No abstract

IntroductionProstate carcinoma (PRAD) is one of the most frequently diagnosed malignancies amongst men worldwide. It is well-known that androgen receptor (AR) plays a pivotal role in a vast majority of prostate tumors. However, recent evidence emerged stating that estrogen receptors (ERs) may also contribute to prostate tumor development. Moreover, progression and aggressiveness of prostate cancer may be associated with differential expression genes of epithelial-to-mesenchymal transition (EMT). Therefore we aimed to assess the significance of receptors status as well as EMT marker genes expression among PRAD patients in accordance to their age and Gleason score.Materials and methodsWe analyzed TCGA gene expression profiles of 497 prostate tumor samples according to 43 genes involved in EMT and 3 hormone receptor genes (AR, ESR1, ESR2) as well as clinical characteristic of cancer patients. Then patients were divided into four groups according to their age and 5 groups according to Gleason score. Next, we evaluated PRAD samples according to relationship between the set of variables in different combinations and compared differential expression in subsequent groups of patients. The analysis was applied using R packages: FactoMineR, gplots, RColorBrewer and NMF.ResultsMFA analysis resulted in distinct grouping of PRAD patients into four age categories according to expression level of AR, ESR1 and ESR2 with the most distinct group of age less than 50 years old. Further investigations indicated opposite expression profiles of EMT markers between different age groups as well as strong association of EMT gene expression with Gleason score. We found that depending on age of prostate cancer patients and Gleason score EMT genes with distinctly altered expression are: KRT18, KRT19, MUC1 and COL4A1, CTNNB1, SNAI2, ZEB1 and MMP3.ConclusionsOur major observation is that prostate cancer from patients under 50 years old compared to older ones has entirely different EMT gene expression profiles showing potentially more aggressive invasive phenotype, despite Gleason score classification.

IntroductionNotch signalling, an evolutionarily conserved mechanism of cellular differentiation and tissue remodelling, is frequently deregulated in several human malignancies, including renal cell carcinoma (RCC). However, the prognostic value of individual Notch pathway members in RC subtypes remains indefinable. The present study investigates whether the differential expression of Notch members has a contrary effect on disease-free survival (DFS) in clear cell renal cell carcinoma (KIRC), papillary cell renal cell carcinoma (KIRP) and chromophobe renal cell carcinoma (KICH) patients.Material and methodsThe predictive value of 19 Notch members was evaluated in KICH, KIRC and KIRP patient cohorts from The Cancer Genome Atlas (TCGA). Results in the form of Kaplan-Meier survival plots with the p-value calculated (log-rank test, p < 0.05) enabled the patients to be split into favourable/unfavourable prognosis groups regarding expression of Notch members.ResultsMore specifically, lowered expression of ADAM17 correlated with good prognosis in KICH, KIRC and KIRP (HR = 7.79, p = 0.03; HR = 3.98, p = 0.051; HR = 11.24, p < 0.001, respectively). Additionally, HES4 differentiated KICH and KIRC, as its higher expression correlated with good prognosis in KICH and favourable lowered expression in KIRC (HR = 0.11, p = 0.015; HR = 2.42, p < 0.001, respectively).ConclusionsOur analysis could be valuable for better understanding of the molecular mechanism of renal carcinoma. The expression of Notch pathway members could be a useful biomarker for predicting favourable/unfavourable prognosis in patients with RCC.

Breast cancer (BC) is characterized by high heterogeneity regarding its biology and clinical characteristics. The Notch pathway regulates such processes as organ modeling and epithelial-to-mesenchymal transition (EMT). The aim of the study was to determine the effect of differential expression of Notch members on disease-free survival (DFS) in luminal type A (lumA) and triple negative (TN) BC. The differential expression of 19 Notch members was examined in a TCGA BC cohort. DFS analysis was performed using the log-rank test (p<0.05). Biological differences between DFS groups were determined with Gene Set Enrichment Analysis (GSEA) (tTest, FDR<0.25). Common expression profiles according to Notch signaling were examined using ExpressCluster (K-means, mean centered, Euclidean distance metric). The overexpression of HES1, LFNG and PSEN1 was found to be favorable for DFS in lumA, and lowered expression favorable for DFS in TN. GSEA analysis showed that differential Notch signaling is associated with cell cycle, tissue architecture and remodeling. Particularly, targets of E2F, early stage S phase transcription factor, were upregulated in the lumA unfavorable group and the TN favorable group differentiated on a basis of HES1 and PSEN1 expression. Summarizing, our analysis show significance of Notch signaling in BRCA progression through triggering EMT. Moreover, identification of numerous genes which overexpression is associated with disease recurrence may serve as a source of potential targets for a new anticancer therapy.

