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RA JIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA
ANNEXURE – II
Proforma for Registration of Subjects for Dissertation
1
Name and Address of Dr. Nandini Mohan. C.,
the Candidate
Postgraduate Student,
Dept. of Oral and Maxillofacial Pathology,
AECS Maaruti College of Dental Sciences and Research
Centre,
Bangalore – 560 076.
2
Name of the Institution
AECS Maaruti College of Dental Sciences and Research
Centre.
3
Course of Study and Master of Dental Surgery
Subject
4
Oral and Maxillofacial Pathology.
Date of Admission to June 2013
the Course
5
Title of the Topic
Expression of Human Chorionic Gonadotropin-β in
serum and tissue sections of preoperative oral squamous
cell carcinoma patients.
6. Brief resume of intended work:
Head and neck cancer is the sixth most common type of cancer in the world.
Histopathologically, Squamous Cell Carcinoma (SCC) is by far the most frequent
histological type of this disease. After curative treatment, about 50% of the patients
develop a recurrence and 80% of the relapses occur within the first 2 years of follow
up. About 50% of the patients with SCC of head and neck die from cancer with no
significant improvement during recent decades1.
In the management of patients with SCC of head and neck, treatment decisions
are still based on the TNM classification system, although T and N status do not
reliably predict the clinical outcome in individual cases. Thus, effective treatment
planning would be enhanced by identification of prognostic indicators that would
reflect the biological behavior of a tumor. The identification of a reliable circulating
tumor marker therefore has great potential. Although new potential prognostic
markers are continuously being introduced and evaluated in SCC of head and neck,
so far none has influenced standard treatment1.
Human Chorionic Gonadotropin (hCG) is a hormone belonging to the
glycoprotein family. It is initially secreted by the pre-implantation embryo and
subsequently by trophoblasts. It is responsible for rescue of the corpus luteum,
thereby sustaining progesterone secretion until placental progesterone synthesis
commences. hCG is a heterodimeric molecule consisting of an α-subunit
noncovalently associated with a hormone specific β-subunit. The α-subunit is
common to hCG, Luteinising hormone, Follicle stimulating hormone and Thyroid
stimulating hormone and contains 92 amino acids. The β-subunit of hCG confers
biological specificity and contains 145 amino acids2.
hCG is normally produced in significant amounts only during pregnancy. It is also
ectopically made by trophoblastic as well as non-trophoblastic(colon, prostate,
bladder, breast and lung) carcinomas3. β-hCG has therefore been proposed as a
cancer marker of broad utility.
In general, hCG and/or subunit synthesis is seen in poorly differentiated tumors. hCG
expression is prominent in metastatic cancers and is associated with poor prognosis4.
Survival time of patients with hCG expressing cervical, pancreatic and colorectal cancers
has been reported to be statistically shorter than those with hCG negative neoplasms. In
bladder cancer as well, the serum level of hCG is considered a prognostic indicator of
disease. The presence of hCG has been linked to chemo- and radio- resistance and the
molecule has been shown to increase invasiveness1.
Thus evaluation of β-hCG levels in patients with SCC of oral cavity has significant
diagnostic and prognostic value.
6.1 Need for the Study:
The aim of this study is to observe the expression of hCG-β in serum by ELISA and its
immunohistochemical expression in tissue sections of patients with oral squamous cell
carcinoma preoperatively and to quantify and compare the same.
6.2 Review of Literature:
Iles R K et al (1990) studied the expression of hCG-β by non-trophoblastic, nonendocrine, normal and malignant epithelial cells. They examined 83 different cell lines
