 Abstract | | |
Aim: High-grade glial tumors remain as one of the most lethal malignancies. Cyclin D1 is expressed in some human malignancies and is the potential target of intervention. The present study aims to determine the relationship of cyclin D1 expression with other clinicopathological parameters. Materials and Methods: A cross-sectional study was carried out in a tertiary care center. Biopsy proven 66 cases of glial tumor patients were included in the study. The patients with incomplete clinical details were excluded from the study. Immunohistochemistry using antibodies for IDH 1 and cyclin d1 was done in all the cases. Glial tumors were reclassified according to WHO 2016 classification. Data analysis was performed using SPSS 26.0 for the windows. Result: Among 66 patients, 49 (74.3%) were males and 17 (25.7%) were females. The age of the patients ranged from 20 years to 70 years. Overall, 6.02% were of grade I Glial tumors, 22.7% were of grade II Glial tumors, 19.6% patients were of grade III Glial tumors, and 51.6% patients were of grade IV Glial tumors. Of 66 samples tested cyclin D1 was positive in 25 (37.87%) as high expressers and 7 (10.60%) were low expressers. Our study showed a significant correlation between the expression of cyclin D1 with grade and IDH mutation status, No significant correlation of cyclin D1 was noted with age or sex of the patient. Conclusion: Cyclin D1 was associated with a higher grade of the glial tumor. It can be a potential marker both for prognosis and treatment of glial tumors.
Keywords: Anaplastic astrocytoma, cyclin D1, diffusely infiltrating glioma, glioblastoma multiforme, IDH1
How to cite this article:
Mehta R, Yadav R, Vatsgotra R. Role of cyclin D1 in glial tumors—A retrospective and observational study. Indian J Pathol Microbiol 2023;66:264-8 |
| Introduction | |  |
Astrocytic tumors are the commonest tumors affecting the brain in all age groups. High-grade glial tumors remain as one of the most lethal malignancies with a median survival of 18 months. Despite studying various molecular alterations in glial tumors, its median survival remains low.
Glioblastoma multiforme is the most aggressive glial tumor, comprising 15.6% of all primary brain tumors.[1] It is a rare but highly aggressive brain tumor affecting about 19.3 patients per 1 l ac people.[2] GBM can affect an individual of any age, but it is commonly seen in the 4th to 7th decade.[3] GBM can arise de novo or can progress from low-grade glial tumor.[4]
Complete surgical resection is the mainstay of the treatment followed by radiotherapy and chemotherapy. Development in the treatment can increase the life expectancy in patients with GBM.[5] Thus, research that can lead to the development of novel treatment modalities is of extreme importance. Cyclin-dependent kinase inhibitors have shown a promising role in various malignancies. The cell cycle is regulated by a cyclin, cyclin-dependent kinases, and cyclin-dependent kinase inhibitor. Cyclin attaches itself to cyclin-dependent kinases and allows for further progression of the cell cycle. Cyclin D1-3 are involved in the transition from G1 to the S phase in the cell cycle.[6] Cyclin E activates CDK 4-6 and CDK 2.[7] Cyclin D1 overexpression is associated with a higher grade of the tumor. It increases the activity of metalloproteinases, thus enhancing the cell movement. Expression of Cyclin D1 has been studied in various tumors like breast, papillary carcinoma thyroid, and renal cell carcinoma.[8],[9],[10],[11] Expression of cyclin D1 is yet to be studied in glial tumors. Few researchers have studied the role of cyclin D1 in brain tumors. However, there is disagreement in the expression of cyclin D1 in low- and high-grade glial tumors. Hereby, the present study was conducted to find the role of cyclin d1 in glial tumors and its correlation with the grade of the tumor and IDH1 mutation.
