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Platinum‐based chemotherapeutic agents are widely used in the treatment of cancer. However, their effectiveness is limited by severe adverse reactions, drug resistance, and poor water solubility. This study focuses on the synthesis and characterization of new water‐soluble cationic monofunctional platinum(II) complexes starting from the [PtCl(η1‐C2H4OEt)(phen)] (1, phen=1,10‐phenanthroline) precursor, specifically [Pt(NH3)(η1‐C2H4OEt)(phen)]Cl (2), [Pt(1‐hexyl‐1H‐imidazole)(η1‐C2H4OEt)(phen)]Cl (3), and [Pt(1‐hexyl‐1H‐benzo[d]imidazole)(η1‐C2H4OEt)(phen)]Cl (4), which deviate from traditional requirements for antitumor activity. These complexes were evaluated for their cytotoxic effects in comparison to cisplatin, using immortalized cervical adenocarcinoma cells (HeLa), human renal carcinoma cells (Caki‐1), and normal human renal cells (HK‐2). While complex 2 showed minimal effects on the cell lines, complexes 3 and 4 demonstrated higher cytotoxicity than cisplatin. Notably, complex 4 displayed the highest cytotoxicity in both cancer and normal cell lines. However, complex 3 exhibited the highest selectivity for renal tumor cells (Caki‐1) among the tested complexes, compared to healthy cells (HK‐2). This resulted in a significantly higher selectivity than that of cisplatin and complex 4. Therefore, complex 3 shows potential as a leading candidate for the development of a new generation of platinum‐based anticancer drugs, utilizing biocompatible imidazole ligands while demonstrating promising anticancer properties. [ABSTRACT FROM AUTHOR]

There is very little about the impact that sports training has on female football referees. Therefore, we determined the effects of a 40‐week physical preparation, including a full football season, on plasma testosterone and cortisol concentrations and physical performance in female football referees. Plasma cortisol and testosterone concentrations were assayed together with fitness tests at the beginning of the training period (T0, in September), after 8 weeks from T0 (T1), at the mid of the season (T2, 24 weeks after T0), and at the end of the season (T3, in June, 40 weeks after T0). Plasma cortisol increased during the first period and up to T2 (from 15.4 ± 4.7 to 28.5 ± 3.9 µg/dl; p < 0.001), and then decreased at the end of the season (T3: 16.0 ± 2.4 µg/dl). Plasma testosterone concentration in T0 was 14.2±0.37 µg/dl and increased in T1 (57.1 ± 3.7 µg/dl) and T2 (47 ± 3.7 µg/dl) and then decreased in T3 (33.5 ± 2.8 µg/dl). Resting testosterone levels in women were very low (14,2 ± 0.37 µg/dl) (Figure 3c). Testosterone increased in T1 (57.1 ± 3.7 µg/dl) and T2 (47 ± 3.7 µg/dl) whilst, at the end of the season, its concentration decreased (33.5 ± 2.8 µg/dl) (Figure 3c). Significant improvements were observed in all physical performances during the observed period (ANOVA, p < 0.05). Finally, testosterone and cortisol concentrations significantly (p < 0.0001 for both) correlated with maximal oxygen consumption. In T1, testosterone concentration was also significantly correlated with running speed test (p < 0.001). In conclusion, training induces endocrine changes in order to maintain body homeostasis in women referees. It is important that coaches and sports scientists regularly observe changes in endocrine function induced by training and matches in female referees, because they can help maximize referees' performance and limit cases of overtraining. [ABSTRACT FROM AUTHOR]

The global prevalence of childhood obesity is high. Obesity main causes are linked to sedentary lifestyles. Increasing physical activity (PA) and reducing sedentary activities are recommended to prevent and treat obesity. This study aimed to evaluate the effectiveness of a 6‐month school PA intervention on obesity prevention and healthy behaviors in school‐aged children. Participating students (10–11 years of age) were randomly divided into an intervention group and a control group. Children in the intervention group (n = 80) participated in a multicomponent PA that included improvement in extracurricular physical activities (with an additional 40 min per day for 5/6 days per week). Children (n = 80) in the control group participated in usual practice. Participants had mean body mass index of 19.7 ± 2.9 kg/m2, and 33.7% of them were overweight or with obesity at T0. The change in body mass index in intervention group (−2.4 ± 0.6 kg/m2) was significantly different from that in control group (3.01 ± 1.8 kg/m2). The effects on waist circumference, waist‐to‐height ratio, and physic fitness were also significant in intervention group compared with control group (all p < 0.05). Furthermore, there is a significant decrease in overweight or children with obesity in the experimental group (to 17.5%, p < 0.05). These findings suggest that a school‐based intervention program represents an effective strategy for decreasing the number of overweight and children with obesity. [ABSTRACT FROM AUTHOR]

Salvia clandestina L. is a wild perennial species present in the Salento area of Italy. Here, we examined the in vitro effects of an aqueous extract of S. clandestina L. on the MG‐63 osteosarcoma cell line. The extract reduced osteosarcoma cell viability mainly by way of apoptosis, as we observed (1) upregulation of gene and protein expression of p53, cyclin‐dependent kinase inhibitors p21WAF1 and p27Kip1, and proapoptotic BAX; (2) activation of caspases; and (3) induction of a sub‐G1 peak in the cell cycle. The mitogen‐activated protein kinases (MAPKs) JNK1/2 and p38 are activated and involved in the intracellular effects of the S. clandestina extract, as preincubation with the JNK1/2 inhibitor SP600125 or the p38 inhibitor SB203580 significantly decreased S. clandestina extract–induced cytotoxicity and inhibited increase in p53, p21WAF1, p27Kip1, and BAX. SP600125 also inhibited mRNA levels for all the aforementioned proteins, while SB203580 only affected p53 mRNA. Furthermore, S. clandestina extract treatment counteracted epithelial‐to‐mesenchymal transition, inhibited cell migration, and decreased the expression and activity of matrix metalloproteinase MMP2. In addition, S. clandestina extract enhanced the cytotoxic activity of cisplatin on MG‐63 cells through downregulation of the Akt/PKB protein kinase. We conclude that S. clandestina extract may be a novel agent for osteosarcoma treatment. [ABSTRACT FROM AUTHOR]

As no study has explored the impact of physical stress on hypothalamic‐pituitary‐gonadal axis hormones over a long period, the purpose of this study was to determine the effects of the football season period on plasma cortisol and testosterone concentrations and referee's physical performances. Physical tests and plasma cortisol and testosterone concentrations were assayed before the beginning of the training period, just after the training period, at the middle of the season, and at the end of the season, in 29 male football referees and 30 healthy control subjects. Results showed significant differences in hormone concentrations at the four‐time points evaluated. Plasma cortisol increased during the first training period from 15.8 ± 3.8 to 21.7 ± 5.1 µg/dl (p < 0.001), then decreased during the season and at the end of it was 18.7 ± 2.4 µg/dl. Before the beginning of the training period, plasma testosterone concentration was 386.1 ± 58.8 ng/dl; after the training period, it increased to 572.2 ± 88.1 ng/dl (p < 0.001) and then returned to baseline levels at the end of the season. Between the start of the training period and the end of the season, significant differences were observed in physical performances of referees. Plasma cortisol and testosterone levels significantly (p < 0.0001 for both) correlated with Yo‐Yo intermittent recovery test level 1 (YYIRT1) and maximal oxygen consumption (VO2max) at the end of the training period. In the middle season, plasma testosterone concentration only significantly (p < 0.0001) correlated with YYIRT1 and VO2max. These data underline the importance of set up training protocols that present the prospective to favor positive physiological adaptations. [ABSTRACT FROM AUTHOR]

