The development of colon cancer, one of the most common malignancies, is accompanied with numerous lipid alterations. However, analyses of whole tumor samples may not always provide an accurate description of specific changes occurring directly in tumor epithelial cells. Here, we analyzed in detail the phospholipid (PL), lysophospholipid (lysoPL), and fatty acid (FA) profiles of purified EpCAM+ cells, isolated from tumor and adjacent non-tumor tissues of colon cancer patients. We found that a number of FAs increased significantly in isolated tumor cells, which also included a number of long polyunsaturated FAs. Higher levels of FAs were associated with increased expression of FA synthesis genes, as well as with altered expression of enzymes involved in FA elongation and desaturation, including particularly fatty acid synthase, stearoyl-CoA desaturase, fatty acid desaturase 2 and ELOVL5 fatty acid elongase 5 We identified significant changes in ratios of specific lysoPLs and corresponding PLs. A number of lysophosphatidylcholine and lysophosphatidylethanolamine species, containing long-chain and very-long chain FAs, often with high numbers of double bonds, were significantly upregulated in tumor cells. Increased de novo synthesis of very long-chain FAs, or, altered uptake or incorporation of these FAs into specific lysoPLs in tumor cells, may thus contribute to reprogramming of cellular phospholipidome and membrane alterations observed in colon cancer.
- MeSH
- adenokarcinom enzymologie genetika metabolismus MeSH
- desaturasy mastných kyselin genetika metabolismus MeSH
- elongasy mastných kyselin genetika metabolismus MeSH
- epitelové buňky enzymologie metabolismus MeSH
- fosfolipidy metabolismus MeSH
- lidé MeSH
- lipidomika MeSH
- lipogeneze MeSH
- mastné kyseliny metabolismus MeSH
- metabolismus lipidů * MeSH
- nádory tračníku enzymologie genetika metabolismus MeSH
- regulace genové exprese u nádorů * MeSH
- senioři MeSH
- stearyl-CoA-desaturasa genetika metabolismus MeSH
- syntázy mastných kyselin genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- adhezní molekula epiteliálních buněk metabolismus MeSH
- dospělí MeSH
- galaktosyltransferasy genetika MeSH
- kolorektální nádory metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- sfingolipidy metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Identification of changes of phospholipid (PL) composition occurring during colorectal cancer (CRC) development may help us to better understand their roles in CRC cells. Here, we used LC-MS/MS-based PL profiling of cell lines derived from normal colon mucosa, or isolated at distinct stages of CRC development, in order to study alterations of PL species potentially linked with cell transformation. We found that a detailed evaluation of phosphatidylinositol (PI) and phosphatidylserine (PS) classes allowed us to cluster the studied epithelial cell lines according to their origin: i) cells originally derived from normal colon tissue (NCM460, FHC); ii) cell lines derived from colon adenoma or less advanced differentiating adenocarcinoma cells (AA/C1, HT-29); or, iii) cells obtained by in vitro transformation of adenoma cells and advanced colon adenocarcinoma cells (HCT-116, AA/C1/SB10, SW480, SW620). Although we tentatively identified several PS and PI species contributing to cell line clustering, full PI and PS profiles appeared to be a key to the successful cell line discrimination. In parallel, we compared PL composition of primary epithelial (EpCAM-positive) cells, isolated from tumor and adjacent non-tumor tissues of colon cancer patients, with PL profiles of cell lines derived from normal colon mucosa (NCM460) and from colon adenocarcinoma (HCT-116, SW480) cells, respectively. In general, higher total levels of all PL classes were observed in tumor cells. The overall PL profiles of the cell lines, when compared with the respective patient-derived cells, exhibited similarities. Nevertheless, there were also some notable differences in levels of individual PL species. This indicated that epithelial cell lines, derived either from normal colon tissue or from CRC cells, could be employed as models for functional lipidomic analyses of colon cells, albeit with some caution. The biological significance of the observed PL deregulation, or their potential links with specific CRC stages, deserve further investigation.
- MeSH
- analýza hlavních komponent MeSH
- epitelové buňky metabolismus patologie MeSH
- fosfolipidy metabolismus MeSH
- kolon patologie MeSH
- lidé MeSH
- lipidomika * MeSH
- nádorové buněčné linie MeSH
- nádory tračníku metabolismus patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Dietary carcinogens, such as benzo[a]pyrene (BaP), are suspected to contribute to colorectal cancer development. n-3 Polyunsaturated fatty acids (PUFAs) decrease colorectal cancer risk in individuals consuming diets rich in PUFAs. Here, we investigated the impact of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid on metabolism and genotoxicity of BaP in human cell models derived from the colon: HT-29 and HCT-116 cell lines. Both PUFAs reduced levels of excreted BaP metabolites, in particular BaP-tetrols and hydroxylated BaP metabolites, as well as formation of DNA adducts in HT-29 and HCT-116 cells. However, EPA appeared to be a more potent inhibitor of formation of some intracellular BaP metabolites, including BaP-7,8-dihydrodiol. EPA also reduced phosphorylation of histone H2AX (Ser139) in HT-29 cells, which indicated that it may reduce further forms of DNA damage, including DNA double strand breaks. Both PUFAs inhibited induction of CYP1 activity in colon cells determined as 7-ethoxyresorufin-O-deethylase (EROD); this was at least partly linked with inhibition of induction of CYP1A1, 1A2 and 1B1 mRNAs. The downregulation and/or inhibition of CYP1 enzymes by PUFAs could thus alter metabolism and reduce genotoxicity of BaP in human colon cells, which might contribute to known chemopreventive effects of PUFAs in colon epithelium.
