Comparison of Morphometric Assessment Using Methyl Green Pyronin and AgNOR Staining of Oral Squamous Cell Carcinoma
NCT ID: NCT01987934
Last Updated: 2013-11-20
Study Results
The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.
Basic Information
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UNKNOWN
60 participants
OBSERVATIONAL
2013-06-30
2013-12-31
Brief Summary
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Nuclear organizer regions (AgNORs) are associated with proliferative activity and represents as a diagnostic aid in oral malignancies. Similarly, methyl green pyronin (MGP) stain has also been valuable as a complement in routine histopathological studies of several neoplastic entities.
Morphometric techniques offer an opportunity to quantify nuclear changes associated with malignancy and may provide an objective basis for grading the tumors. The present study is planned to analyze the morphometric parameters of the MGP stain in oral squamous cell carcinoma, and in their various histological grades, and to assess if the MGP staining parameters could give information on the aggressiveness of the malignant lesions of oral cavity.
Sections from thirty cases of squamous cell carcinoma along with thirty cases of normal oral mucosa will be evaluated for methyl green pyronin (MGP) and AgNOR staining. Morphometric analysis of various MGP staining and AgNOR parameters would be performed using micrometer. Statistical analysis of the results will be carried out using SPSS. Quantitative variables will be expressed as mean ± Standard Deviation. Frequencies and percentages will be given for qualitative variables.
It is hypothesized that oral squamous cell carcinoma will exhibit significantly higher MGP staining and AgNOR staining parameters than normal mucosa of the oral cavity.
Detailed Description
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Out of the developing counties, Indo-Pak subcontinent amounts to one third of the global oral cancer burden. Carcinoma of the oral cavity in Pakistan, one of the high-risk countries is the second most frequent cancer in either gender. With differential distribution among sub-populations, it represents 8.8% of all reported cancer cases. The most predominant risk factors include tobacco smoking, betel chewing and alcohol.
Screening and specific diagnostic tools are two innovative approaches for early detection of oral cancer. Conventional oral examination and clinical evaluation remains the foundation of screening approaches whereas surgical biopsy followed by histological analysis is attributed to be the gold standard for diagnosing malignant oral lesions. Furthermore, some adjunctive techniques may contribute to the diagnosis of oral cancer. Of these vital staining, DNA analysis, chemiluminescence, tissue fluorescence, light-based detection systems and biomarkers are now on the forefront.
Biomarkers have been valuable in revealing additional information about tumor pathology, including the inactivation of tumor suppressor genes, angiogenesis, apoptosis and cell proliferation. Aberrations in proliferation kinetics are the most imperative factor during tumor progression. Three main categories of cell proliferation markers include growth fraction markers like Ki67, cycle-specific markers like PCNA(Proliferating Cellular Nuclear Antigen), and cell cycle time markers like AgNOR. During malignancy, proliferation and protein synthesis is mainly controlled by the nucleus.
Nucleolar Organizer Regions (NORs) are loops of DNA on the short arms of acrocentric chromosomes that presumably are associated with cell proliferation. Some of the NOR associated proteins are argyrophilic and can be demonstrated as black dots by the silver staining technique and the structures thus demonstrated are known as AgNORs. The AgNOR quantitative parameters provide information about the velocity of cell proliferation during cell cycle. Neoplastic progression is affected more by the rate of cell proliferation, rather than the proliferative activity per se.
Although AgNOR staining and AgNOR quantification are valuable methods in histopathological evaluation of malignancies, certain limitations have also been associated with this technique. Such AgNOR associated shortcomings include nucleolar affinity for silver stain obscuring the individual AgNORs in cases of intense staining, overlapping of variable degree between high and low grade tumors, difficulty in discerning doublets or clusters by light microscopy, misinterpreted stain deposition as proliferative activity along with some technical problems of staining and fixation.
Methyl Green Pyronin (MGP) stain is a differential stain used for selective staining of DNA and RNA. Methyl green binds specifically to deoxyribonucleic acid (DNA), staining nuclei green, while pyronin is specific for ribonucleic acid (RNA), staining nucleoli red. Methyl green pyronin stain has also been used to distinguish between proliferating and dedifferentiated non-proliferating cell nuclei. Similarly to assess the degradation of DNA in thyroid follicular epithelial cells, this stain has also proven to be useful.
Morphometry has been a significant advancement in delineating sequence of events responsible for cancer progression. Variations in nuclear and cellular size and shape have greater implications in the assessment of neoplastic lesions. Methyl Green Pyronin (MGP) stain has been attracting attention in the recent times as it has been used to evaluate various nuclear morphometric parameters in breast neoplasms. By using methyl green-pyronin staining, not only the inadequacies associated with AgNOR staining can be lessened but also the morphometric parameters for evaluating changes in the nucleus and nucleolus during carcinogenesis can be accomplished.
So far AgNOR staining appears to be a useful diagnostic tool in oral squamous cell carcinoma for examination of nucleolar structure and variations in nucleolar activity. On the other hand, MGP staining has been used in assessing the cancerous lesions of breast and thyroid gland and recently has been extended to oral lesions. However, no studies have been undertaken yet to evaluate and compare the morphometric parameters using these two stains within normal oral epithelium and oral squamous cell carcinoma. Similarly, very few studies have been undertaken regarding application of methyl green pyronin stain in oral pathologies.
Hence the current study has been planned to evaluate the morphometric parameters among normal oral epithelium and oral squamous cell carcinoma using methyl green pyronin staining; moreover these findings will also be compared with those of AgNOR staining. In this current scenario, methyl green pyronin staining and morphometric analysis may go hand in hand to provide a better diagnosis and early detection of oral cancer.
Conditions
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Keywords
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Study Design
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CASE_CONTROL
PROSPECTIVE
Study Groups
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Normal or Control group
Those subjects with normal oral epithelium will be included in this group.
No interventions assigned to this group
Study Group
Those subjects with oral squamous cell carcinoma will be included in this group.
No interventions assigned to this group
Eligibility Criteria
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Inclusion Criteria
* Cases of clinically and histologically confirmed primary oral squamous cell carcinoma based on WHO criteria
Exclusion Criteria
* Subjects having metastatic tumors and tumors other than in the oral cavity
* Subjects having recurrent tumors.
* Subjects of normal or control group with history of risk / predisposing factors for oral cancer.
ALL
Yes
Sponsors
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Postgraduate Medical Institute, Lahore
OTHER
Academy of Interdisciplinary Dental Education and Research
OTHER
Responsible Party
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Ali Raza
Dr. Ali Raza
Principal Investigators
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Muhammad Eyyaz Khaleel, M.Phil
Role: STUDY_DIRECTOR
Postgraduate Medical Institute, Lahore
Ali Raza, B.D.S
Role: PRINCIPAL_INVESTIGATOR
Postgraduate Medical Institute, Lahore
Locations
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Post Graduate Medical Institute
Lahore, Punjab Province, Pakistan
Countries
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References
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Mohtasham N, Mahdavi-Shahri N, Salehinejad J, Ejtehadi H, Torabi-Parizi M, Ghazi N. Detection of nucleoproteins in squamous cell carcinoma, and dysplastic and normal mucosa in the oral cavity by methyl green-pyronin staining. J Oral Sci. 2010 Jun;52(2):239-43. doi: 10.2334/josnusd.52.239.
Other Identifiers
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AIDER-1301-AR
Identifier Type: -
Identifier Source: org_study_id