• Klíčová slova
• Mamma HELP,
• MeSH
• nádory prsu MeSH
• svépomocné skupiny MeSH
• Publikační typ
• biografie MeSH
• nekrology MeSH

• O autorovi
• Drexlerová, Jana, 1951-2020 Autorita

• Publikační typ
• novinové články MeSH
• rozhovory MeSH

BACKGROUND: Apoptosis plays a critical role in cancer cell survival and tumor development. We provide a hypothesis-generating screen for further research by exploring the expression profile and genetic variability of caspases (2, 3, 7, 8, 9, and 10) in breast carcinoma patients. This study addressed isoform-specific caspase transcript expression and genetic variability in regulatory sequences of caspases 2 and 9. METHODS: Gene expression profiling was performed by quantitative real-time PCR in tumor and paired non-malignant tissues of two independent groups of patients. Genetic variability was determined by high resolution melting, allelic discrimination, and sequencing analysis in tumor and peripheral blood lymphocyte DNA of the patients. RESULTS: CASP3 A+B and S isoforms were over-expressed in tumors of both patient groups. The CASP9 transcript was down-regulated in tumors of both groups of patients and significantly associated with expression of hormonal receptors and with the presence of rs4645978-rs2020903-rs4646034 haplotype in the CASP9 gene. Patients with a low intratumoral CASP9A/B isoform expression ratio (predicted to shift equilibrium towards anti-apoptotic isoform) subsequently treated with adjuvant chemotherapy had a significantly shorter disease-free survival than those with the high ratio (p=0.04). Inheritance of CC genotype of rs2020903 in CASP9 was associated with progesterone receptor expression in tumors (p=0.003). CONCLUSIONS: Genetic variability in CASP9 and expression of its splicing variants present targets for further study.

• MeSH
• cílená molekulární terapie * MeSH
• genetická transkripce * MeSH
• genetická variace * genetika   MeSH
• kaspasa 9 genetika   metabolismus   MeSH
• kaspasy * genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu * enzymologie   genetika   MeSH
• regulace genové exprese u nádorů * MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH

Hlavním záměrem projektu je validace a studium mechanismu účinku námi nedávno odhalených genetických markerů, tj. genové exprese v nádorových tkáních a genových polymorfismů v krvi pacientů léčených standardní, především neoadjuvantní chemoterapií karcinomu prsu. Cílem jsou markery metabolismu (např.: cytochromy P450, NQO, SOD, dráha 5-fluorouracilu) a transportu (ABC a SLC transportéry) cytostatik ve vztahu ke klinicko-patologickým charakteristikám, přežívání a účinnosti léčby pacientek. U úspěšně validovaných kandidátů budeme studovat mechanismus jakým markery působí korelací genové exprese s hladinou proteinu a zjištěním významu genotypu. Budeme studovat i methylaci regulačních oblastí kandidátních genů jako nezávislý marker. Zvláštní pozornost budeme věnovat studiu agresivních triple-negative karcinomů u pacientů i na modelových buněčných liniích s tímto fenotypem. U buněčných modelů se pokusíme vysledovat mechanismus nezávislou cestou včetně hledání vztahu mezi typem léčiva, jeho dávkou a účinkem; Validation of differences in expression of previously identified genetic markers between patients with breast carcinoma treated by standard, mainly neoadjuvant chemotherapy, will be the main goal of this project. Markers representing metabolism (e.g. cytochromes P450, SOD, NQO) and transport (ABC and SLC transporters) of anticancer drugs will be studied in relation to clinico-pathological characteristics, therapy outcome and survival of patients. Mechanism of action of validated markers will be studied by correlation of gene expression with protein level and analysis of genotype. Methylation of regulatory elements of candidate genes will be studied as independent prognostic or predictive marker. Special attention will be paid to the study of aggressive triple-negative tumors in both patients and cell models. Breast tumor cell line models differing in expression of receptors (estrogen, progesterone, ERBB2) will be employed for elucidation of mechanisms of action of biomarkers by alternative approach.

