ORIGINAL RESEARCH PAPER Volume-7 | Issue-4 | April-2018 | ISSN No 2277 - 8179 | IF : 4.758 | IC Value : 93.

98

INTERNATIONAL JOURNAL OF SCIENTIFIC RESEARCH

“ALTERATION OF BETA 2 MICROGLOBULIN LEVEL IN ORAL SQUAMOUS CELL

CARCINOMA AND POTENTIAL MALIGNENET DISORDER”

Dental Science
Associate professor, Department of Oral Medicine & Radiology Rishiraj College of
Dr. Arpit Jain Dental Sciences, Bhopal (M.P)
Professor & Head, Department of Oral Medicine & Radiology Government Dental
Dr. Guruprasad R College, Shimla, (J&K)
Associate Professor, Department of Prosthodontics, Crown& Bridge Rishiraj College of
Dr. Shilpa Jain* Dental Sciences, Bhopal (M.P) *Corresponding Author
Mr. Feroz Naem Scientist, Central Research Laboratory, People's University, Bhopal (M.P)
Khan
Dr. Brajendra Senior Lecturer, Department of Prosthodontics, Rishiraj College of Dental Sciences,
Singh Tomar Bhopal (M.P)
ABSTRACT
Background and Objective: Tumor markers are substances, which change quantitatively in serum during tumor development. One such tumor
marker is β2 microglobulin. It (β2 m) is a 11,800 kD protein of 100 amino acids associated with cell membrane of all nucleated cells as the small
subunit of the MHC class I molecule. β2 m is present in small amounts in free form in the serum, cerebrospinal fluid and urine of normal people and
to a much greater degree in patients with malignancies. The purpose of this study was to assess the serum levels of β2 microglobulin in oral
squamous cell carcinoma and potential malignant disorder and to evaluate the possible role of β2 microglobulin as a biochemical parameter in the
diagnosis of oral cancer.
Method: Serum β2 microglobulin levels were evaluated using ELISA in 50 patients with oral squamous cell carcinoma, 60 patients with potential
malignant disorder (30 Oral Submucous Fibrosis and 30 Leukoplakia), and 50 age-& sex matched disease-free controls.
Results: It was observed that there was a significant increase in serum β2 microglobulin levels in oral squamous cell carcinoma patients as
compared to potential malignant disorder and controls. Also it was found that significant increase in serum β2 microglobulin levels in oral
squamous cell carcinoma patients as compared to leukoplakia but not significant increase in serum β2 microglobulin levels in oral squamous cell
carcinoma patients as compared to Oral Submucous Fibrosis. Significant increase in serum β2 microglobulin levels in Oral Submucous Fibrosis
patients as compared to Leucoplakia. Although, serum β2 microglobulin levels were increased in oral leukoplakia and Oral Submucous Fibrosis
compared to controls, it was found to be statistically insignificant.
Conclusion: From these results, it seems that evaluation of serum β2 microglobulin levels may be useful as one of the battery of tests in assessment
of oral carcinoma and leukoplakia.
KEYWORDS
Serum β2 microglobulin; Oral squamous cell carcinoma; leukoplakia; ELISA.