Following peripheral nerve injury, remnant Schwann cells adopt a migratory phenotype and remodel the extracellular matrix allowing axonal regrowth. Although much evidence has demonstrated that TGF‐β1 promotes glioma cell motility and induces the expression of extracellular matrix proteins, the effects of TGF‐β1 on Schwann cell migration has not yet been studied. We therefore investigated the cellular effects and the signal transduction pathways evoked by TGF‐β1 in rattus norvegicus neuronal Schwann RSC96 cell. TGF‐β1 significantly increased migration and invasion of Schwann cells assessed by the wound‐healing assay and by cell invasion assay. TGF‐β1‐enhanced migration/invasion was blocked by inhibition of MMP‐2 and MMP‐9. Consistently, by real‐time and western blot analyses, we demonstrated that TGF‐β1 increased MMP‐2 and MMP‐9 mRNA and protein levels. TGF‐β1 also increased MMPs activities in cell growth medium, as shown by gelatin zymography. The selective TGF‐β Type I receptor inhibitor SB431542 completely abrogated any effects by TGF‐β1. Indeed, TGF‐β1 Type I receptor activation provoked the cytosol‐to‐nucleus translocation of SMAD2 and SMAD3. SMAD2 knockdown by siRNA blocked MMP‐2 induction and cell migration/invasion due to TGF‐β1. TGF‐β1 also provoked phosphorylation of MAPKs extracellular regulated kinase 1/2 and JNK1/2. Both MAPKs were upstream to p65/NF‐kB inasmuch as both MAPKs' inhibitors PD98059 and SP600125 or their down‐regulation by siRNA significantly blocked the TGF‐β1‐induced nuclear translocation of p65/NF‐kB. In addition, p65/NF‐κB siRNA knockdown inhibited the effects of TGF‐β1 on both MMP‐9 and cell migration/invasion. We conclude that TGF‐β1 controls RSC96 Schwann cell migration and invasion through MMP‐2 and MMP‐9 activities. MMP‐2 is controlled by SMAD2 whilst MMP‐9 is controlled via an ERK1/2‐JNK1/2‐NF‐κB dependent pathway. [ABSTRACT FROM AUTHOR]

To test the hypothesis that football training would be accompanied by physiological adaptations and hormonal changes, we analyzed the effects of a whole football season on physical fitness and hormonal concentrations in youth football players. Male football players (n = 29, age 16.51 ± 0.7 years) in a regional professional league and male healthy control subjects (n = 30, age 17.1 ± 1 years) participated to the study. Blood cortisol, testosterone, and growth hormone (hGH) concentrations were assayed before the beginning of the training period (T0), just after the training period (T1), at the middle of the season (T2), and at the end of the season (T3). In each period physical tests and anthropometric measurements were also performed. Results showed significant differences in basal values of cortisol, testosterone, and growth hormone (hGH) in the four time points evaluated (P < 0.01). In addition, the concentrations of hGH were higher in the soccer players group than in control subjects (P < 0.001). Between the start of the training period and the end of the football season significant differences were observed in the anthropometric characteristics and in the physical form of the football players. Furthermore, the hormonal status was significantly correlated with the indicators of the lower limb power (squat‐jump [SqJ], and counter‐movement‐jump [CMJ]) and those of aerobic performance (Yo–Yo intermittent recovery test level 1 (YYIRT1) and maximal oxygen consumption (VO2max)).These data underscore the importance of establishing training protocols that present the potential to promote positive adaptations without, at the same time, provoking overtraining of young players. [ABSTRACT FROM AUTHOR]

