Major aim of this project is search for markers of prediction of breast cancer chemotherapy result. Differences in expression of groups of functionally relevant genes will be followed in tumors of patients and compared to chemotherapy outcomes. Revealedpredictive markers will be validated by confirmation of mechanism of action using analysis of genetic polymorphism and phenotype of corresponding proteins. The project will continue in previous studies of principal investigator in the same area.

Cílem projektu je nalezení ukazatelů predikce výsledku chemoterapie u pacientek s karcinomem prsu. V nádorech pacientek budou hledány rozdíly v expresi funkčně důležitých skupin genů a současně bude hodnocena léčebná odpověď na standartní chemoterapii. Nalezené ukazaltele budou validovány pomocí studia mechanismu jejich působení analýzou genetického polymorfismu a fenotypu odpovídajících proteinů. Projekt naváže na řadu studií již rozpracovaných řešitelským pracovištěm.

• MeSH
• cytostatické látky farmakokinetika   MeSH
• exprese genu MeSH
• fenotyp MeSH
• genomika MeSH
• genotyp MeSH
• nádorové biomarkery MeSH
• nádory prsu farmakoterapie   MeSH

• Konspekt
• Farmacie. Farmakologie
• NLK Obory
• farmacie a farmakologie
• onkologie
• gynekologie a porodnictví
• biologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Histologically verified pairs (n=10) of pancreatic tumors and non-neoplastic tissues were used for quantitative real-time PCR and the stability of 24 reference genes was analyzed with geNorm and NormFinder software. Raw C{q} values correlated with the degree of RNA degradation. This correlation was abolished by normalization to C{q} of 18S endogenous control gene. Both geNorm and NormFinder programs suggested EIF2B1, ELF1, MRPL19, and POP4 as the same most stable genes. We have thus identified suitable reference genes for future expression studies in pancreatic carcinoma. Normalization method reducing the effects of RNA degradation on the quality of results was also developed.

• MeSH
• eukaryotický iniciační faktor 2B genetika   MeSH
• exprese genu MeSH
• jaderné proteiny genetika   MeSH
• karcinom diagnóza   genetika   patologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• mitochondriální proteiny genetika   MeSH
• nádorové biomarkery genetika   MeSH
• nádory slinivky břišní diagnóza   genetika   patologie   MeSH
• pankreas metabolismus   patologie   MeSH
• ribonukleasy genetika   MeSH
• ribonukleoproteiny genetika   MeSH
• ribozomální proteiny genetika   MeSH
• senioři MeSH
• software MeSH
• stabilita RNA MeSH
• stanovení celkové genové exprese MeSH
• transkripční faktory genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Associations of transcript levels of oxidative stress-modifying genes SOD2, SOD3, NQO1 and NQO2 and their functional single nucleotide polymorphisms (SNPs) rs4880, rs1799895, rs2536512, rs699473, rs1800566 and rs1143684 with prognosis of breast cancer patients were studied. SNPs were assessed by allelic discrimination in a cohort of 321 breast cancer patients from the Czech Republic. Transcript levels were determined by real-time polymerase chain reaction (PCR) with absolute quantification in tumor and adjacent non-neoplastic control tissues. Both genotypes and transcript levels were then compared with available clinical data on patients. Patients carrying low activity allele Leu in NQO2 rs1143684 had a greater incidence of stage 0 or I disease (i.e., better prognosis) than patients with the Phe/Phe genotype. This association was more evident in patients without expression of progesterone receptors (p = 0.031). Patients carrying the Thr allele in SOD3 rs2536512 SNP had a significantly greater incidence of tumors expressing estrogen receptors than patients carrying the Ala/Ala genotype (p = 0.007). SOD3 transcript level was significantly higher in grade 1 or 2 tumors than in grade 3 tumors (p = 0.006). Patients carrying T allele in SOD3 rs699473 SNP had significantly poorer progression-free survival (PFS) than patients carrying the CC genotype (p = 0.038). The same applied to the subgroup of patients treated by hormonal regimens (p = 0.021). Patients carrying the high activity Ala/Ala genotype in SOD2 (rs4880) had significantly poorer PFS than Val allele carriers in the group treated by cyclophosphamide but not hormonal regimens (p = 0.004). Our results suggest that NQO2, SOD2 and SOD3 may significantly modify prognosis of breast cancer patients and that their significance should be further characterized.