INTRODUCTION: protein and enzymes (LDH) etc. In addition to the markers already
Cancer constitutes a major health problem in developing countries, studied, several tumor makers with clinical promise need further
representing one of the leading causes of death. Although oral cancer evaluation.5 Two such tumor markers are beta 2 microglobulin and
represents 2– 4% of the malignancies in the West, it accounts for Sialic acid in serum.
almost 40% of all cancers in the Indian subcontinent.1 Squamous cell
carcinoma accounts for 92% of all malignancies of Head and Neck Beta-2-microglobulin (Β2M) is a protein of low molecular weight
region.2 (11,800 daltons). It was first isolated from urine in patients with
Wilson's disease in 1968.6 It was found on the cell membrane of all
A significant proportion of oral squamous cell carcinomas (OSCC) nucleated cells and platelets and it forms the light chain moiety of the
develop from premalignant lesions such as leukoplakia and conditions major histocompatibility antigens. Cell membrane turnover is the
such as oral submucous fibrosis.3 Precancerous lesion is defined as principle source of Β2M in blood, plasma and body fluids.7 Elevated
“morphologically altered tissue in which oral cancer is more likely to serum levels has been found to be associated with increasing age8,
occur than in its apparently normal counterpart”, e.g. leukoplakia, relation to immune system6 and a variety of malignancies and appears
erythroplakia, actinic cheilitis etc. Precancerous condition is defined to be a reflection of tumor load in patients with myeloma9, bronchial
as a “generalized state associated with significantly increased risk of carcinoma 10 , breast cancer 11 , nasopharyngeal carcinoma 7 and
cancer”, e.g. Oral Submucous Fibrosis, Plummer Vinson syndrome, squamous cell carcinoma of the head and neck12.
Lichen Planus, Xeroderma Pigmentosum, Dyskeratosis Congenita,
etc.4 The development of oral cancer is a multistep process arising from The present study is an attempt to correlate the serum levels of beta 2
pre-existing potentially malignant lesions and conditions. microglobulin in oral precancer and oral squamous cell carcinoma and
Leukoplakia and OSMF are the most common precancers representing to evaluate the role of the same as a biochemical parameter for
85% of such entities. screening purposes.

Tumor markers are substances that are produced either by the tumor MATERIAL AND METHOD:
itself or by the body in response to the presence of cancer or certain Patients and controls: Serum was obtained from 50 untreated,
benign (noncancerous) conditions that can aid in the diagnosis of clinically evident oral cancer patients, proved by clinical and
cancer and in the assessment of tumor burden. Tumor markers can histopathological evidence: 60 patients with potential malignant
often be detected in higher than normal amounts in the blood, urine, or disorder (30 leukoplakia and 30 OSMF) but no evidence of invasion:
body tissues of patients with certain types of cancer. Estimation of and 50 age and sex matched controls.
tumor marker level can be useful when used along with radiographs or
other tests in the detection and diagnosis of certain types of cancer.5 Subjects who had taken treatment for any Precancerous lesions/
conditions or Oral cancer and Subject suffer from any systemic
In the carcinomas of oral cavity, various serum markers have been diseases like diabetes, cardiac diseases, renal diseases, liver diseases
studied: these include oncofetal proteins (alpha-fetoprotein, CEA), B- and other malignancies and those who are taking antioxidants/

International Journal of Scientific Research 559

Volume-7 | Issue-4 | April-2018 ISSN No 2277 - 8179 | IF : 4.758 | IC Value : 93.98

multivitamin preparations were excluded from the study. The level of Β2M in oral cancer patients was increased when compared
with controls which was in accordance with few other studies reported
Patient further divided into 3 groups Group A: 50 healthy individuals, in the literature.7, 12, 13-17 We also found the increased level of Β2M in
38 males and 12 females. Group B: 60 potential malignant disorder, 51 different stages (stage II &III) of oral cancer. The mechanism of
males and 9 females. Group B further divided into two groups, Group increase in Β2M levels in malignancies is not known but various
Ba: 30 patients with Luekoplakia and Group Bb: 30 patients having possible hypotheses have been put forward. The Β2M is a cell
OSMF. Group C: 50 oral carcinoma patients, 40 males and 10 females. membrane constituent along with the HLA chain, so an accelerated
membrane turnover or accelerated cell division could increase the
METHODS OF SAMPLE COLLECTION shedding of Β2M. The ability of the carcinoma cells to produce a
Ethical clearance was obtained from the institute and the hospital. All higher concentration of Β2M than the non-neoplastic cells may be due
the patients fulfilling the above criteria were informed about the study to either active synthesis or increased cell breakdown or both.14, 15 In
being conducted and only those who agreed were enrolled in the study. many neoplasms, especially those of epithelial origin, a decrease or
All the enrolled subjects were then interviewed using clinical lack of expression of HLA-I particles was reported. The weakened
examination tools and recorded in a case history Performa. expression of HLA-I complex on the neoplastic cells may lead to
increased level of Β2M in the blood serum. Recent studies explained
After obtaining consent from the patient, 5 ml of venous blood samples this phenomenon as due to an imbalance of light chain and heavy chain
were collected by venipuncture of the median cubital vein under of HLA-I complex and relative over -production of Β2M .15 Most
aseptic precautions. The blood samples were allowed to clot for 1 hour frequently quoted hypothesis for high levels of Β2M in neoplastic
and then centrifuged at 3000 rpm for 10 minutes to provide serum. This diseases explains this phenomenon with mono or polyclonal activation
serum was preserved in a frozen state at -20 degree until the analysis. of lymphocytes, destruction of MHC I (Major histocompatibility
complex) particles, and increased cellular transformation into
Beta 2 microglobulin was estimated by using ELISA method for neoplastic cells which could lead to higher concentration of protein
which Kit was obtained from DRG International, Inc., USA Β2M . Certain studies proposed that the systemic immunosuppression
observed in oral cancer patients is due to the decreased functional
PRINCIPLE OF THE ASSAY activity of peripheral blood monocytes which is reflected by way of
The β-2-Microglobulin ELISA test is based on the principle of a solid decreased phagocytic process.18 The latest studies showed that a high
phase enzyme-linked immunosorbent assay (ELISA). The concentration of free Β2M may have a negative influence on the
concentration of β-2-Microglobulin is directly proportional to the immunological system by decreasing the expression of MHC-I
color intensity of the test sample. Absorbance is measured particles and indirectly by increasing the levels of cytokines: IL-6, IL-
spectrophotometrically at 450nm. 10, which accelerate the development of neoplasms.17,19

