The present study highlights the important association between lipid alterations and differentiation/apoptotic responses in human colon differentiating (FHC) and nondifferentiating (HCT-116) cell lines after their treatment with short-chain fatty acid sodium butyrate (NaBt), polyunsaturated fatty acids (PUFAs), and/or their combination. Our data from GC/MS and LC/MS/MS showed an effective incorporation and metabolization of the supplemented arachidonic acid (AA) or docosahexaenoic acid (DHA), resulting in an enhanced content of the respective PUFA in individual phospholipid (PL) classes and an altered composition of the whole cellular fatty acid spectrum in both FHC and HCT-116 cells. We provide novel evidence that NaBt combined with PUFAs additionally modulated AA and DHA cellular levels and caused their shift from triacylglycerol to PL fractions. NaBt increased, while AA, DHA and their combination with NaBt decreased endogenous fatty acid synthesis in FHC but not in HCT-116 cells. Fatty acid treatment also altered membrane lipid structure, augmented cytoplasmic lipid droplet accumulation, reactive oxygen species (ROS) production and dissipation of the mitochondrial membrane potential. All these parameters were significantly enhanced by combined NaBt/PUFA treatment, but only in FHC cells was this accompanied by highly increased apoptosis and suppressed differentiation. Moreover, the most significant changes of ROS production, differentiation and apoptosis among the parameters studied, the highest effects of combined NaBt/PUFA treatment and a lower sensitivity of HCT-116 cells were confirmed using two-way ANOVA. Our results demonstrate an important role of fatty acid-induced lipid alterations in the different apoptotic/differentiation response of colon cells with various carcinogenic potential.

• MeSH
• apoptóza účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• butyráty farmakologie   MeSH
• chromatografie kapalinová MeSH
• epitelové buňky účinky léků   metabolismus   MeSH
• fosfolipidy metabolismus   MeSH
• HCT116 buňky MeSH
• kolon cytologie   MeSH
• kyselina arachidonová farmakologie   MeSH
• kyseliny dokosahexaenové farmakologie   MeSH
• lidé MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• triglyceridy metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• butyráty MeSH
• fosfolipidy MeSH
• kyselina arachidonová MeSH
• kyseliny dokosahexaenové MeSH
• reaktivní formy kyslíku MeSH
• triglyceridy MeSH

The resistance of transformed epithelial cells to a detachment-induced apoptosis (anoikis) can significantly affect their susceptibility to anticancer therapy. We showed that detachment of both fetal (FHC) and adenocarcinoma (HT-29) human colon epithelial cells resulted in the activation of the pro-survival Akt pathway, and significant changes in integrin-linked kinase (ILK) and focal adhesive kinase (FAK) phosphorylation. We demonstrated a detachment-induced and PI3K/Akt-mediated resistance to apoptotic effects of TRAIL, which was not associated with any changes in the cell surface TRAIL death receptor levels. Instead, a modulation of downstream intracellular signaling events was suggested to be involved. Our results may have important implications for optimization of new strategies in treatment of cancers at different stages of development.

• MeSH
• adenokarcinom enzymologie   patologie   MeSH
• aktivace enzymů účinky léků   MeSH
• anoikis účinky léků   MeSH
• buněčná adheze účinky léků   MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• buňky HT-29 MeSH
• chemorezistence účinky léků   MeSH
• epitelové buňky účinky léků   enzymologie   patologie   MeSH
• fokální adhezní tyrosinkinasy metabolismus   MeSH
• fosfatidylinositol-3-kinasy metabolismus   MeSH
• fosforylace účinky léků   MeSH
• lidé MeSH
• nádory tračníku enzymologie   patologie   MeSH
• plod cytologie   MeSH
• protein TRAIL farmakologie   MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• protoonkogenní proteiny c-akt metabolismus   MeSH
• receptory domény smrti metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fokální adhezní tyrosinkinasy MeSH
• integrin-linked kinase MeSH Prohlížeč
• protein TRAIL MeSH
• protein-serin-threoninkinasy MeSH
• protoonkogenní proteiny c-akt MeSH
• receptory domény smrti MeSH

TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) can selectively trigger apoptosis in various cancer cell types. However, many cancer cells are resistant to death receptor-mediated apoptosis. Combination therapy with platinum complexes may affect TRAIL-induced signaling via modulation of various steps in apoptotic pathways. Here, we show that cisplatin or a more potent platinum(IV) complex LA-12 used in 20-fold lower concentration enhanced killing effects of TRAIL in human colon and prostate cancer cell lines via stimulation of caspase activity and overall apoptosis. Both platinum complexes increased DR5 surface expression in colon cancer cells. Small interfering RNA-mediated DR5 silencing rescued cells from sensitizing effects of platinum drugs on TRAIL-induced caspase-8 activation and apoptosis, showing the functional importance of DR5 in the effects observed. In addition, both cisplatin and LA-12 triggered the relocalization of DR4 and DR5 receptors to lipid rafts and accelerated internalization of TRAIL, which may also affect TRAIL signaling. Collectively, modulations of the initial steps of the extrinsic apoptotic pathway at the level of DR5 and plasma membrane are important for sensitization of colon and prostate cancer cells to TRAIL-induced apoptosis mediated by LA-12 and cisplatin.

• MeSH
• amantadin analogy a deriváty   farmakologie   MeSH
• apoptóza účinky léků   fyziologie   MeSH
• cisplatina farmakologie   MeSH
• fluorescenční protilátková technika MeSH
• konfokální mikroskopie MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory metabolismus   MeSH
• organoplatinové sloučeniny farmakologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein TRAIL metabolismus   MeSH
• průtoková cytometrie MeSH
• RNA interference MeSH
• separace buněk MeSH
• signální transdukce účinky léků   fyziologie   MeSH
• TRAIL receptory metabolismus   MeSH
• transport proteinů účinky léků   MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• amantadin MeSH
• bis(acetato)(1-adamantylamine)amminedichloroplatinum(IV) MeSH Prohlížeč
• cisplatina MeSH
• organoplatinové sloučeniny MeSH
• protein TRAIL MeSH
• TRAIL receptory MeSH

BACKGROUND: LA-12 is a new platinum (IV) drug with promising cytotoxic effects in a wide range of cancer cell lines. Its confluence-dependent effects were compared with cisplatin (CDDP) and oxaliplatin (L-OHP) in HT-29 cells. MATERIALS AND METHODS: Cytotoxicity was determined by MTT test, eosin exclusion assay, and cell number quantification. The cell cycle was analysed using propidium iodide DNA staining (flow cytometry), apoptosis by phosphatidylserine externalisation (annexin-V assay), mitochondrial membrane potential by flow cytometry, nuclear morphology by means of fluorescence microscopy, and PARP cleavage by Western blotting. RESULTS: While L-OHP and CDDP were practically inactive in the subconfluent cell population, LA-12 showed a similar toxicity in both subconfluent and growing populations. All compounds induced apoptosis, although with different potentials. CONCLUSION: LA-12 was able to overcome confluence-dependent resistance of HT-29 cells observed for other platinum compounds, which may have potential therapeutic use in slowly growing tumours.

• MeSH
• adenokarcinom farmakoterapie   MeSH
• amantadin analogy a deriváty   farmakologie   MeSH
• apoptóza účinky léků   MeSH
• buněčná adheze účinky léků   MeSH
• buňky HT-29 MeSH
• chemorezistence MeSH
• cisplatina farmakologie   MeSH
• lidé MeSH
• nádory tračníku farmakoterapie   patologie   MeSH
• organoplatinové sloučeniny farmakologie   MeSH
• oxaliplatin MeSH
• protinádorové látky farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• amantadin MeSH
• bis(acetato)(1-adamantylamine)amminedichloroplatinum(IV) MeSH Prohlížeč
• cisplatina MeSH
• organoplatinové sloučeniny MeSH
• oxaliplatin MeSH
• protinádorové látky MeSH

In addition to its ability to act as a promising inducer of tumor-specific cell death, TRAIL has also been shown to stimulate signaling pathways leading to cancer cell survival. We examined the changes of anti-apoptotic Mcl-1 protein level following TRAIL treatment of human cell lines representing different stages of colon carcinogenesis-adenocarcinoma (HT-29, HCT116) or secondary metastasis (SW620), together with cell line derived from human fetal colon (FHC). While TRAIL was capable of triggering an anti-apoptotic signaling leading to significant early ERK-mediated transcriptional up-regulation of Mcl-1 in selected colon adenocarcinoma cell lines, none or very limited effects were demonstrated in cell lines derived from colon lymph node metastasis or fetal colon, respectively. We demonstrated an immediate impact of Mcl-1 protein level manipulations on the course of early acute apoptotic response of colon adenocarcinoma cells to TRAIL. It is therefore essential to consider the dynamics of modulation of Mcl-1 level and the balance between TRAIL-induced pro- and anti-apoptotic pathways when predicting the response of cells in different stages of cancer development, and designing the anticancer therapy using TRAIL.

• MeSH
• epitelové buňky metabolismus   MeSH
• lidé MeSH
• metastázy nádorů patologie   MeSH
• nádorové buněčné linie MeSH
• nádory tračníku metabolismus   patologie   MeSH
• protein MCL-1 MeSH
• protein TRAIL fyziologie   MeSH
• protoonkogenní proteiny c-bcl-2 genetika   MeSH
• regulace genové exprese MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• protein MCL-1 MeSH
• protein TRAIL MeSH
• protoonkogenní proteiny c-bcl-2 MeSH

We verified the hypothesis suggesting modulation of the effects of sodium butyrate (NaBt) by omega-3 or omega-6 PUFAs. Comparing the response of human colon epithelial cell lines of fetal (FHC) and adenocarcinoma (HT-29, HCT-116) origin, we detected significant differences in proliferation, differentiation and apoptotic response to the treatment of NaBt, arachidonic or docosahexaenoic acids and their combination. While in FHC and HT-29 cells NaBt induced G0/G1 arrest, differentiation and low level of apoptosis, in HCT-116 cells G2/M arrest, no differentiation and high degree of apoptosis were detected. Moreover, in FHC cells significant potentiation of apoptosis accompanied by increased arrest in the cell cycle, cell detachment and decrease in differentiation were detected after combined treatment with NaBt and both PUFAs. Changes in cytokinetics induced by fatty acids were accompanied by membrane lipid unpacking, reactive oxygen species (ROS) production, and decrease in mitochondrial membrane potential (MMP). Detection of caspase-3 activation and dynamic modulation of Mcl-1 protein expression imply their possible role in both cell differentiation and apoptotic response. Our results support the concept of modulation of NaBt effects by PUFAs, especially of omega-3 type, in colonic cells in vitro with diverse impact in cell lines derived from normal or neoplastic epithelium.

• MeSH
• antioxidancia farmakologie   MeSH
• apoptóza účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčné dělení účinky léků   MeSH
• buněčné linie MeSH
• buňky HT-29 MeSH
• butyráty farmakologie   MeSH
• epitelové buňky účinky léků   MeSH
• HCT116 buňky MeSH
• kolon účinky léků   embryologie   MeSH
• lidé MeSH
• membránové lipidy chemie   MeSH
• nádory tračníku patologie   MeSH
• nenasycené mastné kyseliny farmakologie   MeSH
• plod MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• antioxidancia MeSH
• butyráty MeSH
• membránové lipidy MeSH
• nenasycené mastné kyseliny MeSH

The impressive impact of cisplatin on cancer on one side and severe side effects, as well as the development of drug resistance during treatment on the other side, were the factors motivating scientists to design and synthesize new more potent analogues lacking disadvantages of cisplatin. Platinum(IV) complexes represent one of the perspective groups of platinum-based drugs. In this review, we summarize recent findings on both in vitro and in vivo effects of platinum(IV) complexes with adamantylamine. Based on a literary overview of the mechanisms of activity of platinum-based cytostatics, we discuss opportunities for modulating the effects of novel platinum complexes through interactions with apoptotic signaling pathways and with cellular lipids, including modulations of the mitochondrial cell death pathway, oxidative stress, signaling of death ligands, lipid metabolism/signaling, or intercellular communication. These approaches might significantly enhance the efficacy of both novel and established platinum-based cytostatics.

• Publikační typ
• časopisecké články MeSH

We compared the response of normal (FHC) and cancer (HT-29) human colon epithelial cells to the important apoptotic inducers TNF-alpha, anti-Fas antibody and TNF-related apoptosis inducing ligand (TRAIL). The two cell lines did not respond to TNF-alpha (15 ng/ml), expressed a limited sensitivity to anti-Fas antibody (200 ng/ml) and a different response to TRAIL (100 ng/ml). We studied apoptosis with regard to the changes at the receptor level (DR, DcR and FLIP) and at the level of mitochondria (Bid protein cleavage, Apo2.7 protein expression and caspase-9 activation). Two different approaches were used to sensitize the cells to TRAIL-induced apoptosis: inhibition of protein synthesis (cycloheximide, CHX) and inhibition of the pro-survival MEK/ERK pathway (U0126). While the two cell lines were markedly sensitized to all three TNF family members by CHX, a different degree of response (especially for TRAIL) was obtained when inhibition of the MEK/ERK pathway was achieved. TRAIL-induced apoptosis was significantly enhanced by U0126 co-treatment in the HT-29 cells, but not in the FHC cells. The most significant differences between the HT-29 and FHC cells co-treated with TRAIL and U0126 were demonstrated with regard to the involvement of the mitochondrial apoptotic pathway, suggesting its importance in the regulation of cell sensitivity to the TRAIL-induced apoptosis.