• MeSH
• ABC transportéry analýza   MeSH
• analýza přežití MeSH
• cytostatické látky MeSH
• exprese genu MeSH
• metylace MeSH
• nádorové biomarkery analýza   MeSH
• nádory prsu farmakoterapie   genetika   MeSH
• neoadjuvantní terapie MeSH
• polymorfismus genetický MeSH
• systém (enzymů) cytochromů P-450 analýza   MeSH
• triple-negativní karcinom prsu farmakoterapie   genetika   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• genetika, lékařská genetika
• biologie
• farmacie a farmakologie
• onkologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Microtubules are vitally important for eukaryotic cell division. Therefore, we evaluated the relevance of mitotic kinesin KIF14, protein-regulating cytokinesis 1 (PRC1), and citron kinase (CIT) for the prognosis of breast carcinoma patients. Transcript levels were assessed by quantitative real-time PCR in tissues from two independent groups of breast carcinoma patients and compared with clinical data. Tissue PRC1 protein levels were estimated using immunoblotting, and the PRC1 tagged haplotype was analyzed in genomic DNA. A functional study was performed in MDA-MB-231 cells in vitro. KIF14, PRC1, and CIT transcripts were overexpressed in tumors compared with control tissues. Tumors without expression of hormonal receptors or high-grade tumors expressed significantly higher KIF14 and PRC1 levels than hormonally-positive or low-grade tumors. Patients with high intra-tumoral PRC1 levels had significantly worse disease-free survival than patients with low levels. PRC1 rs10520699 and rs11852999 polymorphisms were associated with PRC1 transcript levels, but not with patientś survival. Paclitaxel-induced PRC1 expression, but PRC1 knockdown did not modify the paclitaxel cytotoxicity in vitro. PRC1 overexpression predicts poor disease-free survival of patients with breast carcinomas. Genetic variability of PRC1 and the protein interaction with paclitaxel cytotoxicity do not explain this association.

• MeSH
• buněčná smrt účinky léků   MeSH
• genový knockdown MeSH
• Kaplanův-Meierův odhad MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory prsu genetika   patologie   MeSH
• paclitaxel farmakologie   MeSH
• polymorfismus genetický MeSH
• přežití bez známek nemoci * MeSH
• proteiny buněčného cyklu genetika   metabolismus   MeSH
• regulace genové exprese u nádorů * účinky léků   MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

OBJECTIVES: ATP-Binding Cassette (ABC) transporters may cause treatment failure by transporting of anticancer drugs outside of the tumor cells. Multidrug resistance-associated protein 1 coded by the ABCC1 gene has recently been suggested as a potential prognostic marker in breast cancer patients. This study aimed to explore tagged haplotype covering nucleotide binding domain 1 of ABCC1 in relation with corresponding transcript levels in tissues and clinical phenotype of breast cancer patients. METHODS: The distribution of twelve ABCC1 polymorphisms was assessed by direct sequencing in peripheral blood DNA (n = 540). RESULTS: Tumors from carriers of the wild type genotype in rs35623 or rs35628 exhibited significantly lower levels of ABCC1 transcript than those from carriers of the minor allele (p = 0.003 and p = 0.004, respectively). The ABCC1 transcript levels significantly increased in the order CT-GT>CC-GT>CC-GG for the predicted rs35626-rs4148351 diplotype. Chemotherapy-treated patients carrying the T allele in rs4148353 had longer disease-free survival than those with the GG genotype (p = 0.043). On the other hand, hormonal therapy-treated patients with the AA genotype in rs35628 had significantly longer disease-free survival than carriers of the G allele (p = 0.012). CONCLUSIONS: Taken together, our study shows that genetic variability in the nucleotide binding domain 1 has a significant impact on the ABCC1 transcript level in the target tissue and may modify survival of breast cancer patients.

• MeSH
• analýza přežití * MeSH
• jednonukleotidový polymorfismus * MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• nádory prsu genetika   patofyziologie   MeSH
• proteiny spojené s mnohočetnou rezistencí k lékům genetika   metabolismus   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Metabolism of anticancer drugs affects their antitumor effects. This study has investigated the associations of gene expression of enzymes metabolizing anticancer drugs with therapy response and survival of breast carcinoma patients. Gene expression of 13 aldo-keto reductases (AKRs), carbonyl reductase 1, and 10 cytochromes P450 (CYPs) was assessed using quantitative real-time polymerase chain reaction in tumors and paired adjacent nonneoplastic tissues from 68 posttreatment breast carcinoma patients. Eleven candidate genes were then evaluated in an independent series of 50 pretreatment patients. Protein expression of the most significant genes was confirmed by immunoblotting. AKR1A1 was significantly overexpressed and AKR1C1-4, KCNAB1, CYP2C19, CYP3A4, and CYP3A5 downregulated in tumors compared with control nonneoplastic tissues after correction for multiple testing. Significant association of CYP2B6 transcript levels in tumors with expression of hormonal receptors was found in the posttreatment set and replicated in the pretreatment set of patients. Significantly higher intratumoral levels of AKR1C1, AKR1C2, or CYP2W1 were found in responders to neoadjuvant chemotherapy compared with nonresponders. Patients with high AKR7A3 or CYP2B6 levels in the pretreatment set had significantly longer disease-free survival than patients with low levels. Protein products of AKR1C1, AKR1C2, AKR7A3, CYP3A4, and carbonyl reductase (CBR1) were found in tumors and those of AKR1C1, AKR7A3, and CBR1 correlated with their transcript levels. Small interfering RNA-directed knockdown of AKR1C2 or vector-mediated upregulation of CYP3A4 in MDA-MB-231 model cell line had no effect on cell proliferation after paclitaxel treatment in vitro. Prognostic and predictive roles of drug-metabolizing enzymes strikingly differ between posttreatment and pretreatment breast carcinoma patients. Mechanisms of action of AKR1C2, AKR7A3, CYP2B6, CYP3A4, and CBR1 should continue to be further followed in breast carcinoma patients and models.

• MeSH
• aldehydreduktasa biosyntéza   genetika   MeSH
• imunoblotting MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• nádorové buňky kultivované MeSH
• nádory prsu genetika   metabolismus   patologie   MeSH
• prognóza MeSH
• proliferace buněk MeSH
• prospektivní studie MeSH
• průtoková cytometrie MeSH
• regulace genové exprese u nádorů * MeSH
• RNA nádorová genetika   MeSH
• systém (enzymů) cytochromů P-450 biosyntéza   genetika   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

AIM: ATP-binding cassette (ABC) transporters contribute to development of resistance to anticancer drugs via ATP-dependent drug efflux. A major goal of our study was to investigate associations between the expression of ABC transporters and outcome of breast carcinoma patients. PATIENTS & METHODS: Transcript levels of all 49 human ABC transporters were determined in post-treatment tumor and non-neoplastic tissue samples from 68 breast carcinoma patients treated by neoadjuvant chemotherapy. Six ABC transporters were then evaluated in independent series of 100 pretreatment patients. RESULTS: ABCA5/6/8/9/10, ABCB1/5/11, ABCC6/9, ABCD2/4, ABCG5 and ABCG8 were significantly downregulated and ABCA2/3/7/12, ABCB2/3/8/9/10, ABCC1/4/5/10/11/12, ABCD1/3, ABCE1, ABCF1/2/3 and ABCG1 were upregulated in post-treatment tumors compared with non-neoplastic tissues. Significant associations of intratumoral levels of ABCC1 and ABCC8 with grade and expression of hormonal receptors were found in both sets of patients. ABCA12, ABCA13 and ABCD2 levels were significantly associated with the response to neoadjuvant chemotherapy in post-treatment patients. Protein expression of ABCA12, ABCC8 and ABCD2 in tumor tissues of patients with breast carcinoma was observed by immunoblotting for the first time. CONCLUSION: ABCA12, ABCA13, ABCC1, ABCC8 and ABCD2 present potential modifiers of progression and response to the chemotherapy of breast carcinoma.

• MeSH
• ABC transportéry genetika   metabolismus   MeSH
• genetické asociační studie MeSH
• karcinom farmakoterapie   genetika   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu farmakoterapie   genetika   patologie   MeSH
• neoadjuvantní terapie MeSH
• protinádorové látky aplikace a dávkování   MeSH
• regulace genové exprese u nádorů MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

AIM: A role of caspase-2 in chemotherapy-induced apoptosis has been suggested. Our study aimed to evaluate the prognostic and predictive importance of caspase-2 isoforms in breast cancer patients. MATERIALS & METHODS: Caspase-2L and -2S transcript levels were determined in paired tumor and non-malignant control tissues from 64 patients after neoadjuvant chemotherapy and 100 pretreatment patients (general set) by real-time PCR with absolute quantification. RESULTS: Low but statistically significant upregulation of caspase-2L in tumor versus control tissues was observed in both sets. Significant associations of the levels of caspase-2L, -2S or S/L ratio with clinical prognostic factors were observed. However, none of these associations were confirmed in both sets. Levels of caspase-2 isoforms or the S/L ratio did not significantly associate with progression-free survival in the general set or with chemotherapy response in the neoadjuvant set. CONCLUSION: Our results suggest that the role of caspase-2 isoforms in the progression of breast cancer may considerably differ between pre- and post-chemotherapy patients.

• MeSH
• adjuvantní chemoterapie MeSH
• cysteinové endopeptidasy genetika   metabolismus   MeSH
• dospělí MeSH
• duktální karcinom prsu enzymologie   mortalita   MeSH
• frekvence genu MeSH
• izoenzymy genetika   metabolismus   MeSH
• Kaplanův-Meierův odhad MeSH
• kaspasa 2 genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• nádory prsu enzymologie   mortalita   MeSH
• neoadjuvantní terapie MeSH
• sekvenční analýza DNA MeSH
• senioři MeSH
• upregulace MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH