Naturally-occurring mixtures of phytochemicals present in plant foods are proposed to possess tumor-suppressive activities. In this work, we aimed to evaluate the antitumor effects of Thymus vulgaris L. in in vivo and in vitro mammary carcinoma models. Dried T. vulgaris (as haulm) was continuously administered at two concentrations of 0.1% and 1% in the diet in a chemically-induced rat mammary carcinomas model and a syngeneic 4T1 mouse model. After autopsy, histopathological and molecular analyses of rodent mammary carcinomas were performed. In addition, in vitro evaluations using MCF-7 and MDA-MB-231 cells were carried out. In mice, T. vulgaris at both doses reduced the volume of 4T1 tumors by 85% (0.1%) and 84% (1%) compared to the control, respectively. Moreover, treated tumors showed a substantial decrease in necrosis/tumor area ratio and mitotic activity index. In the rat model, T. vulgaris (1%) decreased the tumor frequency by 53% compared to the control. Analysis of the mechanisms of anticancer action included well-described and validated diagnostic and prognostic markers that are used in both clinical approach and preclinical research. In this regard, the analyses of treated rat carcinoma cells showed a CD44 and ALDH1A1 expression decrease and Bax expression increase. Malondialdehyde (MDA) levels and VEGFR-2 expression were decreased in rat carcinomas in both the T. vulgaris treated groups. Regarding the evaluations of epigenetic changes in rat tumors, we found a decrease in the lysine methylation status of H3K4me3 in both treated groups (H3K9m3, H4K20m3, and H4K16ac were not changed); up-regulations of miR22, miR34a, and miR210 expressions (only at higher doses); and significant reductions in the methylation status of four gene promoters-ATM serin/threonine kinase, also known as the NPAT gene (ATM); Ras-association domain family 1, isoform A (RASSF1); phosphatase and tensin homolog (PTEN); and tissue inhibitor of metalloproteinase-3 (TIMP3) (the paired-like homeodomain transcription factor (PITX2) promoter was not changed). In vitro study revealed the antiproliferative and proapoptotic effects of essential oils of T. vulgaris in MCF-7 and MDA-MB-231 cells (analyses of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS); 5-bromo-20-deoxyuridine (BrdU); cell cycle; annexin V/PI; caspase-3/7; Bcl-2; PARP; and mitochondrial membrane potential). T. vulgaris L. demonstrated significant chemopreventive and therapeutic activities against experimental breast carcinoma.

• Klíčová slova
• MCF-7 cells, MDA-MB-231 cells, Thymus vulgaris, angiogenesis, apoptosis, cancer stem cells, cell proliferation, epigenetics, mammary carcinogenesis, predictive and preventive medicine, rat,
• MeSH
• epigeneze genetická účinky léků   MeSH
• fytoterapie MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory prsu farmakoterapie   MeSH
• oleje prchavé aplikace a dávkování   farmakologie   MeSH
• oleje rostlin aplikace a dávkování   farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• Thymus (rostlina) chemie   MeSH
• viabilita buněk MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• xenogenní modely - testy protinádorové aktivity MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• oleje prchavé MeSH
• oleje rostlin MeSH

It is supposed that plant functional foods, rich in phytochemicals, may potentially have preventive effects in carcinogenesis. In this study, the anticancer effects of cloves in the in vivo and in vitro mammary carcinoma model were assessed. Dried flower buds of cloves (CLOs) were used at two concentrations of 0.1% and 1% through diet during 13 weeks after the application of chemocarcinogen. After autopsy, histopathological and immunohistochemical analyses of rat mammary carcinomas were performed. Moreover, in vitro evaluation using MCF-7 cells was carried out. Dietary administered CLO caused the dose-dependent decrease in tumour frequency by 47.5% and 58.5% when compared to control. Analysis of carcinoma cells in animals showed bcl-2, Ki67, VEGFA, CD24 and CD44 expression decrease and Bax, caspase-3 and ALDH1 expression increase after high-dose CLO administration. MDA levels were substantially decreased in rat carcinomas in both CLO groups. The evaluation of histone modifications revealed increase in lysine trimethylations and acetylations (H4K20me3, H4K16ac) in carcinomas after CLO administration. TIMP3 promoter methylation levels of CpG3, CpG4, CpG5 islands were altered in treated cancer cells. An increase in total RASSF1A promoter methylation (three CpG sites) in CLO 1 group was found. In vitro studies showed antiproliferative and pro-apoptotic effects of CLO extract in MCF-7 cells (analyses of cytotoxicity, Brdu, cell cycle, annexin V/PI, caspase-7, Bcl-2 and mitochondrial membrane potential). This study showed a significant anticancer effect of clove buds in the mammary carcinoma model in vivo and in vitro.

