Sains Malaysiana 55(8)(2026): 1272-1283

http://doi.org/10.17576/jsm-2026-5508-04

 

Emerging Immunohistochemical Biomarkers Beyond Prostate Specific Antigen in Prostate Cancer

(Kemunculan Penanda Bioimunohistokimia Melangkaui Antigen Khusus Prostat dalam Kanser Prostat)

 

HUMA AFZAL, NOSHEEN ASLAM*, GHULAM MUSTAFA & SHAZIA ANWER BUKHARI

 

Department of Biochemistry, Government College University Faisalabad, Pakistan

 

Received: 13 March 2026/Accepted: 3 August 2026

 

Abstract

Prostate cancer remains a significant health concern, particularly among the older male population. Proper differentiation between malignant adenocarcinoma and benign prostatic diseases is critical for successful diagnosis. This study aimed to determine the dependability of immunohistochemistry (IHC) markers specifically AMACR, p63, and Cytokeratin as a means of diagnosing cancer. The diagnostic value of HOXB13 gene variants and MYC mRNA expression in tissues with benign prostatic hyperplasia (BPH) was also evaluated. The research involved patients with clinical indicators of prostate dysfunction and high PSA values. Tissue specimens were tested using the PIN-4 cocktail (IHC) and Hematoxylin & Eosin (H&E) staining. The ethanol-preserved prostate cancer tissue samples were homogenized to extract total RNA using the Gene JET RNA Purification Kit based on silica membranes and adjusted for high-purity RNA extraction. The concentration and purity of the extracted RNA were evaluated, and only samples exhibiting an A260/A280 ratio within the range of 1.8 to 2.0 were deemed appropriate for downstream analyses. Histopathology confirmed prostatic adenocarcinoma in all cases, mostly low-grade with a minor group of intermediate-grade malignancies. These findings support the notion that IHC markers, specifically AMACR combined with a basal cell marker, are useful modalities for distinguishing between benign and malignant prostate tumors. It also suggests that the rare rs8556 variant is functionally important and potentially deleterious. In prostate cancer treatment, this technique can potentially assist in earlier diagnosis, reduced uncertainty in diagnosis, and personalized treatment.

Keywords: Gene expression; prostate cancer; RNA extraction; Sanger sequencing

Abstrak

Kanser prostat kekal sebagai masalah kesihatan yang utama, terutamanya dalam kalangan populasi lelaki tua. Pembezaan yang betul antara adenokarsinoma malignan dan penyakit prostat benigna adalah penting untuk diagnosis yang berjaya. Kajian ini bertujuan untuk menentukan kebergantungan penanda imunohistokimia (IHC) khususnya AMACR, p63 dan Sitokeratin sebagai cara untuk mendiagnosis kanser. Nilai diagnostik varian gen HOXB13 dan pengekspresan mRNA MYC dalam tisu dengan hiperplasia prostat benigna (BPH) juga dinilai. Penyelidikan ini melibatkan pesakit dengan petunjuk klinikal disfungsi prostat dan nilai PSA yang tinggi. Spesimen tisu diuji menggunakan pewarnaan koktel PIN-4 (IHC) dan Hematoxylin & Eosin (H&E). Sampel tisu kanser prostat yang diawet etanol dihomogenkan untuk mengekstrak RNA total menggunakan Kit Penulenan RNA Gene JET berdasarkan membran silika dan diselaraskan untuk pengekstrakan RNA berketulenan tinggi. Kepekatan dan ketulenan RNA yang diekstrak dinilai dan hanya sampel yang menunjukkan nisbah A260/A280 dalam julat 1.8 hingga 2.0 dianggap sesuai untuk analisis hiliran. Histopatologi mengesahkan adenokarsinoma prostat dalam semua kes, kebanyakannya gred rendah dengan sekumpulan kecil malignan gred pertengahan. Penemuan ini menyokong tanggapan bahawa penanda IHC, khususnya AMACR yang digabungkan dengan penanda sel basal, adalah modaliti yang penting untuk membezakan antara tumor prostat benigna dan malignan. Ia juga menunjukkan bahawa varian rs8556 yang jarang berlaku adalah penting secara fungsi dan berpotensi merosakkan. Dalam rawatan kanser prostat, teknik ini berpotensi membantu dalam diagnosis lebih awal, mengurangkan ketidakpastian dalam diagnosis dan rawatan yang diperibadikan.

Kata kunci: Ekspresi gen; kanser prostat; pengekstrakan RNA; penjujukan Sanger

 

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*Corresponding author; email: nosheenaslam@gcuf.edu.pk