We explored the mechanism of human osteosarcoma MG-63 cell apoptosis induced by asta-xanthin. The MTT assay was used to detect the effect of astaxanthin on cell viability. Morphological changes associated with apoptosis were observed after DAPI staining. Early and late stages of apoptosis were detected by flow cytometry with annexin V-FITC/PI staining. Activation of caspases-8, -9 and -3 was detected by enzyme activity in vitro. Changes in the mitochondrial membrane potential were detected by MitoCapture staining. Western blot was used to detect the cleavage of PARP, which is a caspase-3 substrate, the release of cytochrome c and Smac into the cytosol, the translocation of pro-apoptotic proteins Bax and Bak, and the expression of mitochondrial pathway-related proteins. The translocation of Bax was also detected by immunofluorescence assay. Astaxanthin significantly inhibited the viability of human osteosarcoma MG-63 cells with an IC50 value of 12.36 μg/ml. The DAPI-stained cells showed characteristic apoptotic morphological changes - cell shrinkage, cell membrane blebbing, nuclear condensation, and apoptotic body formation. Cytochrome c and Smac were released from mitochondria to the cytosol. Pro-apoptotic proteins Bax and Bak were rapidly translocated to mitochondria after six hours of astaxanthin action. Caspases-9 and -3 were activated and PARP was cleaved. The expression of anti-apoptotic proteins Bcl-2, Bcl-xL and XIAP was significantly decreased. Astaxanthin induced human osteosarcoma MG-63 cell apoptosis through the mitochondria-mediated endogenous apoptosis pathway.

• MeSH
• apoptóza MeSH
• cytochromy c * metabolismus   MeSH
• lidé MeSH
• osteosarkom * farmakoterapie   metabolismus   MeSH
• PARP inhibitory farmakologie   terapeutické užití   MeSH
• protein X asociovaný s bcl-2 metabolismus   MeSH
• proteiny regulující apoptózu farmakologie   terapeutické užití   MeSH
• protoonkogenní proteiny c-bcl-2 metabolismus   MeSH
• xanthofyly MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: Most aspects of osteosarcoma have been addressed in detail, but there is no comprehensive analysis of deceased patients and causes of death. METHODS: The database of the Cooperative Osteosarcoma Study Group COSS (1980-03/31/2021; 4475 registered high-grade central osteosarcoma patients) was searched deaths from any cause. Affected patients were analyzed for demographic and baseline variables and disease-status at the time of demise. Deaths from causes other than osteosarcoma were analyzed in detail. RESULTS: A total of 1520 deceased patients were identified (median age (range) at osteosarcoma diagnosis 16 (2-78) years; 908 (59.7%) male, 612 (40.3%) female; primary tumor: extremities 1263 (83.1%), trunk 208 (13.7%), craniofacial 47 (3.1%) (site unknown 2); metastases at registration: absent 1.051 (69.1%), present 466 (30.7%) (3 no data). The median time from diagnosis to death was 2.22 (0.08-32.02) years. 1286 (84.6%) patients succumbed to osteosarcoma (370 without achieving complete remission, 488 first, 428 more than one recurrences), 146 (9.6%) to other, 88 (5.8%) to unknown causes. Chemotherapy-related infections (40), secondary malignancies (39), and perioperative complications (19) were among the most frequent potentially treatment-related causes, and high-dose methotrexate (19), doxorubicin (17), and ifosfamide (15) were the drugs most commonly held responsible. Patients with unknown causes of death had an unusually long median follow-up. CONCLUSION: The major cause of death of patients after osteosarcoma is this malignancy, mostly from one of its multiple relapses. However, almost 10% of fatalities are due to other documented causes. Some of these deaths may be preventable with the knowledge gained from comprehensive analyses such as this.

