3. Illustration 11.5: Photomicrograph of histopathological section of well differentiated OSCC (Case no. 30;
H&E stain, 100X magnification).
4. Illustration 11.6: Photomicrograph of histopathological section of well differentiated OSCC (Case no. 30;
H&E stain, 400X magnification).
5. Illustration 11.7: Photomicrograph of a section showing in well differentiated OSCC , E-cadherin reduced
expression<44% ( 2)×(2)Intensity =4(Case no: 30; IHC, 400X magnification).
6. Illustration 11.8: Photomicrograph of a section showing E-cadherin in well differentiated OSCC reduced
expression>44% ( 2)×(2)Intensity =4(Case no: 30; IHC, 400X magnification).
7. Illustration 11.13: Photomicrograph of histopathological section of moderately differentiated
OSCC (Case no. 33; H&E stain, 400X magnification).
8. Illustration 11.14: Photomicrograph of histopathological section of moderately differentiated
OSCC (Case no. 33; H&E stain, 400X magnification).
9. Illustration 11.15: Photomicrograph of a section showing E-cadherin in moderately differentiated
OSCC reduced expression<09%% ( 1)×(1)Intensity =1(Case no: 33; IHC, 400X magnification).
10. Illustration 11.16: Photomicrograph of a section showing E-cadherin in moderately differentiated
OSCC reduced expression<09%% ( 1)×(1)Intensity =1(Case no: 33; IHC, 100X magnification).
11. Illustration 11.21: Photomicrograph of histopathological section of poorly differentiated OSCC
(Case no. 41; H&E stain, 100X magnification).
12. Illustration 11.22: Photomicrograph of histopathological section of poorly differentiated OSCC
(Case no. 41; H&E stain, 400X magnification).
13. Illustration 11.23: Photomicrograph of a section showing E-cadherin in poorly differentiated
OSCC reduced expression <47% ( 2)×(1)Intensity =2(Case no:41; IHC, 100X magnification).
14. Illustration 11.24: Photomicrograph of a section showing E-cadherin in poorly differentiated
OSCC reduced expression< 47%% ( 2)×(1)Intensity =2(Case no:41; IHC, 100X magnification).
15.
16. Introduction
Oral cancer is one of the topmost malignancies worldwide.
Oral cancer mostly affects in the area of lips, tongue, mouth,
salivary glands and oropharynx (Sultana & Malik 2014). Oral
squamous cell carcinoma comprises 90% of all Head-Neck
squamous cell carcinomas (Dhingra et al. 2017).
19. Introduction(contd.)
The use of tobacco and/or the areca nut (betel) yields
potentially malignant injuries from which oral cancer
develops particularly in developing countries. Consumption of
alcohol is also important in oral carcinogenesis particularly in
developed countries.
20. Introduction(contd.)
Other predisposing factors of OSCC include dietary habits
(decrease consumption of fresh vegetables and increase
consumption of meat), immunodeficiency, human papilloma
virus (HPV 16/18) infection and poor oral hygiene (Hossain et
al., 2015).
21. Introduction(contd.)
Various genetic events occur in oral carcinogenesis that leads to the
disruption of the normal regulatory pathways of cell cycle (Gatoo &
Dar, 2018).The ordered cells enter into G1, S, G2 and M phases of cell
cycle sequentially and the progression is driven by the proteins named
“cyclins,” which are activated by binding with specific cyclin-
dependent kinases (CDK) (Saawarn et al., 2012).
22. Introduction(contd.)
Cyclin D1 is one of the strongly implicated cyclins in human
carcinogenesis. The cyclin D1 gene (CCND1) is located on chromosome
11q13 and encodes a nuclear protein that binds with cyclin-dependent
kinases 4 and 6, which phosphorylate and inactivate the retinoblastoma
protein (pRb) thereby, allowing the progression of genetically damaged
cells from G1 to S phase of the cell cycle (Gioacchini et al., 2014).
23. Introduction(contd.)
In addition to this, G1 phase is shortened and dependency on
growth factors for cell proliferation is also reduced (Basnaker,
Srikala & Satish, 2014).
24. Introduction(contd)
An increased level of cyclin D1 expression is often closely
related with tumor size, nodal involvement, poor
differentiation, advanced stage and also non-response to
treatment and poor survival (Ramos-Garcia et al., 2017).