[TAG:2]Materials and Methods[/TAG:2]
A cross-sectional study was carried out in a tertiary care center. Biopsy proved 66 cases of glial tumor patients from Dec 2015 to Dec 2019 were included in the study. The clinical details were retrieved from biopsy requisition forms. The patients with incomplete clinical details were excluded from the study. Immunohistochemistry using antibodies for IDH 1 (IDH1-R132H antibody clone H09) and cyclin D1 was done in all cases. H and E sections were reviewed. Based on IHC and histopathological findings, glial tumors were reclassified according to WHO 2016 classification. For immunohistochemistry 3-5 μ, thick sections were dewaxed and rehydrated, and epitope retrieval was done in a pressure cooker at 15 psi and 120°C for 10 min by incubating slides in citrate buffer (pH 6.0). Hydrogen peroxidase was used for blocking endogenous peroxidase activity. Thereafter, primary antibody was added and incubated for 1 hour and subsequently treated with biotinylated secondary antibody for another 30 minutes. This step was followed by treating the slide with Streptavidin horseradish peroxidase for 10 minutes. Diaminobenzidine (DAB) was used as the chromogen and counterstaining with hematoxylin was carried out. Subsequent IDH 1 and 2 gene mutation analysis by PCR-sequencing [tested for IDH1—R132H, R132C, R132S, R132G, R132L, R132V, R140W, R140L, and IDH2—R140G, R140Q, R172K, R172M, R172W, R172G] was done.
Glial tumors were reclassified according to WHO 2016 classification [Figure 1] and [Figure 2]. Diffuse astrocytic and oligodendroglial tumors included grade II, III, and grade IV glial tumors. These tumors were further subclassified based on histology and molecular markers. WHO grade was determined based on histology. Diffuse astrocytic tumors included diffuse astrocytoma IDH—mutant [Figure 3], diffuse astrocytoma IDH wild type [Figure 4], gemistocytic astrocytoma–IDH mutant. Grade III glial tumors included anaplastic astrocytoma IDH—wild type, anaplastic astrocytoma IDH mutant. WHO grade IV included glioblastoma IDH mutant and glioblastoma IDH wild type. Other astrocytic tumors included grade I glial tumors, namely, pilocytic astrocytoma. One case of Mixed oligoastrocytoma was noted. ATRX and TP 53 and deletion 1p/19q were done in this case. Based on ATRX loss, TP53 mutation, and IDH mutant, mixed oligoastrocytoma was reclassified as diffuse astrocytoma IDH mutant.
Immunohistochemistry using Cyclin D1 antibody was done in all cases. Areas with a high density of tumor cells were selected. The percentages of cyclin D1 immunopositive tumor cells were counted in 5 consecutive microscopic fields (magnification 100x). One hundred tumor cell nuclei were evaluated per field and the mean for each of the 5 fields was calculated. A granular nuclear stain was scored as positive. Cyclin D was considered positive when at least 5% of tumor cells showed nuclear positivity. Tumors with less than 50% cyclin D1-positive tumor cells were considered the low expression, whereas those with >50% cyclin D1-positive tumor cells were considered high expression [Figure 5]. Data analysis was performed using SPSS 26.0 for the Windows student version. Approval from institutional ethics commitee was obtained for the study on 21/12/2020.
| Results | | |
Total 66 cases were included in the present study. Among 66 patients, 49 (74.3%) were males and 17 (25.7%) were females. Males to female ratio was 2.9:1. The age of the patients ranged from 20 years to 70 years. Maximum patients fell in the age group between 40 and 49 years.
Based on IHC, glial tumors were divided into IDH 1 mutated or IDH wild type. IDH mutant type was noted in 24 (36.3%) patients and IDH1 wild type was noted in 42 (63.3%) of patients [Table 1].
Of 66 samples tested cyclin D1 was positive in 25 (37.31%) as high expressers and 7 (10.44%) were low expressers. The present study demonstrated that out of 14 diffuse infiltrating astrocytomas, IDH 1 wild type was seen in 6 (42.85%) patients and IDH 1 mutant was seen in 8 (57.14%) patients [Table 2]. Among 13 cases of anaplastic astrocytoma, IDH mutant type was seen in 8 (61.5%) and IDH wild type was seen in 5 (38.4%) patients [Table 3]. Among 34 patients of GBM, IDH 1 wild type was seen in 26 (76.4%) patients and IDH mutant type was seen in 8 (23.5%). Eight cases of secondary GBM were included in the study. Seven cases of secondary GMB were IDH 1 mutant type. Our study showed a significant correlation between expression of cyclin D1 expression with the WHO grade and IDH wild type, Cyclin D1 was strongly correlated with the WHO grade of the tumors, and the expression of the cyclin D1 increased as the grade of the glial tumor grade increased and 24 (70.58%) of WHO grade IV tumors were high expressers of cyclin D1. Patients with wild-type IDH overexpressed Cyclin D1. No correlation was found between age and sex.
