hitit medical journal Hitit Medical Journal 2687-4717 Hitit Üniversitesi 10.52827/hititmedj.1351295 Pathology Gene and Molecular Therapy Patoloji Gen ve Moleküler Tedavi The Comparison of Real-Time PCR and Mutation-Specific Immunohistochemistry in EGFR Mutation Analysis of Non-Small Cell Lung Carcinomas Küçük Hücreli Dışı Akciğer Karsinomlarında EGFR Mutasyon Analizinde Real-Time PCR ve Mutasyon Özgü İmmünohistokimya Karşılaştırması https://orcid.org/0000-0002-4541-793X Aydın Meriçöz Çisel KOÇ ÜNİVERSİTESİ, TIP FAKÜLTESİ https://orcid.org/0000-0001-6112-8719 Güler Gaye HACETTEPE ÜNİVERSİTESİ, TIP FAKÜLTESİ https://orcid.org/0000-0002-5523-0669 Önder Sevgen HACETTEPE ÜNİVERSİTESİ, TIP FAKÜLTESİ 06 30 2024 6 2 117 125 09 08 2023 04 29 2024 Copyright © 2019, Hitit Medical Journal 2019 Hitit Medical Journal

Objective: This study aims to identify activating mutations in the epidermal growth factor receptor (EGFR) gene in patients with non-small cell lung cancer (NSCLC) and to evaluate their correlation with responses to EGFR-tyrosine kinase inhibitors (TKI) treatment. This study aims to identify activating mutations in the epidermal growth factor receptor (EGFR) gene in patients with non-small cell lung cancer (NSCLC) and to evaluate their correlation with responses to EGFR-tyrosine kinase inhibitors (TKI) treatment. We conducted a comparative analysis of Real-Time PCR and immunohistochemistry to detect EGFR mutation status in non-small cell lung cancer patients, focusing on the sensitivity, specificity, and predictive values of immunohistochemistry.Material and Method: We evaluated 788 non-small cell lung cancer samples which were analyzed for EGFR mutation status by RT-PCR. We detected 126 EGFR mutated cases among these patients. We evaluated mutation-specific EGFR immunohistochemistry directed towards the exon 19 deletions (15 bp E746-A750) and exon 21 point mutation (L858R) to the 47 EGFR mutated patients histologic material and cell blocks of cytologic specimens.Results: 32 of the 47 cases (68%) had exon 19 deletion, 14 of them (30%) had point mutation in exon 21, and one of them (2%) showed exon 18 mutation. EGFR exon 19 (15 bp E746-A750 deletion) antibody showed a sensitivity of 100%, specificity of 40%, negative predictive value of 100%, and positive predictive value of 78%. The sensitivity of the exon 21 (L858R point mutation) antibody was 93%, specificity was 91%, negative predictive value was 97% and positive predictive value was 82%.Conclusion: Our investigation indicates that mutation-specific EGFR immunohistochemistry has demonstrated a notable sensitivity and specificity for exon 21. However, while sensitive, the exon 19 (15 bp E746-A750 deletion) antibody lacked specificity. While positive immunohistochemical staining may suggest the presence of an EGFR mutation, making the patient potentially eligible for TKI treatment, it should not be the sole determinant. If immunohistochemistry results are negative, it is essential to resort to molecular tests to ensure accurate diagnosis and appropriate therapeutic guidance. With evolving diagnostic landscapes, it is crucial to harness both IHC and molecular techniques judiciously for optimal patient care.

Amaç: Bu çalışmanın amacı, Küçük Hücreli Dışı Akciğer Kanseri (KHDAK) hastalarında EGFR genindeki aktivasyon mutasyonlarını tespit etmek ve EGFR-tirozin kinaz inhibitörlerinin (TKI) tedavi yanıtlarıyla olan ilişkisini değerlendirmektir. Küçük hücreli dışı akciğer kanseri (KHDAK) hastalarında epidermal büyüme faktörü reseptörü (EGFR) genindeki aktive edici mutasyonların belirlenmesi, EGFR-tirozin kinaz inhibitörleri (TKI) tedavi yanıtlarıyla ilişkilidir. Mutasyon spesifik antikorlar kullanılarak yapılan immünohistokimya, belirli mutant EGFR proteinlerini tespit edebilmektedir. KHDAK hastalarında EGFR mutasyon durumunu tespit etmek için Real-Time PCR ve immünohistokimyayı karşılaştırdık ve immünohistokimyanın duyarlılık, özgüllük, pozitif ve negatif öngörü değerlerini analiz ettik.Gereç ve Yöntem: RT- PCR ile EGFR mutasyon durumu için analiz edilen 788 küçük hücreli dışı akciğer kanseri örneğini değerlendirildi. Bu hastalar arasında 126 EGFR mutasyonlu vakayı tespit edildi. 47 EGFR mutasyonlu hastanın histolojik materyali ve sitolojik örneklerin hücre bloklarına yönelik ekzon 19 delesyonları (15 bp E746-A750) ve ekzon 21 nokta mutasyonu (L858R) için mutasyon spesifik EGFR immünohistokimyası (IHK) çalışıldı ve sonuçlar boyama kuvveti ve yaygınlığına göre değerlendirildi.Bulgular: 47 vakadan 32’si (%68) ekzon 19 delesyonuna sahipti, bunların 14’ünde (%30) ekzon 21’de noktamutasyonu vardı ve birinde (%2) ekzon 18 mutasyonu gözlendi. EGFR ekzon 19 (15 bp E746-A750 delesyon) antikoru %100 duyarlılık, %40 özgüllük, %100 negatif öngörü değeri ve %78 pozitif öngörü değeri gösterdi. Ekzon 21 (L858R nokta mutasyonu) antikorunun duyarlılığı %93, özgüllüğü %91, negatif öngörü değeri %97 ve pozitif öngörü değeri %82 idi.Sonuç: Araştırmamız, EGFR immünohistokimyasının, ekzon 21 mutasyonu için belirgin bir duyarlılık veözgüllük sergilediğini göstermektedir. Ancak, duyarlı olan ekzon 19 (15 bp E746-A750 delesyon) antikoruözgüllükten yoksundur. Pozitif immünohistokimya sonucu, hastada EGFR mutasyonu olabileceğini ön görebilir ve hasta potansiyel olarak TKI tedavisine uygun olabilir; ancak bu durum tek başına belirleyici olmamalıdır. İmmünohistokimya sonuçları negatifse, doğru tanı ve uygun tedavi rehberliği için moleküler testlere başvurulması esastır.

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