• MeSH
• cisplatina terapeutické užití   MeSH
• doxorubicin terapeutické užití   MeSH
• ifosfamid terapeutické užití   MeSH
• lidé MeSH
• lokální recidiva nádoru MeSH
• methotrexát MeSH
• mladiství MeSH
• nádory kostí * farmakoterapie   MeSH
• osteosarkom * farmakoterapie   MeSH
• příčina smrti MeSH
• protokoly protinádorové kombinované chemoterapie MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Klíčové buněčné procesy včetně proliferace, diferenciace a přežívání jsou často regulovány stejnými molekulami během ontogeneze a tumorigeneze. Je známo, že protein c-Myb se spolupodílí na řízení uvedených procesů. Aberantní aktivace tohoto proteinu narušuje rovnováhu v jejich regulaci a podporuje zvýšenou proliferaci a přežívání buněk, čímž přispívá k maligní transformaci. Náš předchozí výzkum ukázal, že c-Myb se podílí také na regulaci fyziologického vývoje kostí. Nedávno publikované výsledky, podpořené našimi preliminárními daty, potvrdily, že protein c-Myb je exprimován v osteosarkomech, nádorech kostní tkáně, které se vyznačují věkově specifickou incidencí. Cílem tohoto projektu je objasnění funkce proteinu c-Myb v různých fázích fyziologického vývoje kostí a především jeho významu v regulaci progrese a chemorezistence osteosarkomů. s využitím buněčných linií, in vivo modelů a klinické studie. Klinická část bude dále rozšířena o širší spektrum osteogenních patologií s typickým výskytem v období dospívání, což přispěje ke zhodnocení významu proteinu c-Myb v homeostáze kostí.; Ontogenesis and tumorigenesis often share regulatory molecules that modulate key cellular events including cell proliferation, differentiation and survival. The c-Myb protein has been associated with control of these processes. Aberrant activation of c-Myb disturbs the balance in favor of proliferation and survival, thus contributing to malignant transformation. Our earlier research showed that c-Myb is involved in regulation of embryonal bone development. Recent findings, including preliminary data presented by us, demonstrate that c-Myb is expressed in osteosarcomas, bone tumors with specific age-related incidence. The aim of this study is to clarify the function of c-Myb in different stages of physiological bone development, and most importantly, the relevance of c-Myb in control of osteosarcoma progression and chemoresistance using cell lines, in vivo models and a clinical study. The clinical part will be further extended to other osteogenic pathologies with high incidence in childhood to further assess the importance of c-Myb in bone homeostasis in humans.

• MeSH
• chemorezistence MeSH
• geny myb MeSH
• homeostáza MeSH
• karcinogeneze genetika   MeSH
• mladiství MeSH
• osteogeneze genetika   MeSH
• osteosarkom farmakoterapie   genetika   MeSH
• Check Tag
• mladiství MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• genetika, lékařská genetika
• osteologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Osteosarcoma (OS), a severe malignant bone tumour, usually occurs in adolescents and children and has a poor prognosis. Asiatic acid (AA), an active component isolated from Centella asiatica (L.) Urb., exhibits appreciable anti-oxidant and anti-tumour activities. So far, the effects and underlying mechanisms of AA against OS have not been clarified. Here, we explored the anti-tumour effects of AA against human OS and the involved mechanism mediating its actions. To evaluate effects of AA on the cell proliferation of human OS cells, cell viability and colony formation assays were performed. Flow cytometry was used to evaluate apoptosis in OS cells exposed to AA and mitochondrial membrane potential. Western blotting and RT-PCR were applied to determine expression of the relevant proteins and their mRNA levels. Our explorations showed that AA inhibits proliferation of human OS cells in a concentration- and time-dependent manner, and induces apoptosis of OS cells by the intrinsic (mitochondrial) pathway. Importantly, we found that inhibition of the AA-induced phosphorylation of JAK2/STAT3 signalling molecules and the decrease in MCL-1 contributed to the anti-tumour efficacy of AA. Collectively, our results suggest that AA could evoke mitochondrial- induced apoptosis in human OS cells by suppression of the JAK2/STAT3 pathway and MCL-1 expression. These results strongly demonstrate that AA could be a potential anti-tumour agent for OS treatment.

• MeSH
• apoptóza * MeSH
• Janus kinasa 2 MeSH
• lidé MeSH
• mladiství MeSH
• nádorové buněčné linie MeSH
• osteosarkom * farmakoterapie   MeSH
• pentacyklické triterpeny farmakologie   MeSH
• proliferace buněk MeSH
• transkripční faktor STAT3 MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• Publikační typ
• časopisecké články MeSH

NF-κB is activated in a variety of human cancers. However, its role in osteosarcoma (OS) remains unknown. Here, we have elucidated the implication of NF-κB in the oncogenic phenotype of OS tumor cells. We reported that activation of NF-κB was a common event in the human OS. Inhibition of NF-κB using inhibitor Bay 11-7085 repressed proliferation, survival, migration, and invasion but increased apoptosis in 143B and MG63 OS cells, indicating that NF-κB is critically implicated in the oncogenesis of OS. Notably, Bay 11-7085 not only inactivated NF-κB but also reduced the phosphorylation of AKT via its induction of PTEN, suggesting the existence of a novel NF-κB/PTEN/PI3K/AKT axis. In vivo, Bay 11-7085 suppressed tumor growth in the bone by targeting NF-κB and AKT. Interestingly, combined treatment with Bay 11-7085 and the PI3K inhibitor, LY294002, triggered an augmented antitumor effect. Our results demonstrate that NF-κB potentiates the growth and aggressiveness of OS. Pharmacological inhibition of NF-κB represents a promising therapy for the treatment of OS.

