ORIGINAL RESEARCH       

Year : 2005  |  Volume : 9  |  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

Correspondence Address:
J V Tupkari
Department of Oral and Maxillo Facial Pathology, Govt. Dental College and Hospital,
Aurangabad 431 001 
India

Source of Support: None, Conflict of Interest: None

Check

DOI: 10.4103/0973-029X.39053

 
    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 2019 Jun 16];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 [4] . Recently, there have been a number of scientific approaches to the
problem of precancerous lesions, with aim to establish fundamental biochemical basis of
understanding [6] . 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 [2] .

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 [6] . 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 [5] .

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  

1. To study the clinical and histological features in premalignant lesions / conditions and
oral squamouscell carcinoma.
2. To estimate the scrum β-2 microglobulin level.
3. To compare serum β-2 microglobulin level in premalignant lesions / conditions and
malignant lesions.
4. To determine the correlation clinical and histopathological degree with β-2 microglobulin
level.
5. 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

 1. 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.
2. 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.
3. 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.
4. 0.4 ml β-2 microglobulin assay buffer solution was added to each tube.
5. All tubes were shaken for 30 minutes on rack shaken.
6. Afters shaking, the tubes were decanted thoroughly.
7. 4 m of buffer wash solution was again added to each tube. After 1- 2 min, the tubes
decant thoroughly.
8. 0.4 ml of 125-1 β-2 microglobulin Ab (Tracer) was added 30 each tube of sample.
9. After addition of tracer, all tubes were shaken for 30 minutes
10. After shaking all tubes were decant thoroughly. The unbound 1-125 labeled anti β-2
microglobulin antibody was removed by this procedure.
11. 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.
12. The counts per minutes of all calibrators were plotted on logit log paper. The standard
calibrator curve was plotted on the graph paper.
13. 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 [4] . Recently, tumour markers are receiving more
attention in the early detection of the lesion. Various biological markers have ushered in the field
of dentistry [12] .

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 [7] .

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 [1] .

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 [3] 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] [1] . 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 [9] .

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.

 
    References  

1. Anil S. Beena VT, Raj G. Nair et al: Evaluation of scrum β-2 microglobulin in premalignant
and malignant lesions of the oral cavity- Oral surgery, Oral Medicine, Oral pathology, Oral
Radiology, Endodontics 1995: Vol.79 (6): pp.750-52.       
2. Hays F Washif, Vijay Raghavan et al: Evaluation of β-2 globulin in oral cancer. Australian
Dental clinics of North America. June 1994; Vol,8(3).       
3. Manzar Washif, Vijay Raghavan et al: Evaluation of β-2 globulin in oral cancer. Australian
Dental Journal 1992; Vol.37, No.1: pp- 39-42       
4. Rao RA and Desai PB: Oral Cancer, professional Education Division. Tata Memorial
Hospital and Centre Bombay. 1991; l st edition.       
5. Rassekh Christopher, Johnson et al: Circulating markers in squamous cell carcinoma of
head and neck: A review- Oral Oncology. European journal of cancer 1994: Vol.308, No.1:
pp.23-28.       
6. Scully Gispian Burkhwdt A: Tissue markers of potentially malignant human oral epithelial
lesions. Journal of Oral Pathology Medicines 1993; Vol.22: 246-256.       
7. Scully Ca: serum β-2 microglobulin in oral malignancy and per malignancy. JOP 1981; Vol.
10: pp.354 - 357.       
8. Shafer W G, Hine MK, Levy BM: A textbook of oral pathology- 4th edition, WB Saunders
Publication 1993, Philadelphia. Pp.112-130.       
9. CR Wilma Delphine silivia, DM Vasudevan & K Sudhakar Prabhu: Alteration of serum β-2
microglobulin in oral carcinoma. Indian Journal of Clinical Biochemistry 2002: 17(2): 104-
107.       
10. Steward sell: Cancer markers. Comprehensive Textbook of Oncology, Williams and
Willkins publication, London, 1986; 1st edition.       
11. Tropes G Class, Logdbers Lennwet et al: β-2 microglobulin; A tumour marker of
gynaecologic cancer. American journal of Obstetrics and Gynaecology. 1981;Vol. 137 (6):
pp.743-744.       
12. Vinez Kort, S EVA et al: Diagnosis of head and neck carcinoma by means of immuno
marker. J Cranio maxillogical surgery. 1987: Vol .15: pp, 270-277.