• Klíčová slova
• MCF-7 cells, angiogenesis, apoptosis, cancer stem cells, cell proliferation, cloves, epigenetics, mammary carcinogenesis, rat,
• MeSH
• adenokarcinom dietoterapie   genetika   metabolismus   patologie   MeSH
• antigen Ki-67 genetika   metabolismus   MeSH
• epigeneze genetická účinky léků   MeSH
• experimentální nádory mléčných žláz dietoterapie   genetika   metabolismus   patologie   MeSH
• fytogenní protinádorové látky izolace a purifikace   farmakologie   MeSH
• histony genetika   metabolismus   MeSH
• kaspasa 3 genetika   metabolismus   MeSH
• krysa rodu Rattus MeSH
• květy chemie   MeSH
• lidé MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• metylace DNA účinky léků   MeSH
• MFC-7 buňky MeSH
• nádorové supresorové proteiny genetika   metabolismus   MeSH
• nádory prsu dietoterapie   genetika   metabolismus   patologie   MeSH
• potkani Sprague-Dawley MeSH
• protein X asociovaný s bcl-2 genetika   metabolismus   MeSH
• protoonkogenní proteiny c-bcl-2 genetika   metabolismus   MeSH
• retinaldehydrogenasa genetika   metabolismus   MeSH
• rodina enzymů aldehyddehydrogenasy 1 MeSH
• rostlinné extrakty chemie   MeSH
• signální transdukce MeSH
• Syzygium chemie   MeSH
• tumor burden účinky léků   MeSH
• vaskulární endoteliální růstový faktor A genetika   metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Aldh1a1 protein, rat MeSH Prohlížeč
• antigen Ki-67 MeSH
• Bax protein, rat MeSH Prohlížeč
• Bcl2 protein, rat MeSH Prohlížeč
• Casp3 protein, rat MeSH Prohlížeč
• fytogenní protinádorové látky MeSH
• histony MeSH
• kaspasa 3 MeSH
• nádorové supresorové proteiny MeSH
• protein X asociovaný s bcl-2 MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• RASSF1 protein, rat MeSH Prohlížeč
• retinaldehydrogenasa MeSH
• rodina enzymů aldehyddehydrogenasy 1 MeSH
• rostlinné extrakty MeSH
• vascular endothelial growth factor A, rat MeSH Prohlížeč
• vaskulární endoteliální růstový faktor A MeSH

PURPOSE: There has been a considerable interest in the identification of natural plant foods for developing effective agents against cancer. Thus, the anti-tumour effects of oregano in the in vivo and in vitro breast cancer model were evaluated. METHODS: Lyophilized oregano (ORE) was administered at two concentrations of 0.3 and 3 % through diet. The experiment was terminated 14 weeks after carcinogen administration. At autopsy, mammary tumours were removed and prepared for histopathological and immunohistochemical analysis. Moreover, in vitro evaluation in MCF-7 cells was carried out. RESULTS: Low-dose ORE suppressed tumour frequency by 55.5 %, tumour incidence by 44 %, and tumour volume by 44.5 % compared to control animals. Analysis of rat tumour cells showed Ki67, VEGFR-2, CD24, and EpCAM expression decrease and caspase-3 expression increase after low-dose ORE treatment. High-dose ORE lengthened tumour latency by 12.5 days; moreover, Bcl-2, VEGFR-2, CD24, and EpCAM expression decrease and caspase-3 expression increase in carcinoma cells were observed. Histopathological analysis revealed a decrease in the ratio of high-/low-grade carcinomas in both treated groups. In vitro studies showed that ORE decreased survival and proliferation of MCF-7 cells. In ORE-treated MCF-7 cells, an increase in cells expressing sub-G 0/G 1 DNA content and an increase in the percentage of annexin V/PI positive MCF-7 cells were observed. In vitro, both caspase-dependent and possible non-caspase-dependent apoptotic pathways were found. The deactivation of anti-apoptotic activity of Bcl-2, a decrease in mitochondrial membrane potential, and the activation of mitochondrial apoptosis pathway were observed in the ORE-treated MCF-7 cells. CONCLUSIONS: Our results demonstrate, for the first time, a distinct tumour-suppressive effect of oregano in the breast cancer model.

