Quantitative expression analysis of the apoptotic gene BCL2L12 in breast cancer: association with clinical and molecular prognostic parameters

Apoptosis is a highly orchestrated, genetically regulated form of cell death, the impairment of which is crucial in breast cancer (BC) development and progression. BCL2L12, a member of the BCL2 family of apoptosis-related genes, has been studied in various malignancies, revealing its potential role as a tumor biomarker. It has been recently found that BCL2L12 is subjected to alternative splicing, resulting in the generation of 13 alternatively spliced variants. The aim of this study was the quantification of BCL2L12 splice variants 1 and 2 (v.1 and v.2) expression at the mRNA level and the assessment of their biomarker potential in BC

BCL2L12: A multiply spliced gene with independent prognostic significance in breast cancer

Alternative splicing is a key process in carcinogenesis and, from a clinical aspect, holds great promises, as alternatively spliced variants have emerged as an untapped source of diagnostic and prognostic markers. Our aim was to assess the prognostic value of three recently recognized splice variants of the apoptosis-related gene, BCL2L12, in breast cancer (BC). Total RNA was extracted from breast samples (150 BC and 80 tumor-adjacent normal tissues) and, following cDNA synthesis, a variant-specific qPCR was performed for the expressional quantification of BCL2L12 v.1, v.2 and v.4 transcript variants. Extensive statistical analysis, including bootstrap resampling and internal validation, was conducted in order to evaluate the associations of v.1, v.2 and v.4 expression with patients' clinopathological and survival data. All examined BCL2L12 variants were significantly upregulated in BC specimens compared to their non-cancerous counterpart (v.1, p<0.001; v.2, p=0.009; v.4, p=0.004). Increased BCL2L12 v.4 mRNA expression was associated with markers of unfavorable prognosis namely, advanced tumor grade (p=0.002), ER-(p=0.015)/PR-(p<0.001) negativity, Ki-67-positivity (p=0.007) and high NPI (Nottingham prognostic index) score (p=0.033). Moreover, v.4 was significantly overexpressed in women with triple negative BC (TNBC) and HER2-positive tumors compared to those harboring luminal tumors (p<0.001). Survival analysis disclosed that BCL2L12 v.2 overexpression, as a continuous variable ([HR]=0.45, 95% CI=0.17-0.82, p=0.010), is a strong and independent marker of favorable prognosis for BC patients. Interestingly, v.2 retains its prognostic value in patients with Grade II/III ([HR]=0.21, 95% CI=0.05-0.57, p=0.006) or HER2-positive/TNBC tumors ([HR]=0.25, 95% CI=0.05-0.74, p=0.042). BCL2L12 v.1, v.2, v.4 are aberrantly expressed in BC. Their expressional analysis by cost-effective molecular methods could provide a novel molecular tool for BC management. © 2019 Walter de Gruyter GmbH, Berlin/Boston

Expressional profiling and clinical relevance of RNase κ in prostate cancer: a novel indicator of favorable progression-free survival

Purpose: Considering the unmet need for novel molecular tumor markers capable of improving prostate cancer (CaP) patients’ management along with the fruitful results regarding the future use of ribonucleases (RNases) as molecular diagnostic and prognostic markers in CaP, we aimed to study the expressional profile of RNase κ in CaP and BPH and to investigate its clinical significance in CaP. Methods: Total RNA was extracted from 212 prostatic tissue samples (101 BPH and 111 CaP) and, following cDNA synthesis, quantitative real-time PCR (qPCR) was performed for the expressional quantification of RNase κ. Extensive statistical analysis, including bootstrap resampling, was performed to investigate the differential expression of RNase κ in patients with BPH and CaP and its associations with patients’ clinicopathological and survival data. Results: RNase κ was significantly downregulated (P = 0.002) in CaP patients compared to BPH ones. RNase κ overexpression was associated with decreased risk of CaP development and can discriminate between CaP and BPH independently of serum PSA levels (crude odds ratio = 0.93, P = 0.001). RNase κ upregulation was also associated with less advanced (P = 0.018) and less aggressive (P = 0.001) tumors as well as with longer progression-free survival (PFS) (P = 0.003). Finally univariate bootstrap Cox regression confirmed that RNase κ was associated with favorable prognosis (HR = 0.85, P = 0.002). Conclusions: RNase κ is a biomarker of favorable prognosis in CaP, which is significantly associated with less advanced and aggressive disease, as well as with enhanced PFS. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature

Long noncoding RNAs in digestive system malignancies: A novel class of cancer biomarkers and therapeutic targets?

High throughput methodologies have revealed the existence of an unexpectedly large number of long noncoding RNAs (lncRNAs). The unconventional role of lncRNAs in gene expression regulation and their broad implication in oncogenic and tumor suppressive pathways have introduced lncRNAs as novel biological tumor markers. The most prominent example of lncRNAs application in routine clinical practice is PCA3, a FDA-approved biomarker for prostate cancer. Regarding digestive system malignancies, the oncogenic HOTAIR is one of the most widely studied lncRNAs in the preclinical level and has already been identified as a potent prognostic marker for major malignancies of the gastrointestinal tract. Here, we provide an overview of recent findings regarding the emerging role of lncRNAs not only as key regulators of cancer initiation and progression in colon, stomach, pancreatic, liver, and esophageal cancers, but also as reliable tumor markers and therapeutic tools. lncRNAs can be easily, rapidly, and cost-effectively determined in tissues, serum, and gastric juice, making them highly versatile analytes. Taking also into consideration the largely unmet clinical need for early diagnosis and more accurate prognostic/predictive markers for gastrointestinal cancer patients, we comment upon the perspectives of lncRNAs as efficient molecular tools that could aid in the clinical management. Copyright © 2015 Athina Kladi-Skandali et al

