Marcin Słojewski

Smoking related cancers and loci at chromosomes 15q25, 5p15, 6p22.1 and 6p21.33 in the Polish population.

Genetic factors associated with the risk of smoking related cancers have until recently remained elusive. Since the publication of a genome-wide association study (GWAS) on lung cancer new genetic loci have been identified that appear to be associated with disease risk. In this replication study we genotyped 14 single nucleotide polymorphisms (SNPs) located at the 5p12.3-p15.33, 6p21.3-p22.1, 6q23-q27 and 15q25.1 loci in 874 lung, 450 bladder, 418 laryngeal cancer cases and cancer-free controls, matched by year of birth and sex to the cases. Our results revealed that loci in the chromosome region 15q25.1 (rs16969968[A], rs8034191[G]) and 5p15 (rs402710[T]) are associated with lung cancer risk in the Polish population (smoking status adjusted OR = 1.45, 1.35, 0.77; p≤0.0001, 0.0005, 0.002; 95%CI 1.23–1.72, 1.14–1.59, 0.66–0.91 respectively). None of the other regions analyzed herein were implicated in the risk of lung, bladder or laryngeal cancer. This study supports previous findings on lung cancer but fails to show association of SNPs located in 15q25.1 and 5p15 region with other smoking related cancers like bladder and laryngeal cancer.

Microelements in Stones, Urine, and Hair of Stone Formers: A New Key to the Puzzle of Lithogenesis?

The role of trace elements in lithogenesis is still unclear. The aim of this study was to evaluate the levels of elements in urinary stones and in the urine and hair of stone formers to identify these elements that have synergic correlations in studied materials and may contribute to lithogenesis. A total of 219 consecutive patients with idiopathic upper urinary tract stones were prospectively enrolled in the study. Urine and hair samples were collected from all patients. The content of the stone was evaluated using atomic absorption spectrometry, spectrophotometry, and colorimetric methods. The analysis of 29 elements in stones and hair and 21 elements in urine was performed using inductively coupled plasma-atomic emission spectrometry. The strength of correlation was described with the value of Spearman’s rank correlation coefficient. The positive correlation between concentration of sodium, potassium, magnesium, barium, vanadium, zinc, silicon, phosphorus, and iodine in phosphate stones was observed. Only a few incidental correlations between the composition of stones and the distribution of elements in urine and in hair were found. There were 109 positive two-element correlations between two materials. The most common were observed for vanadium, aluminum, lead, cobalt, and molybdenum. Two-element positive correlations for all samples were established only for three elements: vanadium, lead, and aluminum. Results indicate that analysis of particular elements in hair and urine cannot predict the composition of urinary stones. This study showed, for the first time, correlations between the levels of vanadium, lead, and aluminum in the stones, urine, and hair of stone formers.

Germline mutations in the CHEK2 kinase gene are associated with an increased risk of bladder cancer

Germline mutations in CHEK2 have been associated with a range of cancer types but little is known about disease risks conveyed by CHEK2 mutations outside of the context of breast and prostate cancer. To investigate whether CHEK2 mutations confer an increased risk of bladder cancer, we genotyped 416 unselected cases of bladder cancer and 3,313 controls from Poland for 4 founder alleles in the CHEK2 gene, each of which has been associated with an increased risk of cancer at other sites. A CHEK2 mutation (all variants combined) was found in 10.6% of the cancer cases and in 5.9% of the controls (OR = 1.9; 95%CI 1.3–2.7; p = 0.0003). We conclude that CHEK2 mutations increase the risk of bladder cancer in the population.

Does smoking have any effect on urinary stone composition and the distribution of trace elements in urine and stones

The role of particular elements in lithogenesis is still unclear and debated. Probably some of them may promote or conversely inhibit crystal nucleation of organic or mineral species. A few epidemiological data link smoking with the risk of calcium stones. The aim of this hospital-based study was to evaluate the distribution of trace elements in urine and urinary stones, and possible correlation with stone constituents in smoking and non-smoking individuals. 209 stones and urine samples collected from idiopathic stone-formers were analyzed to evaluate the mineral composition and the distribution of elements, 29 in stones and 21 in urine. Values were statistically compared considering smoking, arterial hypertension and coronary heart disease as grouping variables. No differences were noted either for comparison of mineral components or the elements concentrations in stones in both groups. The concentration of mercury in urine was higher in smokers than in non-smokers, but the statistical significance was at the moderate level. Our findings do not support the concept of possible association between smoking and urinary lithogenesis, but we believe that further investigations are needed in this area.

