BACKGROUND: Caspase-8 and caspase-9 (encoded by CASP8 and CASP9) are executive caspases of programmed cell death (apoptosis). Dysregulation of apoptosis plays an important role in cancer development, progression, and resistance to anticancer therapy. The goal of this work was to evaluate potential associations between polymorphisms in CASP8 and CASP9, previously linked to breast cancer risk, and the transcript levels of these genes (including their alternative anti-apoptotic variants) in tumor tissues and the clinical characteristics of the patients. MATERIAL AND METHODS: Sanger sequencing, high resolution melting (HRM) analysis, and allelic discrimination were used to identify polymorphisms in DNA samples isolated from tumor tissues and peripheral blood lymphocytes of 60 breast carcinoma patients. Total transcript levels of CASP8 and CASP9, and levels of alternative splicing variants CASP8L and CASP9B, were quantified by real-time PCR in tumor tissues. Clinically interesting associations were validated in DNA from lymphocytes of 615 breast carcinoma patients. RESULTS: A haplotype in CASP9 composed of three polymorphisms rs4645978-rs2020903-rs4646034 was significantly associated with CASP9 expression in tumors, with the expression of the progesterone receptor and ERBB2, and with the TNBC subtype of breast carcinoma in the validation study. The associations between the rs3834129 polymorphism in CASP8 and stage of disease, rs6435074 with grade, expression of estrogen receptor and ERBB2, and rs6723097 with ERBB2 expression have not yet been validated. However, rs6723097 was associated with disease-free survival in patients treated with hormonal therapy. CONCLUSION: This study reveals a previously unknown and presumably functional (in silico) association between a haplotype in CASP9 and molecular and clinical phenotypes of breast carcinoma. The potential clinical utility of this association for prognostication of breast carcinoma should be evaluated by independent studies.Key words: breast carcinoma - caspases - polymorphisms - functional - clinical - importanceThis work was supported by grant of the CU Grant Agency No. 1444313, and grant of the Internal Grant Agency of the Czech Ministry of Health No. 15-25618A.The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 3. 3. 2016Accepted: 26. 10. 2016.

• MeSH
• karcinom chemie   farmakoterapie   genetika   MeSH
• kaspasa 8 genetika   MeSH
• kaspasa 9 genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu chemie   farmakoterapie   genetika   MeSH
• polymorfismus genetický MeSH
• přežití po terapii bez příznaků nemoci MeSH
• receptor erbB-2 analýza   MeSH
• receptory pro estrogeny analýza   MeSH
• receptory progesteronu analýza   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ERBB2 protein, human MeSH Prohlížeč
• kaspasa 8 MeSH
• kaspasa 9 MeSH
• receptor erbB-2 MeSH
• receptory pro estrogeny MeSH
• receptory progesteronu MeSH

BACKGROUND: Dysregulation of the balance between cell growth and death in the colonic epithelium is associated with cancer promotion. Understanding how cell death in this self-renewing tissue is regulated and how it is influenced by interaction of specific dietary components, especially fat and fibre, could lead to improved treatment and prevention strategies for cancer. AIM OF THE STUDY: The effects of two types of polyunsaturated fatty acids (PUFAs)--arachidonic (AA, 20:4, n-6) or docosahexaenoic (DHA, 22:6, n-3)--on the response of human colon adenocarcinoma HT-29 cells to sodium butyrate (NaBt) were investigated. METHODS: The parameters reflecting cell proliferation and cell death were studied together with oxidative response, mitochondrial membrane potential (MMP) and changes of selected regulatory molecules associated with cell cycle (p27(Kip1) and p21(Cip1/WAF1)) and apoptosis (caspase-3, caspase-9, poly (ADP-ribose) polymerase--PARP, Bcl-2, Bax, Bak,Mcl-1). RESULTS: We demonstrated that pre-treatment with either AA or DHA attenuated cell cycle arrest caused by NaBt which is associated with modulation of p27(Kip1), but not p21(Cip1/WAF1) protein expression. On the other hand, PUFAs sensitised HT-29 cells to NaBt-induced apoptosis. An increased amount of floating cells and cells in the subG(0)/G(1) population was associated with increased reactive oxygen species production, lipid peroxidation, decrease of MMP, activation of caspase-3 and -9, PARP cleavage, and decrease in the expression of antiapoptotic Mcl-1 protein. The observed effects were modulated by the addition of a protein synthesis inhibitor, cycloheximide, and partially reversed by the antioxidant Trolox. CONCLUSIONS: PUFAs may have beneficial effects in the colon enhancing apoptosis induced by NaBt. Alteration of cell membrane lipid composition and potentiation of oxidative processes accompanied by changes in mitochondria followed by stimulation of apoptotic cascade components play a role in these effects.

