An Automated Approach to Analyze MMP1 Expression in Oral Squamous Cell Carcinoma Using Fluorescence in Situ Hybridization: An Image Based Analysis

Madhura, Murittige Gopalakrishna; Vijayalakshmi, Kotrashetti S; Veerendra, Kumar B; Sunil, Vasudev; Suchetha, Aghanashini; Kiran, Kumar; Preethi, Patil N; Rakshith, R; Ranjith, Kumar J; Manohar, Kugaji S; Kishore, Bhat S; Naveen, Krishnamoorthy; Sidramesh, Muttagi S; Patil, BR; Gopinath, Kodaganur Srinivaschar; Ramakant, Nayak S

doi:10.31557/APJCP.2025.26.7.2559

An Automated Approach to Analyze MMP1 Expression in Oral Squamous Cell Carcinoma Using Fluorescence in Situ Hybridization: An Image Based Analysis

Document Type : Research Articles

Authors

Murittige Gopalakrishna Madhura ¹^{, 2}^{, 3}
Kotrashetti S Vijayalakshmi ³^{, 4}
Kumar B Veerendra ³^{, 5}
Vasudev Sunil ³^{, 6}
Aghanashini Suchetha ³^{, 7}
Kumar Kiran ⁸
Patil N Preethi ⁹^{, 10}
R Rakshith ⁹^{, 10}
Kumar J Ranjith ⁹^{, 10}
Kugaji S Manohar ¹¹
Bhat S Kishore ¹²
Krishnamoorthy Naveen ¹³
Muttagi S Sidramesh ¹⁴
BR Patil ¹⁵
Kodaganur Srinivaschar Gopinath ¹⁶
Nayak S Ramakant ¹⁷

¹ Department of Oral Pathology and Microbiology, Maratha Mandal’s NGH Institute of Dental Sciences and Research Centre, Belagavi, India.

² Department of Oral Pathology and Microbiology, DAPM R V Dental College, Bengaluru, Karnataka, India.

³ Rajiv Gandhi University of Health Sciences Karnataka, India.

⁴ Department of Oral & Maxillofacial Pathology and Microbiology Maratha Mandal’s Nathajirao G Halgekar Institute of Dental Sciences and Research Centre, Belagavi, India.

⁵ Department of Oral Pathology and Microbiology, DAPM R V Dental College Bengaluru, India.

⁶ Department of Oral and Maxillofacial Surgery, DAPM R V Dental College, Bengaluru, India.

⁷ Department of Periodontics DAPM R V Dental College Bengaluru, India.

⁸ Department of Oral and Maxillofacial Pathology, SDM College of Dental Sciences and Hospital, A Constituent Unit of Shri Dharmasthala Manjunatheshwara University, Dharwad, Karnataka, India.

⁹ Department of Master of Computer Applications RV College of Engineering Mysore Rd, RV Vidyaniketan Post Bengaluru, India.

¹⁰ Visvesvaraya Technological University, Belagavi Karnataka, India.

¹¹ Aassistant Director Central Research Laboratory Maratha Mandal’s Nathajirao G Halgekar Institute of Dental Sciences and Research Centre 47 A/2, Bauxite Road, near KSRP Ground Belagavi - 590010 Rajiv Gandhi University of Health Sciences, Karnataka India.

¹² Former Director Central Research Laboratory Maratha Mandal’s Nathajirao G Halgekar Institute of Dental Sciences and Research Centre and Consultant, Arihant Hospital Belagavi - 590010 Karnataka, India.

¹³ Consultant General Pathologist Koushik Laboratory and Clinic Bengaluru - 560061 Karnataka, India.

¹⁴ Department of Oral and Maxillofacial Surgery KLE VK Institute of Dental Sciences VGJ8+JM9, Nehru Nagar Belagavi - 590010 KLE University Karnataka, India.

¹⁵ Chief Surgical Oncologist Karnataka Cancer Therapy and Research Institute Hubballi - 580025 Karnataka, India.

¹⁶ Consultant Surgical Oncologist HCG Cancer Centre And Ambuja Health Care Clinic No.2/2, Armugam Circle, Pattalamma Temple Rd, Basavanagudi Bengaluru, 560004 Karnataka, India.

¹⁷ Principal, Maratha Mandal’s NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, Rajiv Gandhi University of Health Sciences, Karnataka, India.

10.31557/APJCP.2025.26.7.2559

Abstract

Objectives: 1.To evaluate matrix metalloproteinase 1 (MMP1) expression in oral squamous cell carcinoma (OSCC) through fluorescence in situ hybridization (FISH). 2. To design an algorithm for the validation of FISH photomicrographs using a computer programming language. Material And Methods: The formalin fixed paraffin embedded (FFPE) tissue blocks from 20 histopathologically confirmed cases of OSCC and 24 normal oral mucosal (NOM) samples from healthy volunteers were considered. The inclusion and exclusion criteria were defined. After identifying the probe for MMP1, the protocol was optimized. 4 µm tissue sections were subjected to FISH for MMP1 analysis. The expression was observed manually under fluorescence microscope by two pathologists independently. With computer programme, an algorithm was designed to quantify the data from the microscopic images of FISH-MMP1. Results: The MMP1 expression was significantly more in OSCC when compared to NOM. The mean scores for staining intensity were significantly higher in OSCC [2.10 ± 0.72] as compared to NOM [1.54 ± 0.66] with p=0.01 and 95%CI [0.14, 0.98]. The ROC curve analysis showed a sensitivity of 80%, specificity of 54.2% and an accuracy of 65.9%. Further, the digital data quantification revealed the precision of the model used, to be 66.67 and recall 82.36, suggesting differential expression of MMP1 between OSCC and normal oral samples. Conclusion: The present study has shown greater expression of MMP1 in OSCC when compared to normal oral mucosal tissues. Further, a simple yet novel digital method to objectively quantify the expression of MMP1 in OSCC has been shown.

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