• Klíčová slova
• Bay 11-7085 Sigma-Aldrich,
• MeSH
• cílená molekulární terapie MeSH
• dimethylsulfoxid terapeutické užití   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední NOD MeSH
• nádorové buňky kultivované účinky léků   MeSH
• NF-kappa B účinky léků   MeSH
• nitrily farmakologie   terapeutické užití   MeSH
• osteosarkom * farmakoterapie   genetika   patologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• protoonkogenní proteiny c-akt antagonisté a inhibitory   genetika   metabolismus   MeSH
• RNA nádorová izolace a purifikace   účinky léků   MeSH
• sulfony farmakologie   terapeutické užití   MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

The main objective of this study was to analyze changes in the antiproliferative effect of vitamin D3, in the form of calcitriol and calcidiol, via its combined application with all-trans retinoic acid (ATRA) in osteosarcoma cell lines. The response to treatment with calcitriol and calcidiol alone was specific for each cell line. Nevertheless, we observed an enhanced effect of combined treatment with ATRA and calcitriol in the majority of the cell lines. Although the levels of respective nuclear receptors did not correlate with the sensitivity of cells to these drugs, vitamin D receptor (VDR) upregulation induced by ATRA was found in cell lines that were the most sensitive to the combined treatment. In addition, all these cell lines showed high endogenous levels of retinoic acid receptor α (RARα). Our study confirmed that the combination of calcitriol and ATRA can achieve enhanced antiproliferative effects in human osteosarcoma cell lines in vitro. Moreover, we provide the first evidence that ATRA is able to upregulate VDR expression in human osteosarcoma cells. According to our results, the endogenous levels of RARα and VDR could be used as a predictor of possible synergy between ATRA and calcitriol in osteosarcoma cells.

• MeSH
• alfa receptor kyseliny retinové metabolismus   MeSH
• kalcifediol aplikace a dávkování   MeSH
• kalcitriol aplikace a dávkování   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• osteosarkom farmakoterapie   metabolismus   MeSH
• protinádorové látky aplikace a dávkování   MeSH
• protokoly protinádorové kombinované chemoterapie MeSH
• receptory kalcitriolu metabolismus   MeSH
• screeningové testy protinádorových léčiv MeSH
• tretinoin aplikace a dávkování   MeSH
• vitaminy aplikace a dávkování   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Some epidemiological studies suggested caffeine consumption as the cause for bone mineral density loss. Certain genes involved in this process are regulated by vitamin D receptor (VDR). Therefore, we investigated if caffeine can affect inducible expression of VDR-regulated genes, some of them being involved in bone mineralization process. By employing reporter gene assay, polymerase chain reaction, and western blotting, we monitored the VDR activity and expression in cell cultures of intestinal (LS180), osteosarcoma (HOS), and normal human osteoblasts in vitro. While caffeine stimulated calcitriol-inducible VDR-dependent nanoluciferase activity in stable reporter cell line IZ-VDRE (derived from LS180), it rather modulated mRNA levels of target genes, like CYP24A1, BGLAP, SPP1, and TNSF11 in LS180 and HOS cells. However, caffeine significantly decreased calcitriol-inducible CYP24A1, TNSF11, and SPP1 transcripts in osteoblasts. This decrease had non-linear U-shaped profile. Our in vitro data demonstrate biphasic action of caffeine on the expression of certain calcitriol-inducible VDR-regulated genes in normal human osteoblasts.

• MeSH
• kalcitriol farmakologie   MeSH
• kofein farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• osteoblasty účinky léků   metabolismus   MeSH
• osteosarkom farmakoterapie   metabolismus   MeSH
• receptory kalcitriolu účinky léků   metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• signální transdukce účinky léků   MeSH
• vitamin D metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• chondrosarkom epidemiologie   farmakoterapie   chirurgie   radioterapie   MeSH
• chordom epidemiologie   chirurgie   radioterapie   MeSH
• cytostatické látky terapeutické užití   MeSH
• Ewingův sarkom epidemiologie   farmakoterapie   chirurgie   sekundární   MeSH
• kombinovaná farmakoterapie MeSH
• lidé MeSH
• nádory kostí * epidemiologie   farmakoterapie   chirurgie   MeSH
• neoadjuvantní terapie MeSH
• osteosarkom epidemiologie   farmakoterapie   chirurgie   sekundární   MeSH
• sarkom * diagnóza   farmakoterapie   chirurgie   klasifikace   MeSH
• staging nádorů metody   MeSH
• věkové skupiny MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