• MeSH
• adenokarcinom metabolismus   MeSH
• apoptóza * účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• buňky HT-29 MeSH
• butyráty metabolismus   MeSH
• fluorescenční mikroskopie MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy účinky léků   metabolismus   MeSH
• kyselina arachidonová aplikace a dávkování   MeSH
• kyseliny dokosahexaenové aplikace a dávkování   MeSH
• lidé MeSH
• membránové potenciály účinky léků   MeSH
• nádory tračníku metabolismus   MeSH
• nenasycené mastné kyseliny metabolismus   MeSH
• peroxidace lipidů účinky léků   MeSH
• poly(ADP-ribosa)-polymerasy účinky léků   metabolismus   MeSH
• protoonkogenní proteiny účinky léků   metabolismus   MeSH
• průtoková cytometrie MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• butyráty MeSH
• CASP3 protein, human MeSH Prohlížeč
• CASP9 protein, human MeSH Prohlížeč
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy MeSH
• kyselina arachidonová MeSH
• kyseliny dokosahexaenové MeSH
• nenasycené mastné kyseliny MeSH
• poly(ADP-ribosa)-polymerasy MeSH
• protoonkogenní proteiny MeSH
• reaktivní formy kyslíku MeSH

BACKGROUND: The aim of the study was to contribute to our understanding of the mechanisms responsible for the resistance of breast cancer cells to taxanes. MATERIALS AND METHODS: Cell cycle characteristics, DNA fragmentation, p53 and p21(WAF1/CIP1) expression, caspase-3 and caspase-9 activity, cytochrome c release from mitochondria during cell death induction by the taxanes paclitaxel and docetaxel in highly-sensitive MDA-MB-435 and highly-resistant NCI-ADR-RES human breast cancer cells were compared. RESULTS: Approximately 300-fold higher concentrations of the taxanes were required to induce death in resistant NCI-ADR-RES cells than in sensitive MDA-MB-435 cells. Cell death induced by the taxanes in both sensitive and resistant cells was preceded by the accumulation of cells in the G2/M-phase. Neither cell type produced any DNA fragmentation (DNA ladder) typical of regular apoptosis. The p53 and the p21(WAF1/CIP1) levels did not change in sensitive or in resistant cells during cell death induction by the taxanes. The activity of the executioner caspase-3 increased significantly (2 to 2.5-fold) and, similarly, the activity of caspase-9 increased significantly (2 to 3.5-fold) in both cell types. However, cytochrome c was found to be released from mitochondria into the cytosol only in the resistant NCI-ADR-RES cells, but not in the sensitive MDA-MB-435 cells. CONCLUSION: The death induced by the taxanes in the studied breast cancer cells can be characterized as an apoptosis-like death, including caspase-3 and caspase-9 activation but not oligonucleosomal DNA fragmentation. However, the mechanisms of death induction by the taxanes in sensitive MDA-MB-435 cells and resistant NCI-ADR-RES cells differ. Cytochrome c is released from the mitochondria in resistant but not in sensitive cells.

• MeSH
• antitumorózní látky fytogenní farmakologie   MeSH
• buněčná smrt účinky léků   fyziologie   MeSH
• buňky - růstové procesy účinky léků   MeSH
• chemorezistence MeSH
• cytochromy c metabolismus   MeSH
• docetaxel MeSH
• fragmentace DNA účinky léků   MeSH
• inhibitor p21 cyklin-dependentní kinasy biosyntéza   MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy metabolismus   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádorový supresorový protein p53 biosyntéza   MeSH
• nádory prsu farmakoterapie   metabolismus   patologie   MeSH
• paclitaxel farmakologie   MeSH
• taxoidy farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antitumorózní látky fytogenní MeSH
• CASP3 protein, human MeSH Prohlížeč
• CASP9 protein, human MeSH Prohlížeč
• CDKN1A protein, human MeSH Prohlížeč
• cytochromy c MeSH
• docetaxel MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy MeSH
• nádorový supresorový protein p53 MeSH
• paclitaxel MeSH
• taxoidy MeSH
• TP53 protein, human MeSH Prohlížeč

Taxane and platinum-based chemotherapy regimens are standard treatment for advanced ovarian carcinoma. Expression levels of putative markers of taxane resistance in carcinoma tissues and paired peritoneal samples (n=55) and in 16 samples of ovaries without signs of carcinoma were compared with clinical data and the patients' time to progression. KIF14, PRC1, CIT and ABCC1 genes were significantly overexpressed in carcinomas when compared with normal ovarian tissues, while ABCB1 and CASP9 expression was decreased. Associations of protein expression of the proliferation marker Ki-67 with KIF14, PRC1, ABCB1 and CASP2 were found. Lastly, it was discovered that ABCB1 and CASP2 levels associated with FIGO stage and that the CIT level associated with the time to progression of ovarian carcinoma patients (P<0.0001). In conclusion, ABCB1, CASP2, KIF14, PRC1 and CIT genes seem to associate with surrogate markers of ovarian carcinoma progression and CIT gene associates with therapy outcome.

• Klíčová slova
• Clinical course, Cytokinesis, Gene expression, Ovarian carcinoma, Resistance, Taxane,
• MeSH
• ABC transportéry genetika   MeSH
• adenokarcinom diagnóza   farmakoterapie   genetika   MeSH
• antitumorózní látky terapeutické užití   MeSH
• chemorezistence genetika   MeSH
• genetické asociační studie MeSH
• intracelulární signální peptidy a proteiny genetika   MeSH
• kaspasy genetika   MeSH
• kineziny genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory vaječníků diagnóza   farmakoterapie   genetika   MeSH
• onkogenní proteiny genetika   MeSH
• ovarium metabolismus   MeSH
• peritoneum metabolismus   MeSH
• přemostěné cyklické sloučeniny terapeutické užití   MeSH
• progrese nemoci MeSH
• protein-serin-threoninkinasy genetika   MeSH
• proteiny buněčného cyklu genetika   MeSH
• taxoidy terapeutické užití   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ABC transportéry MeSH
• antitumorózní látky MeSH
• citron-kinase MeSH Prohlížeč
• intracelulární signální peptidy a proteiny MeSH
• kaspasy MeSH
• KIF14 protein, human MeSH Prohlížeč
• kineziny MeSH
• onkogenní proteiny MeSH
• PRC1 protein, human MeSH Prohlížeč
• přemostěné cyklické sloučeniny MeSH
• protein-serin-threoninkinasy MeSH
• proteiny buněčného cyklu MeSH
• taxane MeSH Prohlížeč
• taxoidy MeSH