Uterine corpus endometrial carcinomas (UCEC) are clinically divided into two subgroups—endometrioid endometrial carcinoma (EEC) or non-endometrioid endometrial carcinoma (NEEC). The first group shows relatively better prognosis. However, the discrimination seems to be insufficient due to the fact that in the mildest EEC are patients with poor treatment response and bad prognosis. Our aim was to examine the molecular background of such phenomenon and whether gene expression patterns might be of importance for the clinic. We focused our analysis on WNT pathway target genes since it is one of the main regulators of endometrial proliferation and differentiation. In silico analysis of TCGA data, including Weighted Co-expression Network Analysis, Principle Component Analysis, and Multiple Factor Analysis, allows to select 28 genes that serve as a predictive markers for UCEC patients. Our study revealed that there is a subgroup of the endometrioid cases that molecularly resembles mixed/serous groups. This may explain the reason for existence of subgroup of patients, that although clinically diagnosed with the mildest endometrioid UCEC type, yet present failure in treatment and aggressive course of the disease. Our study suggests that worse outcome in these patients may be based on a disruption of proper WNT signalling pathway resulting in deregulation of its effector genes. Moreover, we showed that mixed group consisting of tumours containing both endometrioid and serous types of cells, has serous expression profile of WNT targets. The proposed gene set allows to predict progression of the disease trough dividing patients into groups of low or high grade with 70.8% sensitivity and 88.6% specificity (AUC = 0.837) as well as could predict patient prognosis associated with UCEC subtype with 70.1% sensitivity and 86.2% specificity (AUC = 0.855). Relatively small number of implicated genes makes it highly applicable and possibly clinically simple and useful tool.

Introduction Prostate cancer (PC) is the first cause of fatality related to the reproductive system in men. One out of seven men encounter this pathology once in their lifetime. Prostate Specific Antigen along with digital rectal exam are the first line of screening for prostate pathologies but numerous studies showed that these techniques lack specificity and in sometimes validity for screening. From this notion comes the need to find new biomarkers and molecular players that help get an early detection and effective treatment of PC. Casein Kinase 1 Alpha (CSNK1A1) is an enzyme involved in multiple cellular processes such as the regulation of the oncogenic Wnt/beta-catenin signalling pathway. Its implication in carcinogenesis and cancer progression is still unclear. In this study, we aim to establish a CSNK1A1 expression profile in BPH and in differently advanced PC grades and to investigate the localization of the enzyme compared to beta catenin in the different samples. Material and methods Formalin-fixed paraffin-embedded human prostatic tissues were collected as follows: 85 BPH, 64 PC and 3 controls. Gene expression was assessed by quantitative RT-PCR of cellular transcripts. Protein localization was monitored by tissue immunostaining for CSNK1A1 and Beta-Catenin. Further in-vitro validation of the results is to be made on normal prostate epithelial cell line and PC cell lines using RT-PCR and western blotting. Results and discussions We were able to identify a continuous increase in the CSNK1A1 expression between controls, BPH and PC. These results are being validated in vitro. Preliminary immunostaining results showed membranous beta catenin in BPH and that seemed to be more heterogeneous in PC sections. A correlation with CSNK1A1 localization and amount is still to be achieved. Our results suggest a role of CSNK1A1 in the pathogenesis of BPH and PC by working on the proliferation induction axis, and malignant behaviour of cells. Our staining results suggest a minor role of beta catenin and Wnt pathway in these axes from which emerges the need to deeply investigate the pathways in which CSNK1A1 is implicated in BPH and PC initiation and progression. Conclusion Our results suggest an increase in CSNK1A1 in BPH and PC, which flags this protein as a potential marker in the progression of PC. More investigation is needed to verify whether this protein is involved in cancer initiation or in the direct inhibition of the oncogenic Wnt/beta-catenin signalling pathway and other pathways.

