Determination of expression level of AP1S1, CDK9, FIGF and HDAC11 genes in bladder tumors for aggressive phenotype characterization

AbstractBladder cancer holds a steady 10th place among oncological diseases. Follow-up and timely diagnosis of recurrence and progression of bladder cancer are still based on regular but outdated cystoscopy followed by cytological examination. To reduce the number of cystoscopy procedures, new and reliable biomarkers for predicting tumor behavior must be developed. The aim of this study was to confirm our previous results that demonstrated overexpression of AP1S1, CDK9, FIGF and HDAC11 in muscle-invasive bladder cancer. The project management was performed using iterative flexible project work (Flexible Methodology for Innovative Projects in Scientific Organizations, FMIPSO). Gene expression analysis of the AP1S1, CDK9, FIGF and HDAC11 genes was evaluated in 39 newly collected non-invasive and muscle-invasive bladder tumors. Differential gene expression was calculated using the ΔΔCt method with GPDH as a housekeeping gene. The 4,0-fold change in gene expression was used as a cutoff to determine up- or down-regulation compared to the negative control. Our results demonstrate the involvement of the FIGF, CDK9 and HDAC11 in tumor progression and their potential use as candidate biomarkers to characterize invasive tumor phenotype and muscle progression, as well as potentially reduce the number of cystoscopies. We used FMIPSO to be able to achieve the results at the optimum level of efficiency and control of the project with all possible constraints in scientific organizations

Analysis of the expression level of genes associated with the metabolism of exogenous substances and DNA reparation in patients suffering from bladder cancer

IntroductionBladder cancer is a result of the interaction between urine metabolites that come into contact with the bladder mucosa and a certain hereditary predisposition. AimThe aim of the study is to determine the expression level of genes, involved in the metabolism of xenobiotics that are metabolized to substances with carcinogenic potential, as well as the expression levels of genes involved in DNA reparation.Materials and Methods A total of 16 samples from transitional cell bladder cancer were analyzed - four stage pTa, four pT1, five pT2 and three control samples. Expression analysis was done via PAHS-004 Human Cancer Drug Resistance & Metabolism PCR Array on 29 genes, known to take part in DNA reparation and biotransformation of the xenobiotics: APC, ATM, BRCA1, BRCA2, ERCC3, MSH2, XPA, XPC, ARNT, BLMH, CLPTM1L, CYP1A1, CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A5, DHFR, EPHX1, GSK3A, GSTP1, NAT2, SOD1, SULT1E1 (STE) and TPMT. Increased/decreased expression levels were defined by a fivefold change.Results None of the reparative genes showed increased expression. XPA and APC had a decreased expression, ranging from 6 to 30 times, in the highest number of samples: 11 of 13 for XPA and 10 of 13 for APC.Genes related to metabolism of xenobiotics also showed normal or decreased expression. Only three samples from pTa stage showed up-regulation of the CYP3A5 gene, ranging from 6 to 11 times. The rest of the samples showed normal expressions. Only one sample - low differentiated T2 bladder tumor, grade II, stage G3 showed an increased expression of the genes CLPTM1L, CYP1A1 and GSTP1

Expression analysis of bladder tumors for chemotherapeutic drug sensitivity determination

Introduction: Bladder cancer is a multifactorial disease with increasing frequency in the economically developed countries. Aim: The primary endpoint of this study was the evaluation of the genetic image of bladder tumors in connection to common anticancer drugs.Materials and Methods: A total of 50 samples were analyzed. 41 samples were from transitorial cell bladder cancer (stages pTa, pT1 and pT2), 6 samples were from chronic inflammatory process (precancerous) and three - negative controls. The gene expression analysis of 168 genes were carried out with two panels for Cancer Drug Resistance & Metabolism PCR Array, Qiagene (84 genes) and PAHS-507 Z - Human Cancer Drug Targets PCR Array, Qiagene (84 genes).Results: The results showed significant up-regulation of the genes: CYP1A1, CYP3A5, AR, CLPTM1L, CCNE1, MVP, TOP2B, AHR and PPARG in the bladder cancer samples compared to the negative control. A statistically significant difference (p <0.0001) was found in the expression levels of EGFR, ERBB2, ERBB4, ABCC1, ABCC3, ARNT, CYP1A1, CYP3A5, EPHX1, MVP and PPARG genes in muscle invasive (pT2) versus non-invasive bladder tumors (pTa and pT1). These genes are involved in the formation of multi-drug resistance and in the metabolism of steroid hormones, cyclosporins, polycyclic aromatic hydrocarbons, as well as some anticancer drugs like Vincristine, Taxol and Thiopurine. The obtained data show the significance of the genes as possible targets in clinical trials for the treatment of bladder cancer

Screening of pharmacogenetic variants associated with drug sensitivity in patients with papillary thyroid carcinoma using next generation sequencing

Thyroid cancer is the most common malignant tumour of the endocrine system. One of the most frequent types of thyroid malignancy is papillary carcinoma. In our study, we performed next generation sequencing (NGS) using a cancer panel (Illumina; Illumina, San Diego, USA) to screen for pharmacogenetic susceptibility variants in blood samples of 10 patients with papillary thyroid cancer (PTC). We report variants rs1042522 (TP53), rs2228001 (XPC), rs2227983 (EGFR), rs13181 (ERCC2), rs17655 (ERCC5) and rs1799939 (RET), which were detected in the analyzed patients either in homozygous and/or heterozygous state previously known to be connected with pharmacogenetic sensitivity to certain drugs in oncology. The results showed the TruSight Cancer Panel to be a useful clinical tool for determination of oncotherapy-associated pharmacogenetic variants in the blood of patients

Novel genes and variants associated with longevity in Bulgarian centenarians revealed by whole exome sequencing DNA pools: a pilot study

Aim: To determine specific genetic loci that might be associated with longevity in Bulgarian population by analyzing exome pool-seq data from centenarians and a control group.Methods: We performed whole-exome sequencing of two DNA pools, set up of 32 Bulgarian centenarians and 61 young healthy controls, respectively, and 59935 quality filtered variants were concurrently detected in both pools. Fisher’s exact test was employed to establish the significance of allele frequency difference between the pools.Results: Forty seven variants showed significantly higher allele frequency in the centenarian compared to the control pool, and these can be considered to be positively associated with longevity in Bulgarian populaton. Based on their assigned functional role, three genes containing three of these variants were further investigated. These genes, RNF43, WNK1 and NADSYN1, are involved in evolutionary conserved processes with well ascertained association with longevity, i.e., Wnt signaling pathway, insulin/IGF-1 signal pathway and redox balancing processes, respectively. For the remaining genes exhibiting variants with significantly higher allele frequency in the Bulgarian centenarian pool there is not enough evidence about their functional role in determining longevity and further research is needed.Conclusion: The results confirm the importance of studying centenarians in different populations to discover those combinations of variants that associate with longer health span