- MeSH
- adukty DNA metabolismus MeSH
- antikarcinogenní látky farmakologie MeSH
- benzopyren škodlivé účinky metabolismus MeSH
- epitelové buňky účinky léků MeSH
- histony metabolismus MeSH
- kontrolní body fáze S buněčného cyklu účinky léků MeSH
- kyselina eikosapentaenová farmakologie MeSH
- kyseliny dokosahexaenové farmakologie MeSH
- lidé MeSH
- mutageny škodlivé účinky metabolismus MeSH
- nádorové buněčné linie MeSH
- poškození DNA účinky léků MeSH
- rodina 1 cytochromu P450 metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The development and progression of colorectal cancer (CRC), a major cause of cancer-related death in the western world, is accompanied with alterations of sphingolipid (SL) composition in colon tumors. A number of enzymes involved in the SL metabolism have been found to be deregulated in human colon tumors, in experimental rodent studies, and in human colon cancer cells in vitro. Therefore, the enzymatic pathways that modulate SL levels have received a significant attention, due to their possible contribution to CRC development, or as potential therapeutic targets. Many of these enzymes are associated with an increased sphingosine-1-phosphate/ceramide ratio, which is in turn linked with increased colon cancer cell survival, proliferation and cancer progression. Nevertheless, more attention should also be paid to the more complex SLs, including specific glycosphingolipids, such as lactosylceramides, which can be also deregulated during CRC development. In this review, we focus on the potential roles of individual SLs/SL metabolism enzymes in colon cancer, as well as on the pros and cons of employing the current in vitro models of colon cancer cells for lipidomic studies investigating the SL metabolism in CRC.
- MeSH
- alkalická ceramidasa genetika metabolismus MeSH
- ceramidy metabolismus MeSH
- fosfotransferasy s alkoholovou skupinou jako akceptorem genetika metabolismus MeSH
- kyselá ceramidasa genetika metabolismus MeSH
- laktosylceramidy metabolismus MeSH
- lidé MeSH
- lysofosfolipidy metabolismus MeSH
- metabolismus lipidů genetika MeSH
- modely nemocí na zvířatech MeSH
- nádorové buňky kultivované MeSH
- nádory tračníku enzymologie genetika patologie MeSH
- neutrální ceramidasa genetika metabolismus MeSH
- protoonkogenní proteiny c-akt genetika metabolismus MeSH
- regulace genové exprese u nádorů * MeSH
- sfingolipidy metabolismus MeSH
- sfingosin-N-acyltransferasa genetika metabolismus MeSH
- sfingosin analogy a deriváty metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Butyrate helps to maintain colon homeostasis and exhibits chemopreventive effects in colon epithelium. We examined the interactive effects of butyrate and benzo[a]pyrene (BaP), dietary carcinogen, in regulation of expression of a panel of phase I and II xenobiotic metabolizing enzymes (XMEs) in human colon cells. In human colon carcinoma HCT-116 and HT-29 cell lines, butyrate alone increased mRNA levels of some enzymes, such as N-acetyltransferases (in particular NAT2). In combination with BaP, butyrate potentiated induction of cytochrome P450 family 1 enzymes (CYP1A1), aldo-keto reductases (AKR1C1) or UDP-glucuronosyltransferases (UGT1A1). There were some notable differences between cell lines, as butyrate potentiated induction of NAD(P)H:quinone oxidoreductase 1 (NQO1) and UGT1A4 only in HCT-116 cells, and it even repressed AKR1C3 induction in HT-29 cells. Butyrate also promoted induction of CYP1, NQO1, NAT2, UGT1A1 or UGT1A4 in human colon Caco-2 cells, in a differentiation-dependent manner. Differentiated Caco-2 cells exhibited a higher inducibility of selected XME genes than undifferentiated cells. Butyrate increased induction of enzymatic activities of NATs, NQO1 and UGTs by BaP in HCT-116 and HT29 cells, whereas in differentiated Caco-2 cells it helped to increase only enzymatic activity of NQO1 and UGTs. Together, the present data suggest that butyrate may modulate expression/activities of several enzymes involved in metabolism of carcinogens in colon. In some cases (NAT2, UGT1 A1), this was linked to inhibition of histone deacetylases (HDAC), as confirmed by using HDAC inhibitor trichostatin A. These results may have implications for our understanding of the role of butyrate in regulation of XMEs and carcinogen metabolism in colon.