• MeSH
• chinonreduktasy biosyntéza   genetika   MeSH
• DNA nádorová krev   genetika   MeSH
• genetická transkripce MeSH
• jednonukleotidový polymorfismus MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• NAD(P)H dehydrogenasa (chinon) biosyntéza   genetika   MeSH
• nádorové biomarkery biosyntéza   genetika   MeSH
• nádory prsu krev   enzymologie   genetika   patologie   MeSH
• přežití po terapii bez příznaků nemoci MeSH
• superoxiddismutasa biosyntéza   genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: The ABCB1 gene encodes P-glycoprotein implicated in the development of cellular drug resistance. The aim of this study was to develop high-resolution melting (HRM) analysis for determination of ABCB1 polymorphisms and evaluate their associations with clinical data of breast carcinoma patients. METHODS: HRM analysis was designed to assess five single nucleotide polymorphisms (SNPs) in ABCB1 (rs2214102, rs1128503, rs2032582, rs2032583 and rs1045642) in genomic DNA from 103 breast carcinoma patients. Results were confirmed by direct DNA sequencing. RESULTS: HRM analysis revealed distinct patterns of melting curves for the respective genotypes of all followed SNPs. Sensitivity of HRM analysis compared with direct DNA sequencing was superior (97.1% vs. 93.9%). The overall accuracy of HRM was 97.6%. The coefficients of variation in replicate experiments encompassed the range 0.002%-0.038%. On the basis of the examined SNPs, one strong haplotype block containing rs2032582 and rs1128503 SNPs was identified. Significant associations of rs2032582 SNP with tumor size, negative HER-2/neu status, and family history of breast carcinoma were found. Patients carrying the ancestral homozygous genotype (GG) in rs2214102 had significantly worse progression-free survival in comparison with carriers of the non-ancestral allele (A) in the adjuvant set (p=0.005). CONCLUSIONS: A rapid, accurate, low-cost and time-effective method for screening ABCB1 SNPs was developed. Significant associations of ABCB1 rs2032582 and rs2214102 SNPs with prognostic factors and survival of patients were found.

• MeSH
• denaturace nukleových kyselin MeSH
• jednonukleotidový polymorfismus * MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu diagnóza   genetika   terapie   MeSH
• P-glykoprotein genetika   MeSH
• prognóza MeSH
• sekvenční analýza DNA metody   MeSH
• tranzitní teplota * MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Doxorubicin belongs to anthracycline cytotoxic drugs and it is widely used as a major therapeutic agent in the treatment of various types of tumors. However,its therapeutic use is limited by the development of myelosuppression and cardiotoxicity after a specific cumulative dose is reached. The aim of this study was to investigate the effect of flavonoids, either natural or synthetic on doxorubicin-mediated formation of oxidative stress implicated in doxorubicin toxicity. Doxorubicin caused a concentration-dependent increase in the formation of hydroxyl radicals in minipig liver microsomes used as an in-vitro model system. When bacterial membranes heterologously expressing human NADPH cytochrome-P450 oxidoreductase were incubated with doxorubicin, formation of the superoxide radical under aerobic conditions and the doxorubicin–semiquinone radical under anaerobic conditions was detected. Forty different flavonoids were tested for their potency to prevent NADPH-induced or Fe2+-induced peroxidation of lipids in the microsomal system. According to the results, seven flavonoids were selected for evaluation of their potency to inhibit doxorubicin-dependent formation of hydroxyl radicals assessed by electron spin resonance. Myricetin, fisetin, and kaempferol were found to produce a significant protective effect against hydroxyl radicals in the minipig liver microsomal system. In conclusion, this study shows the use of a novel cost-effective in-vitro model system for preselection of antioxidants for testing of their protective effects against toxicity of anthracyclines and potentially other oxidative stress-inducing chemicals.

• MeSH
• antibiotika antitumorózní farmakologie   MeSH
• benzochinony metabolismus   MeSH
• doxorubicin farmakologie   toxicita   MeSH
• elektronová paramagnetická rezonance MeSH
• flavonoidy farmakologie   MeSH
• hydroxylový radikál metabolismus   MeSH
• jaterní mikrozomy účinky léků   metabolismus   MeSH
• lidé MeSH
• modely u zvířat MeSH
• NADPH-cytochrom c-reduktasa metabolismus   MeSH
• oxidační stres účinky léků   MeSH
• peroxidace lipidů účinky léků   MeSH
• prasata MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• superoxidy metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Checkpoint kinase 2 gene (CHEK2) alterations increase risk of several cancer types. We analyzed selected CHEK2 alterations in 270 Czech pancreatic cancer patients and in 683 healthy controls. The pancreatic cancer risk was higher in individuals who inherited rare alterations in CHEK2 region involving forkhead-associated domain other than I157T (OR=5.14; 95% CI=0.94-28.23) but the observed association was non-significant (p=0.057). The most frequent I157T mutation did not alter the pancreatic cancer risk and neither the followed deletion of 5395bp nor c.1100delC were found in any of pancreatic cases. We conclude that the I157T, other alterations in its proximity, del5395 and c.1100delC in CHEK2 do not predispose to pancreatic cancer risk in the Czech population.

• MeSH
• exony MeSH
• genetická predispozice k nemoci MeSH
• lidé MeSH
• nádory slinivky břišní enzymologie   epidemiologie   genetika   MeSH
• protein-serin-threoninkinasy genetika   MeSH
• sekvenční delece MeSH
• studie případů a kontrol MeSH
• terciární struktura proteinů MeSH
• tumor supresorové geny MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH