RESEARCH ARTICLE Quantitative analysis of BCL2 mRNA expression in nasopharyngeal carcinoma: an unfavorable and independent prognostic factor Ali Fendri & Christos K. Kontos & Abdelmajid Khabir & Raja Mokdad-Gargouri & Alexandros Ardavanis & Andreas Scorilas Received: 8 March 2010 / Accepted: 26 April 2010 / Published online: 1 June 2010 # International Society of Oncology and BioMarkers (ISOBM) 2010 Abstract Programmed cell death plays a vital role in a wide variety of physiological processes. Defects in apopto- tic cell death contribute to neoplastic diseases by preventing or delaying normal cell death. BCL2 (Bcl-2) is an anti- apoptotic gene with marked up-regulation in various malignancies, such as breast cancer, in which expression of the BCL2 protein has been proposed as a prognostic tumor biomarker. The purpose of the current study was to investigate mRNA expression of the BCL2 gene in nasopharyngeal carcinoma (NPC) biopsies and assess its prognostic value. For this purpose, total RNA was isolated from 89 malignant and hyperplastic nasopharyngeal biop- sies from Tunisian patients. After testing the quality of the extracted RNA, cDNA was prepared by reverse transcrip- tion. A highly sensitive real-time PCR methodology for BCL2 mRNA quantification was developed using SYBR® Green chemistry. GAPDH served as an endogenous control gene. Relative quantification analysis was performed using the comparative C T (2 -ΔΔCT ) method. High BCL2 mRNA levels were detected in advanced-stage nasopharyngeal tumors (p =0.030). Furthermore, BCL2 mRNA expression was strongly associated with lymph node involvement (p = 0.009) and presence of distal metastases (p =0.013). Survival analysis demonstrated that patients with BCL2- positive nasopharyngeal tumors have significantly shorter disease-free and overall survival (p =0.011 and p =0.028, respectively). The major contribution of the current study is the quantification and evaluation, for the first time, of the prognostic significance of the BCL2 mRNA expression in nasopharyngeal carcinoma (NPC) patients. Our results suggest that mRNA expression levels of BCL2 may represent a novel unfavorable and independent tumor biomarker for nasopharyngeal carcinoma. Keywords Apoptosis . BCL2 (bcl-2) . Real-time PCR . Tumor biomarkers . Nasopharyngeal carcinoma (NPC) Introduction Apoptosis is a tightly regulated and finely tuned form of programmed cell death, playing an integral role in normal development, tissue homeostasis, and elimination of dam- aged and infected cells [1]. Disruption of the normal apoptotic process is implicated in a variety of human diseases, such as acquired immunodeficiency syndrome (AIDS), neurodegenerative disorders, autoimmunity, and cancer [2, 3]. Defects in apoptotic mechanisms can prolong the duration of cell life, thus resulting in neoplastic cell expansion independent of mitosis rate [4]. Moreover, A. Fendri : R. Mokdad-Gargouri Unité de Recherche Génétique du Cancer et Production de protéines thérapeutiques, Centre de Biotechnologie de Sfax, Sfax, Tunisia C. K. Kontos : A. Scorilas (*) Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimiopolis, 15701 Athens, Greece e-mail: ascorilas@biol.uoa.gr A. Khabir Centre Hospitalo-Universitaire Habib Bourguiba, Sfax, Tunisia A. Ardavanis First Department of Medical Oncology, “Saint Savvas” Anticancer Hospital, Athens, Greece Tumor Biol. (2010) 31:391–399 DOI 10.1007/s13277-010-0047-3