Malignant pleural mesothelioma (MPM) is an aggressive malignant tumor in which cisplatin therapy is commonly used, although its effectiveness is limited. It follows that research efforts dedicated to identify promising combinations that can synergistically kill cancer cells are needed. Because we recently demonstrated that ADP inhibits the proliferation of ZL55 cells, an MPM‐derived cell line obtained from bioptic samples of asbestos‐exposed patients. Our objective in this study was to investigate the hypothesis that ADP also potentiates the cytotoxic activity of cisplatin. Results show that in ZL55 cells ADP enhanced (a) the cytotoxicity of cisplatin by 12‐fold, (b) the restraint of cell clonogenic potential cisplatin‐mediated, and (c) the number of apoptotic cells. Cisplatin, but not ADP, caused caspases activation; nevertheless, poly(ADP‐ribose) polymerase‐1 was not only cleaved in cisplatin‐treated cells but also in cells treated with ADP alone. Furthermore, ADP, but not cisplatin, decreased mTOR and 6SK phosphorylations. Both ADP and cisplatin increased p53 protein, but ADP was also able to enhance p53 messenger RNA. P53 silencing resulted in a very large decrement of cell death induced by ADP or by cisplatin and reverted ADP effects on mTOR/S6K phosphorylation, suggesting that activated p53 may act as a negative regulator of mTOR. Consistently, the inhibition of mTOR by rapamycin also sensitized cells to cisplatin, and the effects of cisplatin plus rapamycin were identical to those obtained with cisplatin plus ADP. These findings suggest that the combination of ADP and cisplatin may be a promising strategy for the clinical treatment of cisplatin‐resistant MPM. The manuscript investigates the effects of ADP on cisplatin treatment in ZL55 epitheliod malignant mesothelioma cells. We show that the simultaneous addition of ADP enhanced the antiproliferative potency of cisplatin with an additive effect of the anticancer drugs and ADP on ZL55 cell apoptosis. In addition, for the first time, we show that extracellular ADP operates a tumor‐suppressive regulatory program by p53, in which cell‐cycle arrest is made mainly at the transcriptional level, whereas cell growth inhibition is enforced by repression of mTOR pathway. [ABSTRACT FROM AUTHOR]

Although an association between cancer progression and matrix metalloproteinase (MMP) 2 and MPP9 expression has been known, the expression, nuclear localization, and physiologically controlled activation of these two MMPs have not been investigated in malignant mesothelioma cells. We examined the expression and intracellular localization of MMP2/9 in ZL55 malignant mesothelioma cells, as well as their regulation by ADP. Using real‐time PCR, we showed that activation of the P2Y1 receptor by ADP increased the expression of MMP2/9 mRNAs; MMP2/9 collected from conditioned media also showed an increase in activity; and ADP induced the nuclear localization of MMP2/9. The effects of ADP on transcription of the MMPs were due to activation of c‐Src, Akt, and NF‐κB, while ERK1/2 phosphorylation was needed for the increase in enzymatic activity and the regulation of nuclear import. We also showed that the nuclear localization of MMP2/9 induced by ADP causes the cleavage and inactivation of poly‐ADP‐ribose polymerase‐1. These findings may help to elucidate the mechanisms regulating MMP2/9 activation in ZL55 human epithelioid mesothelioma cells, and perhaps other cells. Therapeutic approaches that promote ADP accumulation in a tumor environment may constitute an effective means to induce anticancer activity. This paper examines the expression, intracellular localization, and function of matrix metalloproteinase (MMP) 2 and MMP9 in ZL55 human epithelioid mesothelioma cells, as well as their regulation by ADP. We found that ADP activates an intracellular transduction system based on NF‐κB, c‐Src, Akt, and ERK1/2 that causes nuclear localization of MMP2/9, resulting in degradation of the DNA repair enzyme poly‐ADP‐ribose polymerase‐1. [ABSTRACT FROM AUTHOR]