We studied morphological changes of the nucleoli in HeLa cells treated with cisplatin and compared them with induction of markers of programmed cell death and TUNEL staining. We used different light microscopic nucleolar staining methods allowing us to visualize not only nucleolar proteins but also nucleolar RNA. Our results show predominantly compact, centrally localized nucleoli in intact control HeLa cells. In cisplatin-treated HeLa cells, we found an early onset of nucleolar segregation of proteins detected by argyrophilic nucleolar organizer regions and anti-nucleolar monoclonal antibody as well as an increased immunoreactivity for activated caspase-3 after 6 hours. Staining with Toluidine Blue and Methyl-green Pyronine revealed segregated nucleoli 12 hours after the treatment with cisplatin. TUNEL positivity in cisplatin-treated HeLa cells was accompanied by the aggregation of the argyrophilic proteins in the central portion of nucleus, disappearance of nucleolar RNA and shrinkage of the nucleus after 24 hours. Monitoring of the biochemical changes by immunoblotting revealed that activation of distinct caspases and degradation of their downstream protein substrates is executed in two phases. During an early apoptotic stage beginning 4.5 hours post treatment an activation of caspase-9 and caspase-3 was observed. This was accompanied by proteolytic cleavage of poly(ADP-ribose) polymerase-1 (PARP-1). The caspase-9 activation seems to be mediated by recruitment by the activating factor Apaf-1 because the increased accumulation of Apaf-1 and cytochrome C in cytosol preceded the generation of mature caspase-9 form. A second phase of apoptosis occurring between 10 and 15 hours post treatment was characterized by degradation of other nucleolar and nuclear proteins such as nuclear lamins, topoisomerase I and B23. In conclusion, remarkable segregation of nucleolar argyrophilic proteins, nucleolar RNA and a simultaneous activation of the cascade of caspases markedly preceded the TUNEL positivity in cisplatin-treated HeLa cells thereby substantiating the hypothesis that the nucleolus is a preferred target for caspase-3-dependent proteolysis in cisplatin-treated HeLa cells.

• MeSH
• aktivace enzymů MeSH
• apoptóza účinky léků   MeSH
• buněčné jadérko ultrastruktura   MeSH
• cisplatina farmakologie   MeSH
• frakcionace buněk MeSH
• HeLa buňky MeSH
• hydrolýza MeSH
• imunohistochemie MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy metabolismus   MeSH
• koncové značení zlomů DNA in situ MeSH
• lidé MeSH
• poly(ADP-ribosa)-polymerasy metabolismus   MeSH
• velikost buňky účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CASP3 protein, human MeSH Prohlížeč
• CASP9 protein, human MeSH Prohlížeč
• cisplatina MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy MeSH
• poly(ADP-ribosa)-polymerasy MeSH

As an extension of our recently published work (Mlejnek and Kuglík [2000] J. Cell. Biochem. 77:6-17), the role of caspases in N(6)-benzylaminopurine riboside (BAPR)-induced apotosis in HL-60 cells was evaluated in this study. Here, BAPR-induced apoptosis was accompanied by activation of caspase-3 and caspase-9. However, when these caspases were selectively inhibited, the progression of BAPR-induced apoptosis was not markedly affected. Besides that, activation of caspase-3 and caspase-9 was found to be rather late event in apoptotic process. These results suggested that other caspases might be critically implicated. Indeed, pan-specific caspase inhibitor, Z-VAD-FMK, completely prevented DNA cleavage and apoptotic bodies formation. However, Z-VAD-FMK failed to prevent cell death and it was incapable to fully counteract the main apoptotic hallmark-chromatin condensation. Finally, our data indicate that cellular decision between apoptosis and necrosis is made upon the availability of both caspase proteases and intracellular ATP.