A common nonsense mutation of the BLM gene and prostate cancer risk and survival

BACKGROUND Germline mutations of BRCA2 and NBS1 genes cause inherited recessive chromosomal instability syndromes and predispose to prostate cancer of poor prognosis. Mutations of the BLM gene cause another chromosomal instability clinical syndrome, called Bloom syndrome. Recently, a recurrent truncating mutation of BLM (Q548X) has been associated with a 6-fold increased risk of breast cancer in Russia, Belarus and Ukraine, but its role in prostate cancer etiology and survival has not been investigated yet. METHODS To establish whether the Q548X allele of the BLM gene is present in Poland, and whether this allele predisposes to poor prognosis prostate cancer, we genotyped 3337 men with prostate cancer and 2604 controls. RESULTS Q548X was detected in 13 of 3337 (0.4%) men with prostate cancer compared to 15 of 2604 (0.6%) controls (OR=0.7; 95% CI 0.3-1.4). A positive family history of any cancer in a first- or second-degree relative was seen only in 4 of the 13 (30%) mutation positive families, compared to 49% (1485/3001) of the non-carrier families (p=0.3). The mean follow-up was 49months. Survival was similar among carriers of Q548X and non-carriers (HR=1.1; p=0.9). The 5-year survival for men with a BLM mutation was 83%, compared to 72% for mutation-negative cases. CONCLUSIONS BLM Q548X is a common founder mutation in Poland. We found no evidence that this mutation predisposes one to prostate cancer or affect prostate cancer survival. However, based on the observed 0.6% population frequency of the Q548X allele, we estimate that one in 100,000 children should be affected by Bloom syndrome in Poland.

Nuclear factor E2-related factor-2 (Nrf2) expression and regulation in male reproductive tract

BACKGROUND Nuclear factor E2-related factor-2 (Nrf2, Nfe2l2) plays an important, protective role in many tissues. However, information on molecular mechanisms of detoxification and drug metabolism regulated by Nrf2/NRF2 in testis and epididymis is scarce, but it may help to better characterize the function of blood-testis and epididymis barriers. METHODS Constitutive gene expression was analyzed by real time PCR with TaqMan Assay using ΔCT-method. Additionally, gene expression after treatment with oltipraz- specific Nrf2 inducer was evaluated using ΔΔCT-method. Cellular localization of the Nrf2 was visualized by immunohistochemical reaction. RESULTS The study showed that Nrf2 mRNA level in rat epididymis was higher than in testis. In human tissues, both testis and epididymis demonstrated similar expression levels of NRF2. Immunohistochemical analysis revealed NRF2/Nrf2 protein expression in testis and epididymis, which in the case of testis was dependant on spermatogenesis stage. Both in human and rat tissues constitutive expression of NQO1/Nqo1 was slightly higher in epididymis than in testis. Other Nrf2 regulated genes: GCLC/Gclc and UGT1A6/Ugt1a6 showed different ratios of testis/epididymis/liver expression levels. Treatment with oltipraz (Nrf2 inducer) resulted in significant induction of Nrf2 expression solely in corpus of epididymis. CONCLUSIONS Components of the Nrf2/NRF2 system along with coordinated genes are expressed in testis and epididymis. Moreover, some interspecies differences between rat and human were observed, which may impact extrapolation of experimental data into clinical findings. Studies on animal model showed that corpus of epididymis is the most responsive part of the male reproductive tract to oltipraz exposure at the gene expression level.

Impact of “non-clamping technique” on intra- and postoperative course after laparoscopic partial nephrectomy

Introduction The use of kidney warm ischaemia during laparoscopic partial nephrectomy (LPN) may lead to damage of renal vessels and kidney failure. Laparoscopic partial nephrectomy done without clamping the renal pedicle is feasible and may be beneficial for the postoperative course. Aim To compare intra- and postoperative course in patients undergoing LPN with and without kidney warm ischaemia. Material and methods The material comprises 38 consecutive patients, who underwent LPN in our department during the years 2008-2009. In all cases renal vessels were identified and dissected at first, then resection of the tumour was done. Warm ischaemia was used only in case of difficulties with identification of tumour margin or with the management of bleeding. Out of 38 operations 13 were done without clamping the renal pedicle (group 1) and in the remaining 25 warm ischaemia was applied (group 2). Results Mean dimension of resected tumours in groups 1 and 2 was 31 mm and 33 mm respectively (p > 0.05), while parameters of intra- and postoperative course differed significantly between the groups: mean blood loss – 135 ml vs. 354 ml (p < 0.05), time of surgery – 72.6 min vs. 132.2 min (p < 0.05), postoperative drain leakage – 290 ml vs. 504 ml (p < 0.05), postoperative hospital stay – 3.1 days vs 5.3 days (p < 0.05). In all patients baseline creatinine levels were normal while after surgery creatinine elevation over the upper limit was found in groups 1 and 2 in one and in 6 patients respectively (p < 0.05). Conclusions Laparoscopic resection of kidney tumour without warm ischaemia is feasible and beneficial in pre- and intraoperatively selected cases. Bleeding from renal parenchyma, which requires renal pedicle clamping, may seriously deteriorate intra- and postoperative course in patients undergoing laparoscopic partial nephrectomy.