Extracellular nucleotides can regulate cell proliferation in both normal and tumorigenic tissues. Here, we studied how extracellular nucleotides regulate the proliferation of ZL55 cells, a mesothelioma-derived cell line obtained from bioptic samples of asbestos-exposed patients. ADP and 2-MeS-ADP inhibited ZL55 cell proliferation, whereas ATP, UTP, and UDP were inactive. The nucleotide potency profile and the blockade of the ADP-mediated inhibitory effect by the phospholipase C inhibitor U-73122 suggest that P2Y1 receptor controls ZL55 cell proliferation. The activation of P2Y1 receptor by ADP leads to activation of intracellular transduction pathways involving [Ca2+]i, PKC-δ/PKC-α, and MAPKs, ERK1/2 and JNK1/2. Cell treatment with ADP or 2-MeS-ADP also provokes the activation of p53, causing an accumulation of the G1 cyclin-dependent kinase inhibitors p21WAF1 and p27Kip. Inhibition of ZL55 cell proliferation by ADP was completely reversed by inhibiting MEK1/2, or JNK1/2, or PKC-δ, and PKC-α. Through the inhibition of ADP-activated transductional kinases it was found that PKC-δ was responsible for JNK1/2 activation. JNK1/2 has a role in transcriptional up-regulation of p53, p21WAF1/CIP1, and p27kip1. Conversely, the ADP-activated PKC-α provoked ERK1/2 phosphorylation. ERK1/2 increased p53 stabilization, required to G1 arrest of ZL55 cells. Concluding, the importance of the study is twofold: first, results shed light on the mechanism of cell cycle inhibition by ADP; second, results suggest that extracellular ADP may inhibit mesothelioma progression. [ABSTRACT FROM AUTHOR]

Background and Purpose: The aim of this study was to determine whether [platinum (Pt)(O,O'-acetylacetonate (acac))(γ-acac)(dimethylsulphide (DMS))] is differentially cytotoxic in normal and cancer cells, and to measure comparative levels of cytotoxicity compared with cisplatin in the same cells.Experimental Approach: We performed experiments on cancerous and normal epithelial breast cells in primary culture obtained from the same patients. The apoptotic effects [Pt(O,O'-acac)(γ-acac)(DMS)] and cisplatin in cancerous and normal breast cells were compared.Key Results: Cancer cells were more sensitive to [Pt(O,O'-acac)(γ-acac)(DMS)] (IC50 = 5.22 ± 1.2 μmol·L(-1)) than normal cells (IC50 = 116.9 ± 8.8 μmol·L(-1)). However, the difference was less strong when cisplatin was used (IC50 = 96.0 ± 6.9 and 61.9 ± 6.1 μmol·L(-1) for cancer and normal cells respectively). Both compounds caused reactive oxygen species (ROS) production with different mechanisms: [Pt(O,O'-acac)(γ-acac)(DMS)] quickly activated NAD(P)H oxidase while cisplatin caused a slower formation of mitochondrial ROS. Cisplatin and [Pt(O,O'-acac)(γ-acac)(DMS)] caused activation of caspases, proteolysis of PARP and modulation of Bcl-2, Bax and Bid. [Pt(O,O'-acac)(γ-acac)(DMS)] also caused leakage of cytochrome c from the mitochondria. Overall, these processes proceeded more quickly in cells treated with [Pt(O,O'-acac)(γ-acac)(DMS)] compared with cisplatin. [Pt(O,O'-acac)(γ-acac)(DMS)] effects were faster and quantitatively greater in cancer than in normal cells. [Pt(O,O'-acac)(γ-acac)(DMS)] caused a fast decrease of mitochondrial membrane potential, especially in cancer cells.Conclusions and Implications: [Pt(O,O'-acac)(γ-acac)(DMS)] was specific to breast cancer cells in primary culture, and this observation makes this compound potentially more interesting than cisplatin. [ABSTRACT FROM AUTHOR]