Thus the observations obtained by these methods were tabulated and The Β2M level is also significantly increased in Oral Precancer group
statistically analyzed By STUDENT 't' TEST and Tukey's HSD when compared with controls. Similar findings were also observed in
test. other studies.12, 14 In the present study the level of Β2M in leukoplakia
patients were significantly increased when compared with controls but
RESULT: it differed from one study conducted in Trivandrum which shows no
The mean serum Β2M levels in group A was 2.57 ± 0.10 µg/ml. It was change in the mean value of Β2M in leukoplakia 14. It could be
elevated when compared to group C in which it was 0.89 ± 0.02 µg/ml. attributed to geographic variations and/or selection criteria
It is statistically significant (p value <0.0001). {Table 1, Graph 1} The differences. In this study we also observed that the level of Β2M was
mean serum Β2M levels in Group B was 2.23 ± 0.33 µg/ml, it was slightly elevated in speckled leukoplakia as compared to
significantly elevated when compared to Group C in which it was 0.89 homogeneous leukoplakia but it was not statistically significant; the
± 0.02 µg/ml. It was statistically significant (p value <0.0001). {Table reason behind this could be that speckled leukoplakia had more
1, Graph 1} The mean serum Β2M levels in Group A was 2.57 ± 0.10 malignant potential as compared to homogenous leukoplakia.4 The
µg/ml. It was significantly elevated when compared to Group B (2.23 ± Β2M level was also significantly increased in OSMF patients when
0.33 µg/ml). It was statistically significant (<0.0001). {Table 1, compared with controls. Similar finding was also observed in one
Graph1} study reported in the literature.14 The level of Β2M was also marginally
elevated in different OSMF stages but it was not statistically
The mean serum Β2M levels in Group A was 2.57 ± 0.10 µg/ml. It was significant.
elevated when compared to Group Ba (1.89 ± 0.03 µg/ml) which was
statistically significant (p value <0.0001). {Table 2, Graph 2} Increased Β2M levels were observed in Oral Leukoplakia, which was
found to be significant. The increased levels of Β2M in Oral
The mean serum Β2M levels in group A was 2.57 ± 0.10 µg/ml. It was Leukoplakia patient's serum may be due to increased production or
elevated when compared to Group Bb (2.56 ± 0.04 µg/ml) which was impaired excretion.14 The reason for increased level of Β2M in OSMF
not statistically significant. {Table 3, Graph 3} is unknown but it has been suggested that cell mediated and humoral
immune responses have been reported to be altered in patients with
The mean serum Β2M levels in Group Ba was elevated (1.89 ± 0.03 OSMF and OSMF also has high rate of malignant transformation. It
µg/ml) when compared to Group C (0.89 ± 0.02 µg/ml) which was has been proposed that OSMF may be an intermediary stage in
statistically significant (p value <0.0001). {Table 2, Graph 2} malignant transformation.14. This can be the plausible explanation for
the reason behind the increased level of B2M in OSMF patient.
The mean serum Β2M levels in Group Bb was 2.56 ± 0.04 µg/ml. It
was elevated when compared to Group C (0.89 ± 0.02 µg/ml), which In present study the level of Β2M in oral cancer patient was
was statistically significant (p value <0.0001). {Table 3, Graph 3} significantly increased when compared with leukoplakia patients
which is comparable to a study reported in the literature.20 The Β2M
The mean serum Β2M levels in Group Bb 2.56 ± 0.04 µg/ml. It was level was not significantly increased in oral cancer patients when
elevated when compared to Group Ba (1.89 ± 0.03 µg/ml), which was compared with OSMF patients. Similar finding was also found in a
statistically significant (p value <0.0001). {Table 4, Graph 4} study reported in the literature.14

