali N, Tupkari J V. An estimation of serum β-2 microglobulin level in premalignant lesions / conditions and oral squamous cell carcinoma: A clinicopathological study. J Oral Maxillofac Pathol [serial online] 2005 [cited 2020 Sep 27];9:16-9. Available from: http://www.jomfp.in/text.asp?2005/9/1/16/39053
| Introduction|| |
Oral cancer is highly prevalent in Indian population, primary associated with various habits. A close correlation is postulated between tobacco chewing habit and the high incidence of oral precancer and cancer  . Recently, there have been a number of scientific approaches to the problem of precancerous lesions, with aim to establish fundamental biochemical basis of understanding  . The goal of such methods is to find a reliable indicator of the biological potential of precancerous and cancerous lesions. The quench for the early detection of the lesion has made it very important for the dental professionals to maintain high levels of diagnostic methods to assure the diagnosis  .
Currently, tumour markers have been introduced for the early detection of the lesion. These markers have wide range of potential applications: for screening purpose, diagnosis, prognosis, and monitoring the response to treatment. Estimation of such markers permit selection of the most appropriate treatment for individuals. The search For "ideal tumour marker" has become a major goal in oral pathology  . Identification of such ideal tumour marker can offer an exciting opportunity for early detection of the lesion. Therefore study of tumour marker has become focal point of research in oncology.
In the oral cavity, various markers have been studied: these include oncofoetal protein, (a- fetoprotein: CEA), other proteins B-Protein, β-2 microglobulin), and enzymes (LDH). One such marker is β-2 microglobulin. Crispian Scully was the first to assess the potential use of β-2 microglobulin as a marker in oral premalignant lesions  .
In the present study, an attempt was made to correlate the level of β-2 microglobulin with premalignant lesions conditions and oral squamous cell carcinoma. This study has been carried out to evaluate the role of β-2 microglobulin as a biochemical parameter in oral permalignant lesions / conditions and oral squamous cell carcinoma.
| Purpose of the Study|| |
- To study the clinical and histological features in premalignant lesions / conditions and oral squamouscell carcinoma.
- To estimate the scrum β-2 microglobulin level.
- To compare serum β-2 microglobulin level in premalignant lesions / conditions and malignant lesions.
- To determine the correlation clinical and histopathological degree with β-2 microglobulin level.
- To predict the role of β-2 microglobulin as a biochemical parameter for the diagnosis and prognosis of oral squamous cell carcinoma.
Estimation of β-2-microglobuIin levels
In the present study, commercially available Coal-A-Count β-2 microglobulin IRMA kit was used for the quantitative measurement of β-2 rnicroglobulin in the serum [Figure - 1].
Principle of the procedure
Coat-A-count β-2 rnicroglobulin IRMA is a solid phase immunoradiometric assay based on monoclonal and poly clonal anti β-2 microglobulin antibodies. One 125-1 labeled anti β-2 microglobulin monoclonal antibody in liquid phase and one polyclonal anti β-2 microglobulin antibody immubilieed of the wall of polysteene tube. β-2 rnicroglobulin is captured between polyclonal anti β-2 microglobulin tracers. The unbound 125 I labeled anti β2-microglobulin antibody is removed by decanting the reaction. The β-2 microglobulin concentration is directly proportional to radioactivity present in the test tube.
| Material and Methods|| |
The study comprised of 80 subjects grouped as Group I - Control (20 subjects), Group II - Premalignant lesions / conditions (30 subjects), and Group III - Oral squamous cell carcinoma (30 subjects).
Thorough screening and clinical examination of each patient was done. Biopsy was performed fir the confirmation of diagnosis. 3-4 ml of I .V blood was collected and transferred to test tube and scrum was separated out.
Estimation of β-2 rnicroglobulin level in serum was done by "Immunoradiometric Assay" [Figure - 1] IRMA is an excess reagent technique in which excess radiolabeled anti body is al lowed to react with the analyte (Ag) from the sample.
Immunometric Assay Procedure
- All serum samples are to be diluted 1 in 21 with zero calibrator before assay. In the present study, 20 ml of each serum sample was diluted with 0.4 ml of zero calibrator.
- β-2 microglobulin Ab coated tubes were labeled according to calibrators such as (i) 10, 25. 57, 111, 268, 567, 1040 mg/ml and study group. (ii) Group 1: control (iii) Group II: Premalignant lesions / conditions, (iv) Group III: Oral squamous cell carcinoma.
- With the help of micropipette, 20 ml of each calibrator and all diluted serum samples from each group were pipetted in Ah coated labeled tubes.
- 0.4 ml β-2 microglobulin assay buffer solution was added to each tube.
- All tubes were shaken for 30 minutes on rack shaken.
- Afters shaking, the tubes were decanted thoroughly.
- 4 m of buffer wash solution was again added to each tube. After 1- 2 min, the tubes decant thoroughly.
- 0.4 ml of 125-1 β-2 microglobulin Ab (Tracer) was added 30 each tube of sample.
- After addition of tracer, all tubes were shaken for 30 minutes
- After shaking all tubes were decant thoroughly. The unbound 1-125 labeled anti β-2 microglobulin antibody was removed by this procedure.