BCL2L12: A promising molecular prognostic biomarker in breast cancer

Objectives: BCL2-like 12 ( BCL2L12) is a new member of the BCL2 gene family that was discovered and cloned by members of our group and found to be expressed in the mammary gland. Many genes of the BCL2 family were found to be implicated in breast carcinogenesis and to serve as possible prognostic markers. The aim of the present study was the quantification of BCL2L12 mRNA expression in order to assess its value as a prognostic tissue biomarker in breast cancer (BC). Design and methods: BCL2L12 mRNA levels were determined in a statistically significant sample size of cancerous (N=108) and adjacent non-cancerous (N=71) breast tissues using a highly sensitive quantitative real-time polymerase chain reaction (qRT-PCR) method. Relative quantification analysis was conducted using the comparative CT (2-δδCT) method, whereas the association between BCL2L12 expression and clinopathological data, disease-free survival (DFS) and overall survival (OS) were estimated by statistical analysis. Results: BCL2L12 mRNA expression was decreased in malignant samples compared to the histologically normal counterparts ( p= 0.012). Significant relationships between BCL2L12 expression and TNM stages ( p= 0.009), metastatic potential ( p= 0.012), tumor size ( p= 0.04) and age ( p= 0.024) were observed. Moreover, Kaplan-Meier and Cox univariate analyses indicated that BCL2L12 expression is associated with longer DFS, whereas multivariate analysis pointed out the independent favorable prognostic value of BCL2L12. Conclusions: According to our results, BCL2L12 mRNA expression is a favorable prognostic marker of DFS for BC patients, suggesting its possible application as a novel prognostic indicator of this malignancy. © 2014

Exercise-induced oxidative stress in G6PD-deficient individuals.

PURPOSE: This study was designed to investigate whether individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency can exercise without greater perturbations in their redox status compared with non-G6PD-deficient individuals. METHODS: Nine males with established G6PD deficiency and nine males with normal G6PD activity performed two exhaustive treadmill exercise protocols of different duration (the shorter one lasting 12 min and the longer one 50 min). Several hematological parameters, reduced glutathione (GSH), oxidized glutathione (GSSG), thiobarbituric acid reactive substances (TBARS), protein carbonyls, catalase, and total antioxidant capacity (TAC) were measured in the blood before and after each exercise bout. RESULTS: Both GSH and GSSG were significantly higher in the control group compared with the G6PD-deficient group at baseline (0.404 +/- 0.101 vs 0.195 +/- 0.049 mmol.L(-1) for GSH and 0.047 +/- 0.012 vs 0.012 +/- 0.006 mmol.L(-1) for GSSG; P 0.05). All other oxidative stress indices were not different between groups at rest (P > 0.05). Exercise of both durations affected significantly (P < 0.05) and similarly the levels of all oxidative stress indices either in the G6PD-deficient group or in the control group. Only the long exercise affected GSH status significantly (P < 0.05), whereas both short and long exercise increased the levels of TBARS, protein carbonyls, catalase activity, and TAC to a similar extent (P < 0.05). CONCLUSION: G6PD-deficient individuals are able to exercise until exhaustion without higher oxidative stress compared with non-G6PD-deficient individuals. Exercise duration is an important determinant of the magnitude of exercise-induced changes for GSH, GSSG, and GSH/GSSG, but not for TBARS, protein carbonyls, catalase activity, or TAC

Prognostic impact of indoleamine 2,3-dioxygenase 1 (IDO1) mRNA expression on circulating tumour cells of patients with head and neck squamous cell carcinoma

Background We sought to determine the prognostic role of indoleamine 2,3-dioxygenase 1 (IDO1) by evaluating IDO1 expression in circulating tumour cells (CTCs) at baseline and after completion of chemoradiotherapy in patients with locally advanced (LA) head and neck squamous cell carcinoma (HNSCC) treated with curative intent. Methods In a prospective cohort of 113 patients with LA HNSCC, we evaluated expression of IDO1 in the EpCAM+ CTC fraction at baseline and after cisplatin chemoradiation. The prognostic value of combined programmed cell death ligand-1 (PDL-1) and IDO1 expression was assessed. Results IDO1 was significantly overexpressed at baseline compared with the post-treatment counterparts (p=0.007). IDO1 messenger RNA (mRNA) expression at baseline was associated with better survival in terms of progression-free survival (PFS) (HR=0.19, p=0.017). Post-treatment IDO1 mRNA levels were correlated with unfavourable prognosis in terms of overall survival (OS) (HR=3.27, p=0.008). Patients with combined decreased expression levels of PDL-1 and IDO1 after treatment exhibited superior PFS (p=0.043) and OS (p=0.021). Conclusions Our results strongly suggest that IDO1 mRNA expression is an independent prognostic factor for clinical outcome. Our study provides useful information for future trials combining chemoradiation with immune checkpoint inhibitors and IDO1 inhibitors. © © Author (s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. Published by BMJ on behalf of the European Society for Medical Oncology