Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation

Abstract Background Thyroid cancer, characterized by significant genetic and epigenetic alterations, remains a critical focus of molecular oncology. This study investigates eight key genes (BRAF, EIF1 AX, FOXE1, KRAS, PDGFRA, PIK3 CA, PTEN, and TERT) that are deregulated in Hashimoto's Thyroidi...

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Main Authors: Mostafa A. Abdel-Maksoud, Taghreed N. Almana, Saeedah Almutair, Abdulaziz Alamri, Ibrahim A. Saleh, Mohamed Y. Zaky, Wahidah H. Al-Qahtani, Yasir Hameed
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Language:English
Published: BMC 2025-05-01
Series:Hereditas
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Online Access:https://doi.org/10.1186/s41065-025-00429-0
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author Mostafa A. Abdel-Maksoud
Taghreed N. Almana
Saeedah Almutair
Abdulaziz Alamri
Ibrahim A. Saleh
Mohamed Y. Zaky
Wahidah H. Al-Qahtani
Yasir Hameed
author_facet Mostafa A. Abdel-Maksoud
Taghreed N. Almana
Saeedah Almutair
Abdulaziz Alamri
Ibrahim A. Saleh
Mohamed Y. Zaky
Wahidah H. Al-Qahtani
Yasir Hameed
author_sort Mostafa A. Abdel-Maksoud
collection DOAJ
description Abstract Background Thyroid cancer, characterized by significant genetic and epigenetic alterations, remains a critical focus of molecular oncology. This study investigates eight key genes (BRAF, EIF1 AX, FOXE1, KRAS, PDGFRA, PIK3 CA, PTEN, and TERT) that are deregulated in Hashimoto's Thyroiditis and their roles in thyroid cancer. Methods Cell culture, nucleic acid extraction, RT-qPCR, bisulfite sequencing, and various in silico tools and databases. Results Expression analysis using RT-qPCR revealed significant (p-value < 0.05) down-regulation of BRAF, EIF1 AX, FOXE1, KRAS, PDGFRA, PIK3 CA, PTEN, and TERT genes in thyroid cancer cell lines compared to controls, with ROC curves indicating high diagnostic accuracy (AUC 0.93–0.99). Bisulfite sequencing demonstrated increased promoter methylation across all eight genes in cancerous samples, suggesting epigenetic silencing as a regulatory mechanism. Validation through UALCAN, OncoDB, and HPA confirmed reduced gene and protein expression in additional thyroid cancer cohorts. Genetic alteration analysis via cBioPortal showed prevalent BRAF mutations, whereas other genes exhibited fewer alterations. Kaplan–Meier survival analysis linked lower expression of BRAF and PIK3 CA to poorer overall survival. Correlation studies using TISIDB and TISCH2 databases highlighted associations between gene expression and immune modulation, revealing significant correlations with immune cell infiltration and diverse immune subtypes. Moreover, miRNA-mRNA network analysis identified hsa-mir- 628 - 5p as a critical regulator targeting these genes. The impact of BRAF overexpression on SW579 cells was assessed through various functional assays. Overexpression of BRAF resulted in reduced cell proliferation, colony formation, and wound healing, which may reflect context-dependent effects. While BRAF is typically oncogenic, its overexpression may lead to cellular stress or negative feedback mechanisms that impair these processes. Conclusion This comprehensive analysis elucidates the complex regulatory landscape of these genes in thyroid cancer, emphasizing the significant role of epigenetic modifications and providing insights into potential diagnostic and therapeutic avenues.
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spelling doaj-art-c8e01e83a4034ba7aa0c20e9c6cb14292025-08-20T01:59:57ZengBMCHereditas1601-52232025-05-01162112010.1186/s41065-025-00429-0Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validationMostafa A. Abdel-Maksoud0Taghreed N. Almana1Saeedah Almutair2Abdulaziz Alamri3Ibrahim A. Saleh4Mohamed Y. Zaky5Wahidah H. Al-Qahtani6Yasir Hameed7Department of Botany and Microbiology, College of Science, King Saud UniversityDepartment of Botany and Microbiology, College of Science, King Saud UniversityDepartment of Botany and Microbiology, College of Science, King Saud UniversityBiochemistry Department-College of Science-King, Saud UniversityFaculty of Science, Zarqa UniversityUPMC Hillman Cancer Center, Division of Hematology and Oncology, Department of Medicine, University of PittsburghDepartment of Food Sciences & Nutrition, College of Food and Agricultural Sciences, King Saud UniversityDepartment of Biochemistry and Biotechnology, The Islamia University of BahawalpurAbstract Background Thyroid cancer, characterized by significant genetic and epigenetic alterations, remains a critical focus of molecular oncology. This study investigates eight key genes (BRAF, EIF1 AX, FOXE1, KRAS, PDGFRA, PIK3 CA, PTEN, and TERT) that are deregulated in Hashimoto's Thyroiditis and their roles in thyroid cancer. Methods Cell culture, nucleic acid extraction, RT-qPCR, bisulfite sequencing, and various in silico tools and databases. Results Expression analysis using RT-qPCR revealed significant (p-value < 0.05) down-regulation of BRAF, EIF1 AX, FOXE1, KRAS, PDGFRA, PIK3 CA, PTEN, and TERT genes in thyroid cancer cell lines compared to controls, with ROC curves indicating high diagnostic accuracy (AUC 0.93–0.99). Bisulfite sequencing demonstrated increased promoter methylation across all eight genes in cancerous samples, suggesting epigenetic silencing as a regulatory mechanism. Validation through UALCAN, OncoDB, and HPA confirmed reduced gene and protein expression in additional thyroid cancer cohorts. Genetic alteration analysis via cBioPortal showed prevalent BRAF mutations, whereas other genes exhibited fewer alterations. Kaplan–Meier survival analysis linked lower expression of BRAF and PIK3 CA to poorer overall survival. Correlation studies using TISIDB and TISCH2 databases highlighted associations between gene expression and immune modulation, revealing significant correlations with immune cell infiltration and diverse immune subtypes. Moreover, miRNA-mRNA network analysis identified hsa-mir- 628 - 5p as a critical regulator targeting these genes. The impact of BRAF overexpression on SW579 cells was assessed through various functional assays. Overexpression of BRAF resulted in reduced cell proliferation, colony formation, and wound healing, which may reflect context-dependent effects. While BRAF is typically oncogenic, its overexpression may lead to cellular stress or negative feedback mechanisms that impair these processes. Conclusion This comprehensive analysis elucidates the complex regulatory landscape of these genes in thyroid cancer, emphasizing the significant role of epigenetic modifications and providing insights into potential diagnostic and therapeutic avenues.https://doi.org/10.1186/s41065-025-00429-0Thyroid cancerHashimoto’s ThyroiditisDiagnosisPrognosisTreatment
spellingShingle Mostafa A. Abdel-Maksoud
Taghreed N. Almana
Saeedah Almutair
Abdulaziz Alamri
Ibrahim A. Saleh
Mohamed Y. Zaky
Wahidah H. Al-Qahtani
Yasir Hameed
Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
Hereditas
Thyroid cancer
Hashimoto’s Thyroiditis
Diagnosis
Prognosis
Treatment
title Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
title_full Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
title_fullStr Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
title_full_unstemmed Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
title_short Deciphering the role of Hashimoto's Thyroiditis-related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
title_sort deciphering the role of hashimoto s thyroiditis related key genes in thyroid cancer via detailed in silico analysis followed by the experimental validation
topic Thyroid cancer
Hashimoto’s Thyroiditis
Diagnosis
Prognosis
Treatment
url https://doi.org/10.1186/s41065-025-00429-0
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