Article

Quantitative expression analysis of the apoptosis-related gene, BCL2L12, in head and neck squamous cell carcinoma

Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Athens, Greece First Ear, Nose and Throat Clinics, Faculty of Medicine, University of Athens, Athens General Hospital 'Hippokration', Athens, Greece.
Journal of Oral Pathology and Medicine (Impact Factor: 1.93). 07/2012; 42(2). DOI: 10.1111/j.1600-0714.2012.01190.x
Source: PubMed

ABSTRACT

J Oral Pathol Med (2013)42: 154–161
Background: BCL2L12 is a recently identified gene belonging to the BCL2 family, members of which are implicated in head and neck squamous cell carcinoma (HNSCC). We have recently shown that BCL2L12 mRNA expression is an unfavorable prognostic indicator in nasopharyngeal carcinoma (NPC) and that BCL2L12 can be regarded as a novel, useful tissue biomarker for the prediction of NPC patients’ short-term relapse. The aim of this study was to analyze the mRNA expression of the novel apoptosis-related gene BCL2L12 in patients with HNSCC.
Methods: Total RNA was isolated from 53 malignant tumors originating in larynx, pharynx, tongue, buccal mucosa, parotid glands, and nasal cavity, as well as from 34 adjacent non-cancerous tissue specimens, resected from patients with HNSCC. A highly sensitive real-time PCR method for BCL2L12 mRNA quantification in head and neck tissues was developed using the SYBR® Green chemistry. After preparing cDNA by reverse transcription, relative quantification was performed using the comparative CT () method.
Results: BCL2L12 mRNA levels were lower in laryngeal tumors of advanced tumor, node, metastasis (TNM) stage or bigger size and in well-differentiated malignant tongue neoplasms, compared with early-stage laryngeal tumors or poorly differentiated tongue tumors. Interestingly, the BCL2L12 expression showed significant discriminatory value, distinguishing efficiently patients with tongue squamous cell carcinoma (SCC) from non-cancerous population.
Conclusions: This is the first study examining the BCL2L12 mRNA expression in HNSCC. Our results suggest that BCL2L12 mRNA expression may serve as a potential prognostic biomarker in tongue and/or larynx SCC, which principally constitute the great majority of HNSCC cases worldwide.

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    • "In addition, cyclin D3 (CCND3) mRNA overexpression and CCND1 gene amplification were shown to independently predict poor OS in this malignancy (Bellacosa et al., 1996; Pruneri et al., 2005). Many other molecules have also been studied in LSCC, including markers of proliferation Ki-67 (MKI67) and proliferating cell nuclear antigen (PCNA) (Franchi et al., 1996; Liu et al., 2003), proapoptotic as well as antiapoptotic BCL2 family proteins (Condon et al., 2002; Geomela et al., 2013), members of the epidermal growth factor receptor (EGFR) family (Ganly et al., 2007; Almadori et al., 2010) and the vascular endothelial growth factor (VEGF) subfamily (Teknos et al., 2002; Sullu et al., 2010), E-cadherin (Franchi et al., 1996), and cathepsins B and D (Maurizi et al., 1996; Li et al., 2011). Furthermore, some microRNAs (miRNAs) appear as emerging tumor biomarkers in LSCC (Saito et al., 2013; Zhao et al., 2013). "
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    ABSTRACT: Abstract Several members of the family of tissue kallikrein and kallikrein-related peptidases have been suggested as promising tumor biomarkers with important prognostic significance. However, only one (KLK11) has already been studied in laryngeal squamous cell carcinoma (LSCC) as potential biomarker for LSCC diagnosis and/or prognosis. Our study investigated the prognostic value of kallikrein-related peptidase-4 (KLK4) mRNA expression as a molecular tissue biomarker in LSCC. For this purpose, KLK4 mRNA expression analysis was performed in 116 cancerous and 74 paired non-cancerous laryngeal tissue specimens obtained from patients having undergone surgical treatment for primary LSCC. A remarkable downregulation of KLK4 mRNA expression was noticed in laryngeal tumors, compared to non-cancerous laryngeal tissue specimens. KLK4 mRNA expression was also shown to distinguish well LSCC from non-cancerous laryngeal tissues. Furthermore, low KLK4 mRNA expression was shown to predict poor disease-free survival, independently of the histological grade and size of the malignant tumor as well as patient TNM stage. According to Kaplan- Meier survival analysis, low KLK4 mRNA expression predicts short-term relapse even among patients with well-differentiated tumors or those at an early TNM stage. Thus, KLK4 mRNA positivity could be regarded as a novel independent indicator of favorable prognosis for the disease-free survival of LSCC patients.
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    • "As illustrated in Fig. 1, the slopes of KLK10 and B2M amplification plots are very similar (−3.350 and −3.403, respectively), which clearly indicates similar efficiencies for the corresponding amplicons (98.8% and 96.7%, respectively). Normalised results were expressed as relative quantification (RQ) units, which stand for the ratio of KLK10 mRNA copies to B2M mRNA copies, calculated for each colorectal tissue specimen, in relation to the same ratio, calculated for the calibrator[51]. The normalised (2 −ΔΔCT ) amounts of each sample KLK10 mRNA levels were then multiplied with the average ratio of KLK10 mRNA copies to B2M mRNA copies of HT-29 cells (2 −6.502 ), which was calculated from the intercept of the regression line. These calculations made our results from different runs comparable and independent of the KLK10 mRNA expression levels of HT-29 cells. "
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