25. Introduction(contd.)
Cyclin D1 overexpression promotes genetically damaged cell
proliferation and also inhibits Cisplatin induced tumor cell
apoptosis (Zhou et al., 2009). That’s why, it may be an important
factor for future targeted therapy (Palbociclib-under trial) in
patients of OSCC (Hamilton and Infante, 2016).
26. Rationale of the study
Oral cancer is one of the most common malignancies in head neck
region and squamous cell carcinoma is the most common histologic
type of oral cancer. The spectrum of epithelial alterations that are
commonly seen in the oral lesion range from keratosis (leukoplakia,
erythroplakia), squamous hyperplasia, verrucous hyperplasia, dysplasia
(mild, moderate, severe), carcinoma in situ to invasive carcinoma
27. Rationale (contd.)
For diagnosis of oral cancer, histopathology remains gold
standard but several other ancillary techniques can help to
assess the biological behavior of the tumor.
28. Rationale (contd.)
Several studies have been showed that significant
immunohistochemical expression of cyclin D1 occurs in the
patients of OSCC. Overexpression of cyclin D1 represents
aggressiveness of the disease thereby indicating poor prognosis
and also indicates conventional chemo-resistance.
29. Rationale (contd.)
Therefore, the present study may be an important adjunct
along with grading, to determine the prognosis and also to
plan the treatment options that would lead to lesser morbidity
and increased survival of patients with OSCC.
33. Objectives (contd.)
Specific objectives
•To assess the histopathological grades of oral squamous cell
carcinoma.
•To assess the expression pattern (intensity & distribution) of the
cyclin D1 protein in different grades of oral squamous cell
carcinoma.
34. Objectives (contd.)
•To determine the association of cyclin D1 immunoexpression
with histopathological grades, age, gender, anatomic site and
the habit of using tobacco in OSCC
35. Materials and methods
•Study design: Cross sectional observational study.
•Place of study: Department of Pathology, Sylhet MAG Osmani
Medical College, Sylhet.
•Period of study: From March, 2019 to April, 2020 and July, 2021
to April, 2022.
36. Materials and methods (contd.)
•Study population: Tissue samples that diagnosed as oral
squamous cell carcinoma by histopathological examination.
•Sampling technique: Non-random convenience sampling.
•Sample size: n= 53.
40. IHC scoring system of Cyclin D1
Expression score
(ES)
% of cells stained
1 1-25
2 26-50
3 51-75
4 >75
Intensity score
(IS)
Intensity of staining
1 Mild
2 Moderate
3 Strong
Total score (TS=ESXIS) Grading
1-4 + (weak)
5-8 ++ (moderate)
9-12 +++ (strong)
41. Working flow chart
Target population
Histological diagnosis of OSCC (study population, n=52)
IHC of histopathological sections to see the cyclin D1 immunoexpression
Evaluation and scoring of cyclin D1 immunoexpression in OSCC
Data were recorded
Histopathological examination was done from oral lesions in routine H & E
stained sections
Statistical analysis was done
Writing of Thesis
Gradings of OSCC were done
43. Table 1: Distribution of the study cases according to age
(n=52)
Age groups (years) Frequency Percent
≤40 7 13.5
41-50 19 36.5
51-60 14 26.9
61-70 9 17.3
>70 3 5.8
Total 52 100.0
Mean ± SD 54.37 ± 12.92
Age range 30-90 years
44. Figure 1: Pie chart showing the frequency of the
study cases according to gender (n=52)
Male
28 cases
(53.8%) Female
24 cases
(46.2%)
45. Table 2: Distribution of the study cases according to
the habit of using tobacco and others (n=52)
Habits Frequency Percent
Habit of smoking tobacco 27 51.9
Habit of using smokeless tobacco 41 78.8
Habit of using both type tobacco 16 30.8
Habit of alcohol consumption 00 00
46. Figure 2: Bar diagram of the study cases according to
anatomic site of OSCC (n=52)
47. Figure 3: Pie chart showing the frequency of the study cases according to
histologic grade
48. Figure 4: Pie chart showing the frequency of study cases based on
the total score (TS) of cyclin D1 immunoexpression (n=52)
49. Table 3: Association between cyclin D1 immunoexpression and histologic grades
of OSCC
*Fisher Exact test was done to measure the level of significance.
