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


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.
    Full-text · Article · Apr 2014 · Biological Chemistry
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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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    ABSTRACT: Objectives: Colorectal cancer (CRC) is one of the three most common cancers in both genders. Even though several biomarkers are in use in diagnosis and prognosis of the disease, they are marred by limited specificity and sensitivity. The human kallikrein-related peptidase 10 (KLK10) gene is a member of the human tissue kallikrein family. Because prostate specific antigen (PSA), the best biomarker for detecting and monitoring prostate cancer, is a member of this family, many other members, including KLK10, have been widely examined as novel biomarkers for different cancer types. In previous studies, KLK10 has been proposed as a diagnostic biomarker for ovarian carcinoma, while its methylation on exon 3 has been proposed as a prognostic marker for early-stage breast cancer patients. The purpose of this study was to analyse KLK10 mRNA expression and examine its prognostic value and potential clinical application as a novel molecular tissue biomarker in CRC. Design and methods: The study group consisted of 190 colorectal samples. Total RNA was extracted from pulverised tissues and cDNA was prepared by reverse transcription. KLK10 was amplified by real-time PCR. B2M was used as a reference gene and HT-29 cells as positive control. Results: KLK10 expression was significantly higher in cancer tissues (P<0.001). Tumours of advanced TNM and Dukes' stage showed high KLK10 expression status (P=0.036; P=0.025). Patients with high KLK10 expression had a shorter disease-free and overall survival rates (P=0.014; P=0.020). Conclusion: Our results suggest that KLK10 may serve as a new marker of unfavourable prognosis of colorectal cancer.
    Full-text · Article · Mar 2013 · Clinical biochemistry
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    • "In fact, several molecules have been shown to be overexpressed in various malignancies, while being downregulated in other types of cancer. For instance, BCL2L12, a member of the BCL2 family of apoptosis-related genes, is overexpressed in undifferentiated nasopharyngeal carcinoma [43] and in poorly differentiated TSCC, whereas its mRNA levels are lower in LSCC of advanced TNM stage, compared to early-stage laryngeal tumors [45]. "
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    ABSTRACT: Background Head and neck squamous cell carcinoma (HNSCC) represents one of the most commonly diagnosed malignancies worldwide. The DDC gene encodes L-DOPA decarboxylase, an enzyme catalyzing the decarboxylation of L-DOPA to dopamine. We have recently shown that DDC mRNA is a significant predictor of patients’ prognosis in colorectal adenocarcinoma and prostate cancer. The aim of the current study was to analyze the DDC mRNA expression in HNSCC patients. Methods 53 malignant tumors were resected from the larynx, pharynx, tongue, buccal mucosa, parotid glands, and nasal cavity, as well as from 34 adjacent non-cancerous tissues of HNSCC patients, and were homogenized. Total RNA was isolated and converted into first-strand cDNA. An ultrasensitive real-time PCR method based on the SYBR Green chemistry was used for DDC mRNA quantification in head and neck tissue specimens. Relative quantification was performed using the comparative Ct (2-ddCt) method. Results DDC mRNA levels were lower in squamous cell carcinomas (SCCs) of the larynx and tongue than in adjacent non-cancerous tissue specimens. Furthermore, low DDC mRNA expression was noticed in laryngeal and tongue tumors of advanced TNM stage or bigger size, compared to early-stage or smaller tumors, respectively. No statistically significant differences were observed between SCCs resected from pharynx, buccal mucosa, or nasal cavity, and their normal counterparts. Conclusion This is the first study examining the DDC mRNA expression in HNSCC. According to our results, DDC mRNA expression may constitute a potential prognostic biomarker in tongue and/or larynx SCCs, which principally represent the overwhelming majority of HNSCC cases.
    Full-text · Article · Oct 2012 · BMC Cancer
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