DISCUSSION: In present study, the level of Β2M in OSMF patients were significantly
Various changes occur in the body in presence of any type of cancer increased when compared with leukoplakia patients which was
because the tumor cells produce certain types of chemical mediators comparable to a study reported in the literature.14
into blood like oncofetal proteins (alpha-fetoprotein,
carcinoembryonic antigen), B-protein, enzymes (e.g. Lactate CONCLUSION:
Dehydrogenase), Β2M and sialic acid etc. In the presence of any kind Identification of reliable biological tumor markers or substance
of tumor, the levels of these substances will change. In the present associated with neoplasia that can be used for the detection, staging
study, the levels of Β2M and sialic acid in oral cancer and precancer and evaluation has been the goal of many investigators. On the basis of
patients were estimated and compared with the levels of normal present study we concluded that the levels of β 2 microglobulin was
healthy, deleterious habit free individuals. increased in oral cancer and precancer, however further
comprehensive studies involving large sample size are required to

560 International Journal of Scientific Research

Volume-7 | Issue-4 | April-2018 ISSN No 2277 - 8179 | IF : 4.758 | IC Value : 93.98

confirm the clinical usefulness of serum β 2 microglobulin as a

biochemical parameter.

TABLE: 1 BETA 2 MICROGLOBULIN LEVELS IN STUDY

AND CONTROLS GROUPS
PARAMETERS GROUP N Mean ± SD p-Value Result
Β2M (mg/ml) Group C 50 0.89 ± 0.02 <0.0001 Significant
Group A 50 2.57 ± 0.11
Group B 60 2.23 ± 0.34
TABLE: 2 BETA 2 MICROGLOBULIN LEVELS IN GROUP A,
GROUP C AND LEUKOPLAKIA
PARAMETERS GROUP N Mean ± SD p-Value Result
Β2M (mg/ml) Group C 50 0.89 ± 0.02 <0.0001 Significant GRAPH: 4 MEAN VALUE OF BETA 2 MICROGLOBULIN IN
OSMF AND LEUKOPLAKIA
Group A 50 2.57 ± 0.11
LEUKOPLAKIA 30 1.89 ± 0.03 REFERENCES:
TABLE: 3 BETA 2 MICROGLOBULIN LEVELS IN GROUP A, 1. Mehrotra R, Singh M, Kumar D, Pande AN, Gupta RK, Sinha US. Age specific
incidence rate and pathological spectrum or oral cancer in Allahabad. Ind J Med Sci.
GROUP C AND OSMF 2003;57:400–04.
PARAMETERS GROUP N Mean ± SD p-Value Result 2. Mishra A. Head and neck cancer in India – review of practices prevention policy. Oral
Diseases. 2009;15:454-65.
Β2M (mg/ml) Group C 50 0.89 ± 0.02 <0.0001 Significant 3. S. Humayun and V. Ram Prasad Expression of p53 protein and ki-67 antigen in oral
Group A 50 2.57 ± 0.11 premalignant lesions and oral squamous cell carcinomas: An immunohistochemical
study Natl J Maxillofac Surg. 2011 Jan-Jun; 2(1): 38–46.
OSMF 30 2.57 ± 0.04 4. R Rajendran. Benign and Malignant Tumors of the oral cavity in: R Rajendran, B
Sivapathasundhram (eds.) Shafer’s text book of Oral Pathology 5th edi. New Delhi,
TABLE: 4 BETA 2 MICROGLOBULIN LEVELS IN OSMF AND India: Elsevier publication 2005: 122,123,136.
LEUKOPLAKIA 5. Reddy SB, Reddy MB, Shyam NDVN. Tumour Markers in Oral Neoplasia. IJDA.2010;