Background and Purpose The aim of this study was to determine whether [platinum (Pt)( O , O ′-acetylacetonate (acac))(γ-acac)(dimethylsulphide (DMS))] is differentially cytotoxic in normal and cancer cells, and to measure comparative levels of cytotoxicity compared with cisplatin in the same cells. Experimental Approach We performed experiments on cancerous and normal epithelial breast cells in primary culture obtained from the same patients. The apoptotic effects [ Pt( O , O ′-acac)( γ-acac)( DMS)] and cisplatin in cancerous and normal breast cells were compared. Key Results Cancer cells were more sensitive to [ Pt( O , O ′-acac)( γ-acac)( DMS)] ( IC50 = 5.22 ± 1.2 μmol·L−1) than normal cells ( IC50 = 116.9 ± 8.8 μmol·L−1). However, the difference was less strong when cisplatin was used ( IC50 = 96.0 ± 6.9 and 61.9 ± 6.1 μmol·L−1 for cancer and normal cells respectively). Both compounds caused reactive oxygen species ( ROS) production with different mechanisms: [ Pt( O , O ′-acac)( γ-acac)( DMS)] quickly activated NAD( P) H oxidase while cisplatin caused a slower formation of mitochondrial ROS. Cisplatin and [ Pt( O , O ′-acac)( γ-acac)( DMS)] caused activation of caspases, proteolysis of PARP and modulation of Bcl-2, Bax and Bid. [ Pt( O , O ′-acac)( γ-acac)( DMS)] also caused leakage of cytochrome c from the mitochondria. Overall, these processes proceeded more quickly in cells treated with [ Pt( O , O ′-acac)( γ-acac)( DMS)] compared with cisplatin. [ Pt( O , O ′-acac)( γ-acac)( DMS)] effects were faster and quantitatively greater in cancer than in normal cells. [ Pt( O , O ′-acac)( γ-acac)( DMS)] caused a fast decrease of mitochondrial membrane potential, especially in cancer cells. Conclusions and Implications [ Pt( O , O ′-acac)( γ-acac)( DMS)] was specific to breast cancer cells in primary culture, and this observation makes this compound potentially more interesting than cisplatin. [ABSTRACT FROM AUTHOR]