BACKGROUND: High-grade osteosarcoma is a primary malignant bone tumour mainly affecting children and young adults. The European and American Osteosarcoma Study (EURAMOS)-1 is a collaboration of four study groups aiming to improve outcomes of this rare disease by facilitating randomised controlled trials. METHODS: Patients eligible for EURAMOS-1 were aged ≤40 years with M0 or M1 skeletal high-grade osteosarcoma in which case complete surgical resection at all sites was deemed to be possible. A three-drug combination with methotrexate, doxorubicin and cisplatin was defined as standard chemotherapy, and between April 2005 and June 2011, 2260 patients were registered. We report survival outcomes and prognostic factors in the full cohort of registered patients. RESULTS: For all registered patients at a median follow-up of 54 months (interquartile range: 38-73) from biopsy, 3-year and 5-year event-free survival were 59% (95% confidence interval [CI]: 57-61%) and 54% (95% CI: 52-56%), respectively. Multivariate analyses showed that the most adverse factors at diagnosis were pulmonary metastases (hazard ratio [HR] = 2.34, 95% CI: 1.95-2.81), non-pulmonary metastases (HR = 1.94, 95% CI: 1.38-2.73) or an axial skeleton tumour site (HR = 1.53, 95% CI: 1.10-2.13). The histological subtypes telangiectatic (HR = 0.52, 95% CI: 0.33-0.80) and unspecified conventional (HR = 0.67, 95% CI: 0.52-0.88) were associated with a favourable prognosis compared with chondroblastic subtype. The 3-year and 5-year overall survival from biopsy were 79% (95% CI: 77-81%) and 71% (95% CI: 68-73%), respectively. For patients with localised disease at presentation and in complete remission after surgery, having a poor histological response was associated with worse outcome after surgery (HR = 2.13, 95% CI: 1.76-2.58). In radically operated patients, there was no good evidence that axial tumour site was associated with worse outcome. CONCLUSIONS: In conclusion, data from >2000 patients registered to EURAMOS-1 demonstrated survival rates in concordance with institution- or group-level osteosarcoma trials. Further efforts are required to drive improvements for patients who can be identified to be at higher risk of adverse outcome. This trial reaffirms known prognostic factors, and owing to the large numbers of patients registered, it sheds light on some additional factors to consider.

• MeSH
• cisplatina aplikace a dávkování   MeSH
• dítě MeSH
• dospělí MeSH
• doxorubicin aplikace a dávkování   MeSH
• kohortové studie MeSH
• lidé MeSH
• metastázy nádorů MeSH
• methotrexát aplikace a dávkování   MeSH
• míra přežití MeSH
• mladiství MeSH
• nádory kostí farmakoterapie   mortalita   sekundární   MeSH
• následné studie MeSH
• osteosarkom farmakoterapie   mortalita   patologie   MeSH
• prognóza MeSH
• protokoly protinádorové kombinované chemoterapie terapeutické užití   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze III MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie MeSH

In the present investigation Hibiscus rosa-sinensis (petals), Acorus calamus (rhizome), Moringa oleifera (leaves) and Cucurbita maxima (petals) were screened for their efficacy against osteosarcoma with elucidating a mechanism of their anticancer potentiality. The methanolic plant extracts revealed the presence of all major phytochemicals with quantitative analysis of flavonoids (98.15 ± 2.02 to 12.34 ± 0.57 mg of RUE/g) and total phenolics (26.40 ± 0.11 to 8.54 ± 0.10 mg of GAE/g). The antioxidant activity was assessed by standard DPPH, H2O2 scavenging, NO scavenging assays. The hemolysis, hemagglutination, and erythrocyte aggregation assays unveiled their compatibility with blood components. As most of the opportunistic microbes infect subsequently immunocompromised patients, the antimicrobial activity of the plant extracts showed a zone of inhibition (in mm) against nosocomial strains of Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Vibrio cholerae having MIC between 12.5–50 μg/ml. Through MTT assay the IC50 value was calculated against MG 63 osteosarcoma with detailed studies on DNA fragmentation and chromatin condensation, revealing apoptosis being their primary mode of anticancer effect. Further the migration and colony forming assays supported the anticancer potentials of the methanolic plant extracts. The cell cycle analysis revealed that A. calamus and M. oleifera extracts were capable of arresting the growth of MG 63 osteosarcoma cells.

• MeSH
• antioxidancia MeSH
• Cucurbita chemie   MeSH
• fytogenní protinádorové látky * farmakologie   chemie   MeSH
• fytoterapie MeSH
• Hibiscus chemie   MeSH
• léčivé rostliny MeSH
• mikrobiální testy citlivosti MeSH
• Moringa chemie   MeSH
• nádorové buňky kultivované účinky léků   MeSH
• osteosarkom farmakoterapie   MeSH
• puškvorec chemie   MeSH
• rostlinné extrakty MeSH
• screeningové testy protinádorových léčiv MeSH
• techniky in vitro MeSH