Evidence-based recommendations on androgen deprivation therapy for localized and advanced prostate cancer

Introduction The management of prostate cancer (PC) is still evolving. Although, androgen deprivation therapy (ADT) is an established treatment option, particularly in patients with disseminated disease, important data regarding hormonal manipulation have recently emerged. The aim of this paper is to review the evidence on ADT, make recommendations and address areas of controversy associated with its use in men with PC. Material and methods An expert panel was convened. Areas related to the hormonal management of patients with PC requiring evidence review were identified and questions to be addressed by the panel were determined. Appropriate literature review was performed and included a search of online databases, bibliographic reviews and consultation with experts. Results The panel was able to provide recommendations on: 1) which patients with localised PC should receive androgen deprivation in conjunction with radiotherapy (RT); 2) what standard initial treatment should be used in metastatic hormone-naïve PC (MHNPC); 3) efficacy of androgen deprivation agents; 4) whether ADT should be continued in patients with castration resistant PC (CRPC). However, no recommendations could be made for combined ADT and very high-dose RT in patients with an intermediate-risk disease. Conclusions ADT remains the cornerstone of treatment for both metastatic hormone-naïve and castration-resistant PC. According to the expert panels opinion, based on the ERG report, luteinizing hormone-releasing hormone agonists might not be equivalent but this needs to be confirmed in long-term data. The combined use of ADT and RT improves outcome and survival in men with high-risk localised disease. The benefits in patients with intermediate-risk disease, particularly those subject to escalated dose RT are controversial.

The presence of prostate cancer at biopsy is predicted by a number of genetic variants

Several single nucleotide polymorphisms (SNPs) have been associated with an elevated risk of prostate cancer risk. It is not established if they are useful in predicting the presence of prostate cancer at biopsy or if they can be used to define a low‐risk group of men. In this study, 4,548 men underwent a prostate biopsy because of an elevated prostate specific antigen (PSA; ≥4 ng/mL) or an abnormal digital rectal examination (DRE). All men were genotyped for 11 selected SNPs. The effect of each SNP, alone and in combination, on prostate cancer prevalence was studied. Of 4,548 men: 1,834 (40.3%) were found to have cancer. A positive association with prostate cancer was seen for 5 of 11 SNPs studied (rs1800629, rs1859962, rs1447295, rs4430796, rs11228565). The cancer detection rate rose with the number of SNP risk alleles from 29% for men with no variant to 63% for men who carried seven or more risk alleles (OR = 4.2; p = 0.002). The SNP data did not improve the predictive power of clinical factors (age, PSA and DRE) for detecting prostate cancer (AUC: 0.726 vs. 0.735; p = 0.4). We were unable to define a group of men with a sufficiently low prevalence of prostate cancer that a biopsy might have been avoided. In conclusion, our data do not support the routine use of SNP polymorphisms as an adjunct test to be used on the context of prostate biopsy for Polish men with an abnormal screening test.

Common variants of xeroderma pigmentosum genes and prostate cancer risk

The genetic basis of prostate cancer (PC) is complex and appears to involve multiple susceptibility genes. A number of studies have evaluated a possible correlation between several NER gene polymorphisms and PC risk, but most of them evaluated only single SNPs among XP genes and the results remain inconsistent. Out of 94 SNPs located in seven XP genes (XPA-XPG) a total of 15 SNPs were assayed in 720 unselected patients with PC and compared to 1121 healthy adults. An increased risk of disease was associated with the XPD SNP, rs1799793 (Asp312Asn) AG genotype (OR=2.60; p<0.001) and with the AA genotype (OR=531; p<0.0001) compared to the control population. Haplotype analysis of XPD revealed one protective haplotype and four associated with an increased disease risk, which showed that the A allele (XPD rs1799793) appeared to drive the main effect on promoting prostate cancer risk. Polymorphism in XPD gene appears to be associated with the risk of prostate cancer.