The mechanisms of nadroparin-mediated inhibition of proliferation of two human lung cancer cell lines.

OBJECTIVES: Clinical data suggest that heparin treatment improves survival of lung cancer patients, but the mechanisms involved are not fully understood. We investigated whether low molecular weight heparin nadroparin, directly affects lung cancer cell population growth in conventionally cultured cell lines. MATERIALS AND METHODS: A549 and CALU1 cells' viability was assessed by MTT and trypan blue exclusion assays. Cell proliferation was assessed using 5-bromo-2-deoxyuridine incorporation. Apoptosis and cell-cycle distribution were analysed by flow cytometry; cyclin B1, Cdk1, p-Cdk1 Cdc25C, p-Cdc25C and p21 expressions were analysed by western blotting. mRNA levels were analysed by real time RT-PCR. RESULTS: Nadroparin inhibited cell proliferation by 30% in both cell lines; it affected the cell cycle in A549, but not in CALU-1 cells, inducing arrest in the G(2) /M phase. Nadroparin in A549 culture inhibited cyclin B1, Cdk1, Cdc25C and p-Cdc25C, while levels of p-Cdk1 were elevated; p21 expression was not altered. Dalteparin caused a similar reduction in A549 cell population growth; however, it did not alter cyclin B1 expression as expected, based on previous reports. Fondaparinux caused minimal inhibition of A549 cell population growth and no effect on either cell cycle or cyclin B1 expression. CONCLUSIONS: Nadroparin inhibited proliferation of A549 cells by inducing G(2) /M phase cell-cycle arrest that was dependent on the Cdc25C pathway, whereas CALU-1 cell proliferation was halted by as yet not elucidated modes.

Role of NF-kappaB and PPAR-gamma in lung inflammation induced by monocyte-derived microparticles.

Microparticles (MP) are phospholipid vesicles shed by cells upon activation or apoptosis. Monocyte-derived MP upregulate the synthesis of proinflammatory mediators by lung epithelial cells; the molecular bases of such activity are unknown. Peroxisome proliferator-activated receptors (PPAR) have been demonstrated to be involved in the modulation of nuclear factor (NF)-κB transcriptional activity and inflammation. We investigated whether the upregulation of the synthesis of proinflammatory cytokines by human lung epithelial cells induced by monocyte/macrophage-derived MP involves NF-κB activation and is modulated by PPAR-γ. MP were generated by stimulation of human monocytes/macrophages with the calcium ionophore, A23187. MP were incubated with human lung epithelial cells. NF-κB translocation was assessed by electrophoretic mobility shift assay. Interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 synthesis was assessed by ELISA and RT-PCR. Stimulation of A549 alveolar cells with monocyte/macrophage-derived MP caused an increase in NF-κB activation and IL-8 and MCP-1 synthesis that was inhibited by pre-incubation with the PPAR-γ agonists, rosiglitazone and 15-deoxy-Δ12,14-prostaglandin-J2. Parallel experiments with normal human bronchial epithelial cells largely confirmed the results. The effects of PPAR-γ agonists were reversed by the specific antagonist, GW9662. Upregulation of the synthesis of proinflammatory mediators by human lung epithelial cells induced by monocyte/macrophage-derived MP is mediated by NF-κB activation through a PPAR-γ dependent pathway.

A histochemical approach to the evaluation of the in vivo cytotoxicity of the nitrobutadienes (1E,3E)-1,4-bis(1-naphthyl)-2,3-dinitro-1,3-butadiene and methyl (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate in mice liver and kidney.

Two new molecules (1E,3E)-1,4-bis(1-naphthyl)-2,3-dinitro-1,3-butadiene (1-Naph-DNB) and (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate (1-Naph-NMCB) in previous studies showed interesting antiproliferative activity in vitro. Furthermore, toxicological tests and histological analysis provided promising results, in particular for 1-Naph-NMCB that displayed lower toxic activity both in terms of lethal effect and tissue damage of the main organs. Finally, studies of the antitumour activity in vivo confirmed the efficacy of both molecules, though with some differences in tumour selectivity and levels of activity. In this investigation the activities of some specific enzymes, acid phosphatase (AcPase), alkaline phosphatase (AlkPase), catalase (Cat), succinic dehydrogenase (SDH), glucose-6-phosphatase (G6Pase) and K+ p-nitrophenyl phosphatase (K+ pNPPase) were studied in the liver and kidney as histopathological biomarkers, to assess the effects of the two compounds in organs generally involved in the metabolism and excretion of different drugs. As oxidative stress may also develop as a consequence of the toxic effect of chemicals, reactive oxygen species (ROS) production was evaluated by a histochemical method. The results indicated that some enzyme activities and ROS expression changed in a dose-related manner. Nevertheless, neither in the liver nor in the kidney were dramatic toxic effects evident. By contrast, the variations of some enzyme activities (AlkPase, AcPase, Cat, K+ pNPPase) were interpreted as possible defensive mechanisms for tolerating high dosage of the compounds.

Effect of seed mass on germination and growth in three dominant species in southern Brazilian coastal dunes

The effect of seed mass on germination and growth was tested in fresh-seeds of Blutaparon portulacoides, Panicum racemosum, and Spartina ciliata, selected at random in southern Brazilian populations. The seed mass varied within a population of the three species. Both B. portulacoides and P. racemosum showed normal frequency distribution of seed mass, while S. ciliata did not. Significant differences were observed in seed germination between large and small seeds of all species. In all species the capacity of seedling elongation was greater in seedlings of large seeds than those of small ones. Relative growth rate of seedlings of P. racemosum and S. ciliata decreased with time in all seed mass size-classes. On the other hand, the relative growth rate of B. portulacoides seedlings increased during the first 40 days. Seed mass is an important biological factor, affecting seed germination, seedling elongation, and growth of these species, and favoring large seeds, specially in areas of active sand accretion like coastal dunes

Effect of seed mass on germination and growth in three dominant species in southern Brazilian coastal dunes

The effect of seed mass on germination and growth was tested in fresh-seeds of Blutaparon portulacoides, Panicum racemosum, and Spartina ciliata, selected at random in southern Brazilian populations. The seed mass varied within a population of the three species. Both B. portulacoides and P. racemosum showed normal frequency distribution of seed mass, while S. ciliata did not. Significant differences were observed in seed germination between large and small seeds of all species. In all species the capacity of seedling elongation was greater in seedlings of large seeds than those of small ones. Relative growth rate of seedlings of P. racemosum and S. ciliata decreased with time in all seed mass size-classes. On the other hand, the relative growth rate of B. portulacoides seedlings increased during the first 40 days. Seed mass is an important biological factor, affecting seed germination, seedling elongation, and growth of these species, and favoring large seeds, specially in areas of active sand accretion like coastal dunes.

O efeito da massa das sementes sobre a germinação e o crescimento foi testado com sementes de Blutaparon portulacoides, Panicum racemosum e Spartina ciliata coletadas aleatoriamente em populações no sul do Brasil. A massa das sementes variou dentro das populações das três espécies. Ambas, B. portulacoides e P. racemosum, mostraram distribuição normal na freqüência no peso das sementes, enquanto S. ciliata não apresentou distribuição normal. Diferenças significativas foram observadas na germinação das sementes entre sementes grandes e pequenas nas três espécies. A capacidade de alongamento das plântulas das três espécies é maior nas plântulas originadas de sementes grandes do que nas provenientes de sementes pequenas. As taxas de crescimento relativo das plântulas de P. racemosum e S. ciliata diminuíram com o tempo em todas as classes de massa de sementes. Por outro lado, a taxa de crescimento relativo das plântulas de B. portulacoides aumentou ao longo dos primeiros 40 dias. A massa das sementes é um importante fator biológico, porque afeta a germinação, o alongamento das plântulas e o crescimento nas espécies, favorecendo assim as sementes maiores, especialmente em áreas de ativa deposição de areias, como as dunas costeiras.

Effect of seed mass on germination and growth in three dominant species in southern Brazilian coastal dunes.

The effect of seed mass on germination and growth was tested in fresh-seeds of Blutaparon portulacoides, Panicum racemosum, and Spartina ciliata, selected at random in southern Brazilian populations. The seed mass varied within a population of the three species. Both B. portulacoides and P. racemosum showed normal frequency distribution of seed mass, while S. ciliata did not. Significant differences were observed in seed germination between large and small seeds of all species. In all species the capacity of seedling elongation was greater in seedlings of large seeds than those of small ones. Relative growth rate of seedlings of P. racemosum and S. ciliata decreased with time in all seed mass size-classes. On the other hand, the relative growth rate of B. portulacoides seedlings increased during the first 40 days. Seed mass is an important biological factor, affecting seed germination, seedling elongation, and growth of these species, and favoring large seeds, specially in areas of active sand accretion like coastal dunes.