• Klíčová slova
• Angiogenesis, Apoptosis, Cancer stem cells, Cell proliferation, MCF-7 cells, Mammary carcinogenesis, Oregano, Rat,
• MeSH
• apoptóza účinky léků   MeSH
• dobromysl (rod) chemie   MeSH
• fytoterapie * MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• lyofilizace MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• MFC-7 buňky MeSH
• mitochondrie účinky léků   metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• nádory prsu farmakoterapie   MeSH
• potkani Sprague-Dawley MeSH
• proliferace buněk účinky léků   MeSH
• rostlinné přípravky farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• rostlinné přípravky MeSH

PURPOSE: Fruit and vegetable intake is inversely correlated with cancer; thus, it is proposed that an extract of phytochemicals as present in whole fruits, vegetables, or grains may have anti-carcinogenic properties. Thus, the anti-tumour effects of fruit peel polyphenols (Flavin7) in the chemoprevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis in female rats were evaluated. METHODS: Lyophilized substance of Flavin7 (F7) was administered at two concentrations of 0.3 and 3 % through diet. The experiment was terminated 14 weeks after carcinogen administration, and mammary tumours were removed and prepared for histopathological and immunohistochemical analysis. In addition, using an in vitro cytotoxicity assay, apoptosis and proliferation after F7 treatment in human breast adenocarcinoma (MCF-7) cells were performed. RESULTS: High-dose F7 suppressed tumour frequency by 58 % (P < 0.001), tumour incidence by 24 % (P < 0.05), and lengthened latency by 8 days (P > 0.05) in comparison with the control rats, whereas lower dose of F7 was less effective. Histopathological analysis of tumours showed significant decrease in the ratio of high-/low-grade carcinomas after high-dose F7 treatment. Immunohistochemical analysis of rat carcinoma cells in vivo found a significant increase in caspase-3 expression and significant decrease in Bcl-2, Ki67, and VEGFR-2 expression in the high-dose group. Both doses demonstrated significant positive effects on plasma lipid metabolism in rats. F7 significantly decreased survival of MCF-7 cells in vitro in MTT assay by dose- and time-dependent manner compared to control. F7 prevented cell cycle progression by significant enrichment in G1 cell populations. Incubation with F7 showed significant increase in the percentage of annexin V-/PI-positive MCF-7 cells and DNA fragmentation. CONCLUSIONS: Our results reveal a substantial tumour-suppressive effect of F7 in the breast cancer model. We propose that the effects of phytochemicals present in this fruit extract are responsible for observed potent anti-cancer activities.

• Klíčová slova
• Angiogenesis, Apoptosis, Cell proliferation, Fruit polyphenols, MCF-7, Mammary carcinogenesis, Rat,
• MeSH
• antigen Ki-67 genetika   metabolismus   MeSH
• apoptóza účinky léků   MeSH
• experimentální nádory mléčných žláz farmakoterapie   MeSH
• flavonoidy analýza   farmakologie   MeSH
• fragmentace DNA účinky léků   MeSH
• fytogenní protinádorové látky analýza   farmakologie   MeSH
• kaspasa 3 genetika   metabolismus   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• methylnitrosomočovina toxicita   MeSH
• MFC-7 buňky MeSH
• modely nemocí na zvířatech MeSH
• ovoce chemie   MeSH
• polyfenoly analýza   farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• protein X asociovaný s bcl-2 genetika   metabolismus   MeSH
• receptor 2 pro vaskulární endoteliální růstový faktor genetika   metabolismus   MeSH
• stilbeny analýza   farmakologie   MeSH
• tyrosin analogy a deriváty   metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 3-nitrotyrosine MeSH Prohlížeč
• antigen Ki-67 MeSH
• Bax protein, rat MeSH Prohlížeč
• Casp3 protein, rat MeSH Prohlížeč
• Flavin7 MeSH Prohlížeč
• flavonoidy MeSH
• fytogenní protinádorové látky MeSH
• kaspasa 3 MeSH
• Kdr protein, rat MeSH Prohlížeč
• methylnitrosomočovina MeSH
• polyfenoly MeSH
• protein X asociovaný s bcl-2 MeSH
• receptor 2 pro vaskulární endoteliální růstový faktor MeSH
• stilbeny MeSH
• tyrosin MeSH