IntroductionLung carcinomas are very aggressive malignancies, which remain leading cancer-related cause of death among men and women worldwide. Regarding histological classification LUSC and LUAD account for the majority of lung tumours among non-small cell carcinomas (NSCLC). Notch signalling is evolutionarily-conserved pathway that regulates many cellular processes such as proliferation, differentiation and epithelial-to-mesenchymal transition (EMT). To date, aberrant Notch signalling has been linked with wide variety of malignancies, including lung tumours. However the association between co-expression profile of Notch downstream effector and lung cancer subtypes remains unclear. Therefore the aim of our study was to investigate functional gene co-expression networks to search for candidate biomarkers or therapeutic targets.Material and methodsIn our analysis we used RNASeq expression and clinical data downloaded from The Cancer Genome Atlas (TGCA). Gene Set Enrichment Analysis (GSEA) performed for canonical pathways pointed to Notch pathway as one of the most significant. Subsequently we analysed expression of downstream Notch effectors, 2949 HES/HEY transcription factors targets. To this extent used Weighted Gene Co-expression Network Analysis (WGCNA) to find differences in gene expression profile between LUSC and LUAD.Results and discussionsThe analysis of Notch pathway downstream targets which expression is regulated by HES/HEY transcription factors, identified 9 gene modules highly correlated with cancer type, with two of them as the most promising. Functional analysis revealed that among the differentially expressed genes were those involved in proliferation, cell cycle regulation, DNA repair, EMT, adhesion and metabolic processes, for example: TP63, PIK3CA, ADAM23, DLG1, FXR1, SENP5, TTK, BIRC5, KIF18A, KIF14, KIF4A, MCM4, MCM10. For one module the highly connected hub gene is TP63, acts as oncogene in many tumour types including squamous cell carcinomas, and for the second module is KIF4A. Interestingly, all listed genes were found to be overexpressed in LUSC and downexpressed in LUAD.ConclusionOur analysis could be valuable for better understanding of the molecular mechanism of lung carcinoma as well as Notch signalling in lung cancer with emphasis of pathway gene expression as useful biomarker for differentiating cancer progression in lung cancer subtypes.AcknowledgmentThis study was founded by the National Science Centre, Poland nr 2016/23/N/NZ2/02575 and Medical University of Lodz 503/0-078-02/503-01-004

Introduction WWOX gene is a tumour suppressor which loss of expression is demonstrated in many cancers. WWOX is involved in the regulation of glucose metabolism by interaction with HIF1a. WWOX knockdown regulates the metabolic switch from oxidative phosphorylation to aerobic glycolysis in breast cancer MCF7 cell line. This metabolic reprograming called the Warburg effect enhances glycolysis to generate energy over oxidative phosphorylation in aerobic conditions. We considered that aerobic glycolysis can be upregulated via changes in WWOX expression in case of women with gestational diabetes mellitus (GDM). We hypothesised that WWOX has a function in regulating glucose metabolism in GDM. Literature indicates that GDM may be associated with increased risk of breast cancer. Material and methods We analysed expression level of WWOX, HIF1A, glucose transporters SLC2A1, SLC2A4 and glycolytic genes HK2, PFK1, PKM2, LDHA by RT-qPCR in GDM and NGT blood samples. The expression of same genes, we analysed in normal/tumour pair of breast tissue from GEO database (GSE109169). The results, we referred to expression level of these genes in MCF7 and MCF7 sh WWOX cell line obtained by Abu-Remaileh et. al. Results and discussions Relative expression level of WWOX was lower in case of GDM than NGT. Opposite result we observed for HIF1A. Moreover, WWOX/HIF1A ratio is significantly lower in GDM than NGT. The increased level of HIF1A in GDM group is further associated with higher expression of glucose transporters and glycolytic genes. In GDM we observed negative correlation between WWOX/ HIF1A ratio and HIF1A, SLC2A1 as well as HK2, PFK and PKM2. WWOX knockout in MCF7 leads to increasing of HIF1a target glycolytic genes. Similar to GDM, WWOX expression level is lower and HIF1A is higher in breast cancer in comparison to normal breast tissue, which gives statistically lower WWOX/HIF1A ratio. Expression level of SLC2A1, HK2, PKM2, LDHA in breast cancer are higher than in normal. Conclusion We found elevated WWOX and HIF1A expression level, enhanced glycolysis, reduced WWOX/HIF1A ratio in GDM. Our results suggest that WWOX is responsible for Warburg effect occurrence in GDM via regulation of HIF1A, as like in breast cancer. Therefore, a hypothesis can be formed that elevated risk of breast cancer in diabetes mellitus patients could be associated with systemic WWOX function insufficiency.