| Discussion | | |
Prior classification of brain tumors was based predominantly on the cell of origin, histopathological findings, and grade of differentiation. In 2016, WHO revised the classification of brain tumors and molecular parameters were incorporated along with pre-existing histological features. Phenotypic and genotypic parameters were included in the study which led to greater diagnostic accuracy and improved patient management. In our study, we reclassified glial tumors according to WHO 2016 classification and correlated the grade of glial tumors with cyclin D1 expression. According to the 2016 classification, diffusely infiltrating gliomas including both astrocytic and oligodendroglial tumors is grouped, based on their growth pattern and IDH1 and IDH 2 gene mutations. The diffuse gliomas include the WHO grade II and grade III astrocytic tumors, the grade II and III oligodendrogliomas, and the grade IV glioblastomas. Astrocytoma with circumscribed growth pattern, lacking IDH gene family alterations, or TSC1/TSC2 mutations was excluded from the diffuse astrocytoma category and was classified as separate entities. This group included pilocytic astrocytoma and subependymal giant cell astrocytoma. The WHO grade II diffuse astrocytoma's and WHO grade III anaplastic astrocytoma are each divided into IDH-mutant, IDH-wildtype, and NOS categories based on the presence of IDH 1 and IDH2 positivity. Glioblastomas are subclassified as glioblastoma IDH-wildtype, which occurs primarily in primary or de novo glioblastoma, glioblastoma IDH-mutant which occurs as secondary glioblastoma, and glioblastoma NOS, in which full IDH studies are not done. Cyclin D1 plays a crucial role in cell cycle.[12] Studies performed previously showed that overexpression of Cyclin D1 is associated with a high grade of the tumor. The role of cyclin D1 has been established in breast malignancies, colorectal cancer, prostate cancer, and lymphomas. In the present study, 1.5% of patients of diffuse astrocytoma were cyclin D1 positive with less than 50% cells showing granular nuclear positivity. Three percent of patients of anaplastic astrocytoma showed low expression of cyclin D1, whereas 6% of anaplastic astrocytoma showed high expression of cyclin D1 positivity. 36.3% of cases of GBM showed high expression of cyclin D1 and 3% cases of GBM showed cyclin D1 positivity with low expression of cyclin D1. Our results are in accordance with other studies done in past. A study conducted by Parvin Mahzouni et al.[13] in 2019 demonstrated that cyclin D1 was positive in 60% of high-grade gliomas. A study conducted in 2007 showed that overexpression of cyclin D1 was noted in 40%, 12%, 52%, and 58% in breast cancer, colon tumors, glioblastomas, and melanomas.[14] In a similar study conducted by Qu Dw et al.[15] on gliomas in 2014, the expression of cyclin D1 in gliomas was 59%. In the present study, cyclin D1 expression was different in different grades of the tumor and it increased as the grade of the tumor increased. Tan et al.[16] in 2004 reported higher expression of cyclin D1 in high-grade glioma. According to their study, 31.25% and 61.53% of low-grade and high-grade tumors showed cyclin D1 positivity, respectively. Zhang et al.[17] studied 52 samples of gliomas and they reported that cyclin D1 is closely related to tumor formation as well as tumor progression. A study done by A Chakrabarty et al.[18] on gliomas also demonstrated higher expression of cyclin D1 in high-grade gliomas. In total, 8 secondary GBM were included in this study, cyclin D1 was positive in one case with more than 50% cells showing positivity, whereas other 7 secondary gliomas were IDH 1 mutant type. No significant association of cyclin D1 was noted with the age and sex of the patient. This is per the study conducted by Zhang YY et al. wherein they studied the association of cyclin D1 with age and sex of patients with advanced squamous cell carcinoma larynx.[19] This study concluded that with increasing grade of glial tumors, expression of cyclin D1 increases and expression of cyclin D1 is associated with IDH1 wild type.