2(1):103-14.
PARAMETERS GROUP Mean n p-Value Result 6. GROVES ML and GREENBERG R. β2-Microglobulin and Its Relationship to the
± SD Immune System. J Dairy Sci. 1982;65:317-25.
Β2M (mg/ml) OSMF 2.57 ± 30 <0.0001 Significant 7. Shiu W, Leung SF, Leung WT, Ho S & Tao M. Expression of beta-2-microglobulin by
nasopharyngeal carcinoma Br. J. Cancer (1992), 66, 555-57.
0.04 8. Merrett .J, Atkinson P, Burr M. and Merett.T.G. Circulating levels of β2- microglobulin
LEUKOPLAKIA 1.89 ± 30 in the age group of patients over 70s. clinical chimica acta. 1980;104:119-23.
0.03 9. Norfolk D, Child JA, Coopert EH, Kerruisht S and Ward AM. Serum β2-microglobulin
in myelomatosis: potential value in stratification and monitoring. Br J Cancer. 1980;
42:273-78.
10. Hallgren R, Nou E, Lundqvist G. Serum β-2-Microglobulin in Patients with Bronchial
Carcinoma and Controls. Cancer. 1980; 45:780-785.
11. Papaioannou D, Geggie P, and Klassen J. Study of serum β2-microglobulin levels in
breast cancer patients. clinical chimica acta.1979;79:135-43.
12. SCULLY C. Serum β 2 microglobulin in oral malignancy and premalignancy. Journal of
Oral Pathology 1981; 10: 354-57.
13. Shabana AHM. Expression of beta-2-microglobulin by normal, benign and malignant
oral epithelia. The Saudi Dental Journal. 1991; 3(3):92-98.
14. Anil S, Beena VT, Nair RG, and Vijayakumar T. Evaluation of serum 32-microglobulin
in premalignant and malignant lesions of the oral cavity. Oral Surg Oral Med Oral Patho
Oral Radiol Endod 1995;79:750-2.
15. Silvia C.R.W.D, Vasudevan D.M, and Sudhakar Prabhu K.: Alteration of serum β2-
Microglobulin in oral carcinoma. Ind J of Clin Biochem 2002; 17(2):104-107.
16. Chen C-H, et al. Overexpression of BETA 2-microglobulin is associated with poor
survival in patients with oral cavity squamous cell carcinoma and contribution to oral
cancer cell migration and invasion. British Journal of Cancer. 2008; 99: 1453 – 61.
GRAPH: 1 MEAN VALUE OF BETA 2 MICRIGLOBULIN IN 17. Kadam CY, Katkam RV, Suryakar AN, Kumbar KM, Kadam DP. Biochemical markers
STUDY AND CONTROLS GROUPS in oral cancer. Biomedical Research 2011; 22 (1): 76-80.
18. Reshma K, Bharathi B. Phagocytosis: A marker of decreased immune response in
radiation treated oral cancers. Biomedical Research. 2009; 20 (1): 75-77.
19. Xie J, Wang Y, Freeman III M E., Barlogie B and Yi Q. β2-Microglobulin as a negative
regulator of the immune system: high concentrations of the protein inhibit in vitro
generation of functional dendritic cells. BLOOD. 2003;101(10):4005-12.
20. Greipp PR, Katzmann JA, O’Fallon WM, and Kyle RA. Value of β 2-Microglobulin
Level and Plasma Cell Labeling Indices as Prognostic Factors in Patients with newly
Diagnosed Myeloma. Blood. 1988 72: 219-23.

GRAPH: 2 MEAN VALUE OF BETA 2 MICROGLOBULIN IN

GROUP A, GROUP C AND LEUKOPLAKIA

GRAPH: 3 MEAN VALUE OF BETA 2 MICROGLOBULIN IN

GROUP A, GROUP C AND OSMF
International Journal of Scientific Research 561