• MeSH
• adenosin analogy a deriváty   farmakologie   MeSH
• apoptóza účinky léků   MeSH
• buněčná membrána účinky léků   enzymologie   MeSH
• buněčné jádro účinky léků   enzymologie   metabolismus   MeSH
• chloromethylketony aminokyselin farmakologie   MeSH
• chromatin metabolismus   MeSH
• HL-60 buňky cytologie   účinky léků   enzymologie   MeSH
• inhibitory cysteinových proteinas farmakologie   MeSH
• inhibitory kaspas * MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy fyziologie   MeSH
• lidé MeSH
• oligopeptidy farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adenosin MeSH
• benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone MeSH Prohlížeč
• benzyloxycarbonyl-leucyl-glutamyl-histidyl-aspartic acid fluoromethyl ketone MeSH Prohlížeč
• benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone MeSH Prohlížeč
• CASP3 protein, human MeSH Prohlížeč
• CASP9 protein, human MeSH Prohlížeč
• chloromethylketony aminokyselin MeSH
• chromatin MeSH
• inhibitory cysteinových proteinas MeSH
• inhibitory kaspas * MeSH
• kaspasa 3 MeSH
• kaspasa 9 MeSH
• kaspasy MeSH
• N(6)-benzyladenosine MeSH Prohlížeč
• oligopeptidy MeSH

PURPOSE: Tumour cells killing by cytotoxic therapies largely depends on triggering the intrinsic apoptosome-mediated caspase activation pathway but it had never been evaluated whether the expression of transcripts encoding the core components of apoptosome pathway is altered in non-small cell lung carcinoma (NSCLC). METHODS: We investigated the expression status of several apoptosome pathway-related transcripts including Apaf-1, procaspase-9, -3, -6, -7 and Smac in tumour and lung tissue samples from 65 surgically treated NSCLC patients and in 10 NSCLC cell lines with using real time RT-PCR. RESULTS: NSCLC tissues and cell lines showed significantly increased expression of procaspase-9, -3, -6 and Smac mRNAs as compared to the lungs and expression of these transcripts was simultaneously upregulated in a subset of NSCLCs belonging to different histopathological type, grade and stage categories. The expression of procaspase-7 mRNA in NSCLC tissues and cell lines and lungs was not significantly different. By contrast, the expression of Apaf-1 mRNA was frequently downregulated in the tumours as compared to matched lungs. Nevertheless, the examined NSCLC cell lines showed significantly higher expression of Apaf-1 mRNA than the lungs. The expression of Apaf-1, procaspase-9 and -6 mRNAs was higher in lung adenocarcinomas as compared to squamous cell lung carcinomas but the expression levels of the studied apoptosome pathway-related transcripts in the tumours were independent of tumour's grade and stage. CONCLUSIONS: The results of the present study suggest that there is a subgroup of NSCLCs, which may be intrinsically primed for apoptosis through upregulated expression of transcripts encoding the apoptosome pathway components.

• MeSH
• aktiny metabolismus   MeSH
• apoptóza * genetika   MeSH
• dospělí MeSH
• down regulace MeSH
• faktor 1 aktivující apoptotickou proteasu MeSH
• genetická transkripce * MeSH
• intracelulární signální peptidy a proteiny genetika   metabolismus   MeSH
• kaspasa 3 MeSH
• kaspasa 6 MeSH
• kaspasa 7 MeSH
• kaspasa 9 MeSH
• kaspasy genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• mitochondriální proteiny genetika   metabolismus   MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• nádory plic genetika   metabolismus   MeSH
• nemalobuněčný karcinom plic genetika   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prekurzory enzymů metabolismus   MeSH
• proteiny regulující apoptózu MeSH
• proteiny genetika   metabolismus   MeSH
• regulace genové exprese u nádorů MeSH
• senioři MeSH
• upregulace MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• APAF1 protein, human MeSH Prohlížeč
• CASP3 protein, human MeSH Prohlížeč
• CASP7 protein, human MeSH Prohlížeč
• CASP9 protein, human MeSH Prohlížeč
• DIABLO protein, human MeSH Prohlížeč
• faktor 1 aktivující apoptotickou proteasu MeSH
• intracelulární signální peptidy a proteiny MeSH
• kaspasa 3 MeSH
• kaspasa 6 MeSH
• kaspasa 7 MeSH
• kaspasa 9 MeSH
• kaspasy MeSH
• messenger RNA MeSH
• mitochondriální proteiny MeSH
• nádorové biomarkery MeSH
• prekurzory enzymů MeSH
• proteiny regulující apoptózu MeSH
• proteiny MeSH