The communication between the tumor cells and the surrounding cells helps drive the process of tumor progression. Since the microenvironment of breast cancer includes CCL20 chemokine, the purpose of this study was to determine whether CCL20 modulates the physiology of healthy breast epithelial cells in areas adjacent to the tumor. Therefore, primary cultures of mammary cells taken from normal peritumoral areas were used. We assessed that breast cells expressed CCR6 CCL20 receptor. Using molecular (siRNA) and pharmacological (inhibitors) techniques, we found multiple signaling kinases to be activated by CCR6 and involved in CCL20-induced breast cell proliferation and migration. The binding of 10 ng/ml CCL20 to CCR6 induced cell migration whilst higher concentrations (from 15 to 25 ng/ml) led to cell proliferation. CCL20 controlled cell migration and MMP-9 expression by PKC-alpha that activated Src, which caused the activation of downstream Akt, JNK, and NF-kB pathways. Furthermore, higher CCL20 concentrations increased cycE and decreased p27Kip expression ending in enhanced cell proliferation. Cell proliferation occurred through PKC-epsilon activation that transactivated EGFR and ERK1/2/MAPK pathway. Although activated by different CCL20 concentrations, these pathways function in parallel and crosstalk to some extent, inasmuch as Akt activation was responsible for ERK1/2 nuclear translocation and enhanced the transcription of of c-fos and c-myc, involved in cell proliferation. In summary, tumor cells exchange signals with the surrounding healthy cells modifying the extracellular matrix through enzyme secretion; thus, CCL20 might be a factor involved in the ontogeny of breast carcinoma. J. Cell. Physiol. 228: 1873-1883, 2013. © 2013 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Liver is an important target for thyroid hormone actions. T3 exerts its effects by two mechanisms: (i) Genomic actions consisting of T3 link to nuclear receptors that bind responsive elements in the promoter of target genes, (ii) non-genomic actions including integrin αvb3 receptor-mediated MAPK/ERK and PI3K/Akt/mTOR-C1 activation. SREBP-1a, SREBP-1c, and SREBP-2 are transcription factors involved in the regulation of lipogenic genes. We show in Hep G2 cells that T3 determined a dose- and time-dependent increase in the level of the precursor form of SREBP-1 without affecting SREBP-1 mRNA abundance. T3 also induced phosphorylation of ERK1/2, Akt and of mTOR-C1 target S6K-P70, and the cytosol-to-membrane translocation of PKC-α. Modulation of SREBP-1 protein level by T3 was dependent on MAPK/ERK, PI3K/Akt/mTOR-C1 pathway activation since the MEK inhibitor PD98059 or the PI3K inhibitor LY294002 abolished the stimulatory effect of T3. Conversely, the effect of T3 on SREBP-1 level was enhanced by using rapamycin, mTOR-C1 inhibitor. These data suggest a negative control of mTOR-C1 target S6K-P70 on PI3K/Akt pathway. The effect of T3 on SREBP-1 content increased also by using PKC inhibitors. These inhibitors increased the action of T3 on Akt phosphorylation suggesting that conventional PKCs may work as negative regulators of the T3-dependent SREBP-1 increase. T3 effects were partially abrogated by tetrac, an inhibitor of the T3-αvβ3 receptor interaction and partially evoked by T3 analog T3-agarose. These findings support a model in which T3 activates intracellular signaling pathways which may be involved in the increment of SREBP-1 level through an IRES-mediated translation mechanism. J. Cell. Physiol. 227: 2388-2397, 2012. © 2011 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Background and Purpose: We showed previously that a new Pt(II) complex ([Pt(O,O'-acac)(gamma-acac)(DMS)]) exerted high and fast apoptotic processes in MCF-7 cells. The objective of this study was to investigate the hypothesis that [Pt(O,O'-acac)(gamma-acac)(DMS)] is also able to exert anoikis and alter the migration ability of MCF-7 cells, and to show some of the signalling events leading to these alterations.Experimental Approach: Cells were treated with sublethal doses of [Pt(O,O'-acac)(gamma-acac)(DMS)], and the efficiency of colony initiation and anchorage-independent growth was assayed; cell migration was examined by in vitro culture wounding assay. Gelatin zymography for MMP-2 and -9 activities, Western blottings of MMPs, MAPKs, Src, PKC-epsilon and FAK, after [Pt(O,O'-acac)(gamma-acac)(DMS)] treatment, were also performed.Key Results: Sub-cytotoxic drug concentrations decreased the: (i) anchorage-dependent and -independent growth; (ii) migration ability; and (iii) expression and activity of MMP-2 and MMP-9. [Pt(O,O'-acac)(gamma-acac)(DMS)] provoked the generation of reactive oxygen species (ROS), and the activation of p38MAPK, Src and PKC-epsilon. p38MAPK phosphorylation, cell anoikis and migration due to [Pt(O,O'-acac)(gamma-acac)(DMS)] were blocked by PKC-epsilon inhibition. Furthermore, Src inhibition blocked the [Pt(O,O'-acac)(gamma-acac)(DMS)]-provoked activation of PKC-epsilon, while ROS generation blockage inhibited the activation of Src, and also the decrement of phosphorylated FAK observed in detached [Pt(O,O'-acac)(gamma-acac)(DMS)]-treated cells.Conclusions and Implications: Sublethal concentrations of [Pt(O,O'-acac)(gamma-acac)(DMS)] induced anoikis and prevented events leading to metastasis via alterations in cell migration, anchorage independency, stromal interactions and MMP activity. Hence, [Pt(O,O'-acac)(gamma-acac)(DMS)] may be a promising therapeutic agent for preventing growth and metastasis of breast cancer. [ABSTRACT FROM AUTHOR]

The article reports on the study that investigates the effectiveness of gene expression profiling in determining the prognosis of gliomatosis cerebi. It found that the well-known prognostic factor of gliomas are predictive of response or outcome in only a percentage of patients. Moreover, the results demonstrate a strong association between gene expression patterns and patient survival as well as a robust replicability of the gene expression-based predictors.

