The Novel Member of the BCL2 Gene Family, BCL2L12, Is Substantially Elevated in Chronic Lymphocytic Leukemia Patients, Supporting Its Value As a Significant Biomarker

Abstract

BCL2L12 is a recently identified gene belonging to the BCL2 family, members of which are implicated in hematologic malignancies, including chronic lymphocytic leukemia (CLL). The aim of this study was to analyze the mRNA expression of the novel apoptosis-related gene BCL2L12 in patients with CLL and to examine its prognostic and predictive value and potential clinical application as a novel molecular biomarker for CLL. For this purpose, total RNA was isolated from peripheral blood of 65 CLL patients and 23 healthy donors. An ultrasensitive quantitative real-time polymerase chain reaction methodology for BCL2L12 and BCL2 mRNA quantification was developed using SYBR Green chemistry. After preparing cDNA by reverse transcription, relative quantification analysis was performed using the comparative C T (2 ؊⌬⌬CT ) method. Furthermore, analysis of IGHV mutational status, CD38 expression, and detection of early apoptosis by double staining with Annexin V-FITC and propidium iodide were performed. According to our findings, BCL2L12 mRNA expression is significantly higher in CLL patients than in healthy donors. Receiver operating characteristic analysis demonstrated that BCL2L12 expression had significant discriminatory value, distinguishing very efficiently CLL patients from the non-leukemic population. Moreover, BCL2L12 expression predicts the presence of CLL, as demonstrated by both univariate and multivariate logistic regression analyses. Finally, high BCL2L12 mRNA levels are associated with advanced clinical stage and predict shorter overall survival in CLL patients.

Network Biomarkers Constructed from Gene Expression and Protein-Protein Interaction Data for Accurate Prediction of Leukemia

“…To evaluate the performance of network biomarkers in classifying leukemia and normal gene expression profiles, we used three independent gene expression datasets listed in Table 3 as tested datasets to produce the ROC curves. We compared the network biomarker with three reported gene biomarkers: CD38 47 , BCL2 48 and IGFBP7 49 . The reasons we chose these three markers for comparison are as follows, 1) these biomarkers are all well-studied and all of them have been validated by clinical experiments.…”

Molecular cloning of novel alternatively spliced variants of BCL2L12, a new member of the BCL2 gene family, and their expression analysis in cancer cells

“…Recently, we also showed that BCL2L12 mRNA is dramatically elevated in CLL patients, in comparison with healthy controls. Interestingly, BCL2L12 mRNA expression was found to possess significant discriminatory value in CLL, distinguishing very efficiently CLL patients from non-leukemic population, and to constitute an unfavorable prognostic biomarker in CLL, in terms of overall survival (Papageorgiou et al, 2011).…”

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

“…Following the manufacturer's instructions, total RNA was extracted and diluted in a RNA Storage Solution (Ambion Europe Ltd.) and stored at )80°C until use. First-strand cDNA was then synthesized using the M-MuLV Reverse Transcriptase, RNase H ) (Finnzymes Oy, Vantaa, Finland), RNaseOUTÔ RNase inhibitor (Invitrogen, Carlsbad, CA, USA) and oligo(dT) [12][13][14][15][16][17][18] as primer, according to the manufacturer's instructions.…”