- Antigen bound fraction within latch tube was counted for 1 minute in gamma counter. Counts per minute of all samples from each group were recorded.
- The counts per minutes of all calibrators were plotted on logit log paper. The standard calibrator curve was plotted on the graph paper.
- The standard calibrator curve was microglobulin curve was estimated by interpolation on 125 calibrator curve.
The concentration obtained was multiplied by 21, Statistical analysis of β-2 microglobulin level is done by student`t'test.
| Result and Observation|| |
In the present study, the serum β-2 microglobulin level was estimated in the groups, according group I for control subjects. Group II for premalignant lesions / conditions and Group III for squamous cell carcinoma. The arithmetic mean of serurn β-2 microglobulin level of each group was calculated. The standard deviation of each parameter was calculated. Results are given in [Table - 1].
A progressive increase in the scrum β-2 microglobulin level was observed in Group II and Group III parameters [Table - 1].
The statistical difference between serum β-2 microglobulin level in control and oral premalignant lesions / conditions, control and oral squamous cell carcinoma were analysed. 1 [Table - 2] and [Table - 3].
As two independent samples were to he correlated, the student 't' test was applied. With the 't' value, probability p was calculated. Comparison of serum β-2 microglobulin in control vs. oral premalignant lesions / conditions, control VS oral squmaous cell carcinoma, for p value less than 0.005, the statistical difference between these two group was found to be highly significant.
| Discussion|| |
Oral cancer is highly prevalent in Indian population primarily associated with various habits. A close correlation is postulated between tobacco chewing habit and high incidence of oral precancer and cancer. In most instances, these patients seek medical attention only at an advanced stage, thereby leading to poor prognosis and postoperative disfigurement. The current mortality rate, attributed to oral cancer can be reduced greatly if early signs and symptoms are given an adequate attention. Therefore, the detection of the lesion at an early stage is of paramount prognostic importance  . Recently, tumour markers are receiving more attention in the early detection of the lesion. Various biological markers have ushered in the field of dentistry  .
In the present study, quantitative analysis of β-2 microglobulin was done using commercial kit. β-2 microglobulin is a low molecular weight protein (11800 dalton), chiefly present on the surface of nucleated cells, abundantly on lymphocytes and tumour cells. Crispian Sculls was the first to evaluate the role of β-2 microglobulin in premalignancy and malignancy  .
The mechanism of altered β-2 mnicroglobulin level is not yet clearly understood. Various possibilities for increased serum level of β-2 microglobulin Suggested are an increased cellular activity in malignancy being responsible for an increased release of β-2 microglobulin, β-2 microglobulin being a constituent of HLA antigen, cell membrane turnover or cell division could increase the shedding of β-2 microglobulin  .
The observed mean scrum β-2 microglobulin level was 1.17 mg./L in the control group, I-59 mg/L, in oral leukoplakia and oral submucous fibrosis, 1.77 mg/L in hyperkeratonic complex, which is slightly higher than hyperkeratotic simplex which is 1.40 mg/L [Table - 2]. This progressively increasing β-2 microglobulin level positively correlates with the degree of cellular atypia (G) suggesting that β-2 microglobulin level can serve as biochemical tool in assessing the malignant potential of premalignant lesions.
In 30 sections, which comprised of 25 well differentiated oral squamous cell carcinomas, mean serum β-2 microglobulin level was 2.2 mg/L while it was 2.4 mg/L in 5 cases of moderately differentiated oral squamous cell carcinoma. The significant increased β-2 microglobulin level relates with increased cellular activity. The disturbances in the cell surface is responsible for the excessive shedding of β-2 microglobulin. Increased serum β-2 microglobulin level correlates with the degree of dvsplasia i.e. histological grade.
The results of the present study are in agreement with the previous studies wherein Washif Manzar observed a definite increase in serum β-2 microglobulin level, that is. 2.5 mg/L  and S Anil et al noted the level of β-2 microglobulin in oral squamous cell carcinoma to be 2.19 mg/L, [Table - 3]  . In the present study, a definite increase in the level of β-2 microglobulin from control group to premalignant lesions / conditions and oral squamous cell carcinoma was statistically significant. These results are in agreement with the studies of Silivia and Prabhu  .
Thus estimation of β-2 microglobulin level may provide the biochemical stratification of the disease. This can act as a biochemical parameter in the diagnosis of the lesion.
| Summary and Conclusion|| |
In the present study, total 80 subjects were assessed for serum β-2 microglobulin level and the detailed clinical examination and relevant history of each patient was recorded thoroughly. The estimation of serum β-2 microglobulin was done by using commercial kit, Coat A Count β-2 microglobulin. The mean scrum β-2 micro globulin level was correlated with clinical and histopathological findings in PML and squamous cell carcinoma. The progressively increasing serum β-2 microglobulin levels have positive correlation with histologic grading of squamous cell carcinoma. Also increased level of β-2 microglobulin was observed in oral premalignant lesions and conditions.
Thus, the estimation of β-2 microglobulin levels is found to be specific and sensitive test for diagnosis and prognostic evaluation.
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[Figure - 1], [Figure - 2]
[Table - 1], [Table - 2], [Table - 3]