Limitation of the study
The number of secondary GBM included in the present was very less. Hence, for validating the role of cyclin D1 in secondary glioma, more studies with the increased number of secondary GBM are required.
| Conclusion | | |
Cyclin D1 is associated with tumor progression and hence can be used as one of the prognostic markers of glioma and can be a potential target for treatment.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
| References | | |
| 1. | Morana G, Tortora D, Stagliano S, Nozza P, Mascelli S, Severino M, et al. Pediatric astrocytic tumor grading: comparison between arterial spin labeling and dynamic susceptibility contrast MRI perfusion. Neuroradiology 2018;60:437–46.  |
| 2. | Ostrom QT, Gittleman H, Farah P, Ondracek A, Chen Y, Wolinsky Y, et al. CBTRUS statistical report: Primary brain and central nervous system tumors diagnosed in the United States in 2006-2010. Neuro Oncol 2013;15(Suppl 2):ii1-56. |
| 3. | Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 2015;136:E359–86. |
| 4. | Cai X, Sughrue ME. Glioblastoma: New therapeutic strategies to address cellular and genomic complexity. Oncotarget 2018;9:40–54. |
| 5. | Gately L, McLachlan SA, Dowling A, Philip J. Life beyond a diagnosis of glioblastoma: A systematic review of the literature. J Cancer Surviv 2017;11:447–52. |
| 6. | Koontongkaew S. The tumor microenvironment contribution to development, growth, invasion and metastasis of head and neck squamous cell carcinomas. J Cancer 2013;4:66–83. |
| 7. | Liu S, Yin F, Fan W, Wang S, Guo XR, Zhang JN, et al. Over-expression of BMPR-IB reduces the malignancy of glioblastoma cells by upregulation of p21 and p27Kip1. J Exp Clin Cancer Res 2012;31:52. |
| 8. | Lee SH, Lee JK, Jin SM, Lee KC, Sohn JH, Chae SW, et al. Expression of cell-cycle regulators (cyclin D1, cyclin E, p27kip1, p57kip2) in papillary thyroid carcinoma. Otolaryngol Head Neck Surg 2010;142:332–7. |
| 9. | Aaltonen K, Amini RM, Landberg G, Eerola H, Aittomaki K, Heikkila P, et al. Cyclin D1 expression is associated with poor prognostic features in estrogen receptor positive breast cancer. Breast Cancer Res Treat 2009;113:75–82. |
| 10. | Lima MS, Pereira RA, Costa RS, Tucci S, Dantas M, Muglia VF, et al. The prognostic value of cyclin D1 in renal cell carcinoma. Int Urol Nephrol 2014;46:905–13. |
| 11. | Pereira RA, Ravinal RC, Costa RS, Lima MS, Tucci S, Muglia VF, et al. Cyclin D1 expression in prostate carcinoma. Braz J Med Biol Res 2014;47:515–21. |
| 12. | Wu WS, Chien CC, Liu KH, Chen YC, Chiu WT. Evodiamine prevents glioma growth, induces glioblastoma cell apoptosis and cell cycle arrest through JNK activation. Am J Chin Med 2017;45:879–99. |
| 13. | Mahzouni P, Taheri F. An immunohistochemical study of cyclin d1 expression in astrocytic tumors and its correlation with tumor grade. Iran J Pathol 2019;14:252-7. |
| 14. | Kleiner HE, Krishnan P, Tubbs J, Smith M, Meschonat C, Shi R, et al. Tissue microarray analysis of eIF4E and its downstream effector proteins in human breast cancer. J Exp Clin Cancer Res 2009;28:5. |
| 15. | Qu DW, Xu HS, Han XJ, Wang YL. Expression of cyclinD1 and Ki-67 proteins in gliomas and its clinical significance. Eur Rev Med Pharmacol Sci 2014;18:516–9. |
| 16. | Tan PG, Xing Z, Li ZQ. Expression of cyclin D1 in brain gliomas and its significance. Ai Zheng 2004;23:63–5. |
| 17. | Zhang X, Zhao M, Huang AY, Fei Z, Zhang W, Wang XL. The effect of cyclin D expression on cell proliferation in human gliomas. J Clin Neurosci 2005;12:166–8. |
| 18. | Chakrabarty A, Bridges LR, Gray S. Cyclin D1 in astrocytic tumours: An immunohistochemical study. Neuropathol Appl Neurobiol 1996;22:311–6. |
| 19. | Zheng Y, Shen H, Sturgis EM, Wang LE. Cyclin D1 polymorphism and risk for squamous cell carcinoma of the head and neck: a case–control study. Carcinogenesis. 2001;22:1195-9. |
Correspondence Address:
Rashmi Yadav
Department of Pathology INHS Asvini, Mumbai
India
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/IJPM.IJPM_1439_20
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3] |