derived mainly from common epithelial tumours. 32 of the cell lines were found to
secrete hCG like material into their culture media. Partial immunochemical
characterization showed that of these only choriocarcinoma and fetal tissue cell lines
produced intact hCG and α-subunit. The remaining 28 hCG expressing epithelial cell
lines, which are of mucosal origin, only secreted free β-subunit. Expression of free hCG-β
by non trophoblastic non endocrine cells would appear to be especially characteristic of
mucosal epithelia from the genitourinary and oral/respiratory tracts. It may also show
squamous cell-like properties.
Marcillac I et al (1992) studied free hCG-β subunit in serum of patients with gonadal
neoplasms and nongonadal neoplasms and concluded that detection of free hCG-β subunit
in serum of nonpregnant subjects was highly diagnostic of malignancy in general and
specifically defines a subgroup of aggressive nongonadal malignancies.
Sheaff M T et al (1996) studied the importance of immunohistological detection of hCGβ in localized prostatic adenocarcinoma with regard to prognosis and clinical applications.
They studied 80 cases of clinically localized adenocarcinoma of prostate and found that
hCG-β was detected in 12 cases. 9 of these patients were found to have metastases at
follow up and 11 were dead within 18 months. They thus concluded that the
demonstration of hCG-β in prostatic adenocarcinoma identifies a group of patients with
poor prognosis, irrespective of histological grade. This information is extremely valuable
in the subsequent clinical management of such patients.
Johan Hedstrom et al (1999) studied the concentration of free hCG-β subunit in serum
as a prognostic marker for squamous cell carcinoma of oral cavity and oropharynx. They
studied 59 cases of squamous cell carcinoma of oral cavity and oropharynx
preoperatively. Elevated preoperative hCG-β levels were observed in 8 patients and these
patients showed shorter recurrence free survival than those with normal hCG-β levels.
Risk for recurrent disease in patients with preoperatively elevated hCG-β was 3.6 folds
more than that of patients with normal hCG-β levels.
Bhalang et al (1999) conducted an immunohistochemical study of the expression of
hCG-β in Oral Squamous Cell Carcinomas (OSCC) in comparison with oral fibromas.
hCG-β immunoreactivity was identified in 29 of 45 cases of oscc. hCG-β
immunoreactivity could not be demonstrated in any of the oral fibromas. hCG-β staining
was positive in 5 of 15 cases of well differentiated OSCC, in 12 of 15 cases of
moderately differentiated OSCC, and 12 of 15 cases of moderately to poorly
differentiated OSCC, thus concluding that the presence of hCG-β positive tumor cells
appears potentially to reflect a malignant behavior of OSCC.
Hotakainen K et al (2003) studied expression of free hCG-β subunit in Renal Cell
Carcinoma (RCC) both in serum and tumor specimens. The prognostic value of hCG-β in
serum and tissue and its association with usual clinicopathological variables were
analysed. They concluded that serum concentration of hCG-β is an independent
prognostic variable in RCC and its tissue expression is not significantly associated with
prognosis.
6.3 Objectives of the Study:
1) To observe the level of hCG-β in serum of patients with oral squamous cell carcinoma
and normal volunteers.
2) To evaluate the degree of expression of hCG-β in tumor specimens of oral squamous
cell carcinoma immunohistochemically and to correlate with grades of oral squamous cell
carcinomas.
3) To correlate serum hCG-β levels and its expression in tumor specimens of different
grades of oral squamous cell carcinomas.
7 .Materials and Methods:
7.1 Source of Data:
Patients with oral squamous cell carcinoma reported to the Department of Head and
Neck Oncology, Mazumdar Shaw Medical Centre, Narayana Hrudayalaya, Bangalore.
7.2 Method of collection of data
7.2.1 Sample Size:

60 preoperative patients with oral squamous cell carcinoma.

20 volunteers.
7.2.2 Study Material:

Serum of patients with oral squamous cell carcinoma.

Serum of volunteers.

Formalin-fixed paraffin-embedded blocks of the tumor specimen of the same
patients.

ELISA kit.

IHC kit.
7.2.3 Study Method:
hCG-β levels will be evaluated in the serum of patients with oral squamous cell
carcinoma using ELISA kit. Tumor specimen from the same patients will be stained with
IHC kit using mouse anti-hCG-β antibody. In each case expression and quantification of
hCG-β will be performed in both serum and tumour specimens. Serum hCG-β levels of
patients will be compared with volunteers. The data collected will be subjected to
appropriate statistical analysis.
7.2.4 Study Design:
Cross sectional study
7.3 Inclusion Criteria:
1. Preoperative Oral Squamous Cell Carcinoma patients.
7.4 Exclusion Criteria:
1. Oral Squamous Cell Carcinoma patients undergoing treatment.
2. Oral Squamous Cell Carcinoma patients with history of treatment.
3. Patients with any other carcinomas.
7.5 Does the study require any investigation or interventions to be conducted on
patients or other humans or animals? If so, please describe briefly.
Yes.
5ml of blood sample and biopsy specimen will be collected from the patients with
oral squamous cell carcinoma and 5ml of blood sample will be collected from the
volunteers after obtaining informed consent from them.
7.6 Has ethical clearance been obtained from your institution for this study?
Yes.
The study would be conducted in the department of Head and Neck Oncology,
Mazumdar Shaw Medical Centre, Narayana Hrudayalaya, Bangalore and permission
has been obtained from the internal review board and the ethical committee of the
institute
8
List of References:
1) Hedstrom J, Grenman R, Ramsay H, Finne P, Lundin J, Haglund C et al.
Concentration of free hCG-β subunit in serum as a prognostic marker for
squamous cell carcinoma of the oral cavity and oropharynx. International Journal
of Cancer 1999 Oct 22; 84(5): p.525-528.
2) Kanokporn B, Abdel H.K, Dale A. Miles. Immunohistochemical study of the
expression of hCG-β in oral squamous cell carcinoma. American Cancer Society
1999 Feb15; 85 (4): p.757-762
3) IlesR K, Purkis P E, Whitehead P C, Oliver R T D, Leigh I , Chard T.
Expression of β-hCG by non-trophoblastic non-endocrine ‘normal’ and malignant
epithelial cells. Br. J. Cancer 1990; 61: p.663-6.
4) Sheaff M T, Martin J E, Badenoch D F, Baithun S I. β-hCG as a prognostic
marker in adenocarcinoma of the prostate. J. Clin Pathol 1996; 49: p.329-332
5) Marcillac I, Troalen F, Bidart J.. Free hCG-β subunit in gonadal and
nongonadal neoplasms. Cancer Res 1992; 52: p.3901-07
6) Hotakainen K, Ljungberg B, Haglund C, Nordling S, Paju A and Stenman
U.Expression of free β subunit of hCG in renal cell carcinoma: Prognostic study on
tissue and serum. Int J Cancer 2003; 104: p.631-635.
7) Alfthan H, Haglund C, Dabek J, and Sntenman U. Concentrations of Human
Choriogonadotropin, Its β-subunit, and the core fragment of the β-subunit in serum
and urine of men and nonpregnant women. Clinicalchemistry1992; 38:10.
8) Cole L, Kardana A, Ying F, Birken S. The biological and clinical significance of
nicks in Human Chorionic Gonadotropin and its free β- subunit. The Yale Journal
of Biology and Medicime 1991;64:627-637
9. Signature of the Candidate:
10. Remarks of the Guide:
11. Name and Designation of the Guide/s:
11.1.
Guide :
Dr. PUSHPALATHA MAHESH
PROFESSOR
Department of Oral and Maxillofacial Pathology,
AECS Maaruti College of Dental Sciences and Research
Centre,
Bangalore – 560 076
11.2.
Signature:
11.3.
Co-Guide (if any):
11.4.
Signature:
11.5.
Head of the Dept.:
DR. P.SHARADA,
PROFESSOR & HEAD,
Department of Oral and Maxillofacial Pathology,
AECS Maaruti College of Dental Sciences and Research
Centre,
Bangalore – 560 076
11.6. Signature:
12. Remarks of the Chairman and Principal:
12.1.
Signature:
PATIENT CONSENT FORM
Name:
Age :
Sex:
Address :
Biopsy obtained from (site):
I hereby give my consent to the Department of Head and Neck Oncology,
Mazumdar Shaw Medical Centre, Narayana Hrudayalaya, Bangalore to use my blood
sample and the biopsy specimen (Incisional/Excisional) for research purpose.
Signature of the patient
RAJIV GANDHI UNIVERSITY OF
HEALTH SCIENCES
BANGALORE, KARNATAKA
MASTER OF DENTAL SURGERY (MDS)
ORAL AND MAXILLOFACIAL PATHOLOGY
2013
A.E.C.S MAARUTI COLLEGE OF
DENTAL SCEINCES
& RESEARCH CENTRE, BANGALORE – 560 076