Background and Purpose: We showed previously that cisplatin inititates a signalling pathway mediated by PKC-delta/extracellular signal-regulated kinase (ERK), important for maintaining viability in PC Cl3 thyroid cells. The studies described herein examined whether c-fos was associated with cisplatin resistance and the signalling link between c-fos and PKC-delta/ERK.Experimental Approach: Cells were treated with various pharmacological inhibitors of PKCs and ERK, or were depleted of c-fos, PKC-delta, PKC-epsilon and caspase-3 by small interfering RNA (siRNA), then incubated with cisplatin and cytotoxicity assessed.Key Results: Cisplatin provokes the induction of c-fos and the activation of conventional PKC-beta, and novel PKC-delta and -epsilon. The cisplatin-provoked c-fos induction was decreased by Gö6976, a PKC-beta inhibitor; by siRNA for PKC-delta- but not that for PKC-epsilon or by PD98059, a mitogen-activated protein kinase/ERK kinase inhibitor. Expression of c-fos was abolished by GF109203X, an inhibitor of all PKC isoforms, or by PD98059 plus Gö6976 or by PKC-delta-siRNA plus Gö6976. When c-fos expression was blocked by siRNA, cisplatin cytotoxicity was strongly enhanced with increased caspase-3 activation. In PKC-delta-depleted cells treated with cisplatin, caspase-3 activation was increased and cell viability decreased. In these PKC-delta-depleted cells, PD98059 did not affect caspase-3 activation.Conclusions and Implications: In PC Cl3 cells, in the cell signalling pathways that lead to cisplatin resistance, PKC-delta controls ERK activity and, together with PKC-beta, also the induction of c-fos. Hence, the protective role of c-fos in thyroid cells has the potential to provide new opportunities for therapeutic intervention. [ABSTRACT FROM AUTHOR]

We demonstrated previously that in serum-starved MCF-7 breast cancer cell line, Ang II increased Na+/K+ATPase activity and activated the protein kinase C ζ (PKC-ζ) (Muscella et al., 2002 J Endocrinol 173:315–323; 2003 J Cell Physiol 197:61–68.). The aim of the present study was to investigate the modulation of the activity of the Na+/K+ATPase by PKC-ζ in MCF-7 cells. Here, using serum-starved MCF-7 cells, we have demonstrated that the effect of Ang II on the Na+/K+ATPase activity was inhibited by a synthetic myristoylated peptide with sequences based on the endogenous PKC-ζ pseudosubstrate region (ζ-PS) and by high doses of GF109203X, inhibitor of PKCs. When MCF-7 cells, grown in 10% fetal bovine serum (FBS), were stimulated with Ang II a dose- and time-dependent inhibition of the Na+/K+ATPase activity was obtained. Under this growth condition we found that mRNAs for AT1, AT2, and for Na+/K+ATPase α1 and α3 subunits were unchanged; besides both the activity of the Na+/K+ATPase and the level of PKC-ζ also were unaffected by the serum. The atypical PKC-ι level (present in very low abundance in serum-starved MCF-7) was increased and Ang II provoked its translocation from the cytosol to plasma membrane. PKC-ζ was localized to the membrane, and upon Ang II treatment its cellular localization did not change. The Ang II-mediated decrease of the Na+/K+ATPase activity was inhibited by high doses of GF109203X but not by ζ-PS, thus indicating that such effect was not due to PKC-ζ activity. The treatment of cells with PKC-ι antisense oligodeoxynucleotides inhibited the effects of Ang II on the Na+/K+ATPase activity. Additionally, the effect of Ang II on Na+/K+ATPase activity was also blocked by the phosphatidylinositol 3-kinase (PI3K) inhibitors, wortmannin and LY294002, and by the actin depolymerizing agents, cytochalasin D. In conclusion, in MCF-7 cells Ang II modulates the Na+/K+ATPase activity by both atypical PKC-ζ/-ι. The effects of Ang II are opposite depending upon the presence of the serum-sensitive PKC-ι, with the inhibitory effect possibly due to the redistribution of sodium pump from plasma membrane to the inactive intracellular pool. © 2005 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]