Figure within parenthesis indicates corresponding percentage.
s idicates significant.
Histologic grades of OSCC Cyclin D1 immunoexpression p value
Weak positive Moderate positive Strong positive
Well differentiated 36 (87.8%) 4 (9.8%) 1 (2.4%)
Moderately differentiated 4 (50%) 1 (12.5%) 3 (37.5%) *0.014s
Poorly differentiated 2 (66.7%) 1 (33.3%) 0 (0.0%)
Total 42 (80.8%) 6 (11.5%) 4 (7.7%)
50. Table 4: Comparison of cyclin D1 immunoexpression between intergroup
histologic grades of OSCC
*Fisher Exact test was done to measure the level of significance.
Figure within parenthesis indicates corresponding percentage.
s idicates significant.
Intergroup histologic grade P value
Between well and moderately differentiated *0.008s
Between well and poorly differentiated *0.363
Between moderately and poorly differentiated *0.509
51. Table 5: Comparison of histologic grades of OSCC with patient’s age
Fisher Exact test was done to measure the level of significance.
Figure within parenthesis indicates corresponding percentage.
s idicates significant.
Age group (years) Histologic grades of OSCC p value*
Well differentiated Moderately
differentiated
Poorly differentiated
≤40 6 (85.7) 1 (14.3) 0 (0.0)
41-50 15 (78.9) 4 (21.1) 0 (0.0)
51-60 13 (92.9) 0 (0.0) 1 (7.1)
61-70 6 (66.7) 1 (11.1) 2 (22.2) P=0.047s
>70 1 (33.3) 2 (66.7) 0 (0.0)
Total 41 (78.8) 8 (15.4) 3 (5.8)
53. Conclusion
OSCC is the most common malignancy in the head-neck region.
The contribution of cyclin D1 expression in OSCC is significant
according to the present study. In most of the studies, CCND1
aberration has been proved as a prognostic marker for disease-free
survival and overall survival in OSCC patients.
54. Conclusion(contd.)
Deregulated cyclin D1 overexpression is associated with cisplatin based
conventional chemoresistance. So, it may be an important factor for
future targeted therapy (Palbociclib-under trial) in patients of OSCC.
Therefore, evaluation of cyclin D1 expression in pre-operative biopsy
specimen may be useful in predicting and planning for most appropriate
treatment strategies in patients with OSCC.
55. Limitations
•Mostly biopsy samples are studied. Resected samples will give
more informative results.
•Small sample size
•Out of 52 samples, only three cases of poorly differentiated
OSCC were available.
•Data for alcohol consumption was not found.
56. Recommendations
•Large sample size and resected specimens would bring more
representative data.
•A further study is needed to see the association of cyclin D1
immunoexpression with pathologic staging.
•Large cohort studies will give more precise results regarding
targeted therapy and overall survival.
•Molecular methods of cyclin D1 gene detection might be used
for more accurate results.
69. Illustration(contd.)
Photomicrograph of histopathological
section of moderately differentiated OSCC
(Case no. 6; H&E stain, 100X
magnification).
Photomicrograph of histopathological
section of moderately differentiated OSCC
(Case no. 6; H&E stain, 400X
magnification).
70. Illustration(contd.)
Photomicrograph of histopathological
section of poorly differentiated OSCC (Case
no. 53; H&E stain, 100X magnification).
Photomicrograph of histopathological
section of poorly differentiated OSCC (Case
no. 53; H&E stain, 400X magnification).
71. Illustration(contd.)
Photomicrograph of a section of tonsillar tissue taken as control showing cyclin
D1 expression confined to the basal layers of the epithelium (IHC, 100X
magnification).
75. Illustration(contd.)
Photomicrograph of a section showing
cyclin D1 positivity in <25% of the tumor
cells (Case no: 25, ES: 1; IHC, 400X
magnification).
Photomicrograph of a section showing
cyclin D1 positivity in 25-50% of the tumor
cells (Case no: 15, ES: 2; IHC, 400X
magnification).
76. Illustration(contd.)
Photomicrograph of a section showing
cyclin D1 positivity in 50-75% of the tumor
cells (Case no: 31, ES: 3; IHC, 400X
magnification).
Photomicrograph of a section showing
cyclin D1 positivity in >75% of the tumor
cells (Case no: 21, ES: 4; IHC, 400X
magnification).