- MeSH
- benzopyren toxicita MeSH
- buněčné linie MeSH
- butyráty farmakologie MeSH
- epitelové buňky účinky léků metabolismus MeSH
- karcinogeny toxicita MeSH
- kolon cytologie MeSH
- lidé MeSH
- oxidoreduktasy genetika metabolismus MeSH
- transferasy genetika metabolismus MeSH
- xenobiotika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Východiska: Rozvoj nádorového onemocnění tlustého střeva je mimo jiné charakterizován abnormalitami v syntéze a metabolizmu lipidů, ovlivňujícími energetickou rovnováhu, strukturu a funkci biologických membrán, produkci specifických mediátorů a buněčné signálování. Změny buněčného lipidového profilu a metabolizmu (lipidomu) zásadně ovlivňují chování buněk i jejich odpověď na terapii. K lepšímu pochopení problematiky na buněčné a molekulární úrovni jsou využívány permanentní linie epiteliálních buněk tlustého střeva různého stupně nádorového rozvoje. Naše práce vycházela z předpokladu, že detailní analýza lipidomu různých buněčných linií tlustého střeva může odhalit takové rozdíly v množství a profilu specifických tříd/typů lipidů, které se mohou podílet na jejich rozdílné odpovědi k různým podnětům. Materiál a metody: Z buněk šesti lidských epiteliálních linií tlustého střeva odvozených z tkáně v různém stupni nádorového rozvoje byly izolovány lipidy a podrobeny LC/MS/MS (kapalinová chromatografie s tandemovou hmotnostní spektrometrií) analýze. Bylo stanoveno množství a hmotnostní profily všech tříd fosfolipidů (PL), lysofosfolipidů (lysoPL) a sfingolipidů. Tato data byla matematicky vyhodnocena (shluková a diskriminační analýza) s ohledem na vzájemné srovnání linií a na významně diskriminující typy lipidů. Výsledky: Výsledky shlukové analýzy seřadily buněčné linie dle stupně jejich nádorové transformace (normální buňky, adenom, karcinom, lymfatická metastáza). Výsledky diskriminačních analýz odhalily nejvíce rozlišující typy lipidů i odlišnosti v poměru PL: lysoPL. Ukázaly se zejména významné korelace zastoupení a profilu některých specifických tříd lysoPL a sfingolipidů se stupněm nádorové transformace buněk. Podobné přístupy jsou nyní aplikovány při srovnání lipidomu střevních epiteliálních buněk izolovaných z nádorové vs. nenádorové tkáně pacientů s nádory tlustého střeva. Závěr: Naše výsledky ukázaly, že a) vybrané buněčné linie jsou vhodným modelem pro lipidomické studie a mohou sloužit jako základ k navazujícímu klinickému výzkumu, b) analýza buněčného lipidomu by mohla přispět k rozlišení nádorových a nenádorových buněk také u klinických vzorků, u nichž by specifické typy lipidů mohly být využity jako biomarkery.
Backgrounds: Colon cancer development is often characterized by abnormalities in lipid synthesis and metabolism, which may influence energetic balance, structure and function of biological membranes, or production of specific mediators and cell signalling. The changes in lipid profile and metabolism (lipidome) may significantly affect cell behaviour and response to therapy. Permanent epithelial cell lines at various stages of cancer development are used for better understanding of this topic on cellular and molecular levels. In our study, we hypothesized that detailed analyses of colon cancer cell line lipidomes may help to identify major alterations in the amount and profile of specific lipid classes/species, which can contribute to their different response to various stimuli. Material and Methods: Cellular lipids were isolated from six human epithelial cell lines derived from tissues at various stages of tumour development. Liquid chromatography coupled with tandem mass spectometry analyses were performed in order to determine amount and mass profiles of all phospholipid (PL), lysophospholipid (lysoPL) and sphingolipid classes. The data was statistically evaluated (cluster and discrimination analyses) with respect to mutual comparison of cell lines and to significantly discriminating lipid types. Results: The results of cluster analysis arranged cell lines in order corresponding to their level of transformation (normal cells, adenoma, carcinoma and lymph node metastasis). The results of discrimination analyses revealed the most discriminating lipid types and distinction in PL: lysoPL ratios. Particularly, significant correlation of the amount and profiles of both specific lysoPL and sphingolipid classes with cell transformation level were observed. Similar approaches are now applied to compare lipidomes of colon epithelial cells isolated from tumour vs. non-tumour samples of colon cancer patients. Conclusion: Our results indicate that a) selected cancer cell lines are suitable model for lipidomic studies that can serve as a basis for subsequent clinical research, b) cellular lipidome analyses may help to discriminate tumour and non-tumour cells in clinical samples, where specific types of lipids could serve as biomarkers.
- MeSH
- adenom patologie metabolismus MeSH
- chromatografie kapalinová MeSH
- diskriminační analýza MeSH
- fosfolipidy metabolismus MeSH
- karcinom patologie metabolismus MeSH
- lidé MeSH
- lymfatické metastázy patologie metabolismus MeSH
- lysofosfolipidy metabolismus MeSH
- metabolismus lipidů * MeSH
- nádorová transformace buněk * MeSH
- nádorové biomarkery MeSH
- nádorové buněčné linie MeSH
- nádory tračníku * metabolismus patologie MeSH
- sfingolipidy metabolismus MeSH
- shluková analýza MeSH
- tandemová hmotnostní spektrometrie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