• MeSH
• apoptóza MeSH
• buněčné linie MeSH
• epitelové buňky účinky léků   metabolismus   MeSH
• karcinom metabolismus   patologie   MeSH
• kaspasy metabolismus   MeSH
• lidé MeSH
• mitochondriální proteiny metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory tračníku metabolismus   patologie   MeSH
• proliferace buněk účinky léků   MeSH
• protein TRAIL farmakologie   MeSH
• protilátky farmakologie   MeSH
• protoonkogenní proteiny c-bcl-2 metabolismus   MeSH
• TNF decoy receptory metabolismus   MeSH
• TNF-alfa farmakologie   MeSH
• TRAIL receptory metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CH-11 anti-fas antibody, human MeSH Prohlížeč
• kaspasy MeSH
• mitochondriální proteiny MeSH
• protein TRAIL MeSH
• protilátky MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• TNF decoy receptory MeSH
• TNF-alfa MeSH
• TNFRSF10D protein, human MeSH Prohlížeč
• TRAIL receptory MeSH

Epithelial cells can be manipulated to undergo apoptosis depending on the balance between pro-survival and apoptotic signals. We showed that TRAIL-induced apoptosis may be differentially regulated by inhibitors of MEK ERK (U0126) or PI3K/Akt (LY294002) pathway in TRAIL-sensitive (HT-29) and TRAIL-resistant (SW620) human epithelial colon cancer cells. U0126 or LY294002 significantly enhanced TRAIL-induced apoptosis in HT-29 cells, but not in SW620 cells. We report a different regulation of the level of an anti-apoptotic Mcl-1 protein under MEK/ERK or PI3K/Akt pathway inhibition and suggest the mechanisms involved. A special attention was paid to the role of the ERK1/2, Akt, and glycogen synthase kinase 3beta.

• MeSH
• aktivace enzymů účinky léků   MeSH
• apoptóza účinky léků   MeSH
• buňky HT-29 MeSH
• extracelulárním signálem regulované MAP kinasy antagonisté a inhibitory   MeSH
• fosforylace účinky léků   MeSH
• inhibitory enzymů farmakologie   MeSH
• inhibitory fosfoinositid-3-kinasy MeSH
• kaspasa 8 metabolismus   MeSH
• keratin-18 metabolismus   MeSH
• kinasa 3 glykogensynthasy metabolismus   MeSH
• kinasa glykogensynthasy 3beta MeSH
• lidé MeSH
• mitogenem aktivovaná proteinkinasa 1 antagonisté a inhibitory   MeSH
• mitogenem aktivovaná proteinkinasa 3 antagonisté a inhibitory   MeSH
• mitogenem aktivované proteinkinasy kinas antagonisté a inhibitory   MeSH
• nádorové proteiny metabolismus   MeSH
• nádory tračníku patologie   MeSH
• poly(ADP-ribosa)polymerasy metabolismus   MeSH
• protein MCL-1 MeSH
• protein TRAIL farmakologie   MeSH
• protoonkogenní proteiny c-akt antagonisté a inhibitory   MeSH
• protoonkogenní proteiny c-bcl-2 metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• extracelulárním signálem regulované MAP kinasy MeSH
• GSK3B protein, human MeSH Prohlížeč
• inhibitory enzymů MeSH
• inhibitory fosfoinositid-3-kinasy MeSH
• kaspasa 8 MeSH
• keratin-18 MeSH
• kinasa 3 glykogensynthasy MeSH
• kinasa glykogensynthasy 3beta MeSH
• mitogenem aktivovaná proteinkinasa 1 MeSH
• mitogenem aktivovaná proteinkinasa 3 MeSH
• mitogenem aktivované proteinkinasy kinas MeSH
• nádorové proteiny MeSH
• poly(ADP-ribosa)polymerasy MeSH
• protein MCL-1 MeSH
• protein TRAIL MeSH
• protoonkogenní proteiny c-akt MeSH
• protoonkogenní proteiny c-bcl-2 MeSH

The resistance of some cancer cells to TRAIL-induced apoptosis is a major obstacle in successful clinical application of this cytokine. Combination treatment with agents capable of sensitising the cells to TRAIL effects is beneficial for new cancer treatment strategies. Docosahexaenoic acid (DHA) is under intense investigation for its ability to affect cancer cell growth and apoptosis. We demonstrated a modulation of TRAIL-induced apoptosis of HT-29 human colon cancer cells by DHA on the molecular (pro-caspase-3, -8, Bid, PARP cleavage) and cellular (cell viability and adhesion) level. To conclude, TRAIL and DHA were shown to cooperate in the induction of colon cancer cell apoptosis.

• MeSH
• adenokarcinom patologie   MeSH
• apoptóza fyziologie   MeSH
• buněčná adheze MeSH
• kyseliny dokosahexaenové farmakologie   MeSH
• lékové interakce MeSH
• lidé MeSH
• membránové glykoproteiny fyziologie   MeSH
• nádorové buňky kultivované MeSH
• nádory tračníku patologie   MeSH
• protein TRAIL MeSH
• proteiny regulující apoptózu MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• TNF-alfa fyziologie   MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kyseliny dokosahexaenové MeSH
• membránové glykoproteiny MeSH
• protein TRAIL MeSH
• proteiny regulující apoptózu MeSH
• TNF-alfa MeSH
• TNFSF10 protein, human MeSH Prohlížeč