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Increased Levels of NF-kB-Dependent Markers in Cancer-Associated Deep Venous Thrombosis.

Malaponte G, Signorelli SS, Bevelacqua V, Polesel J, Taborelli M, Guarneri C, Fenga C, Umezawa K, Libra M - PLoS ONE (2015)

Bottom Line: However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations.Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-.NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical and Biotechnological Sciences, Section of General & Clinical Pathology and Oncology, University of Catania, Catania, Italy.

ABSTRACT
Several studies highlight the role of inflammatory markers in thrombosis as well as in cancer. However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations. In the present study, C-reactive protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), matrix metalloproteases-9 (MMP-9), vascular endothelial growth factor (VEGF), tissue factor (TF), fibrinogen and soluble P-selectin, were analyzed in plasma and in monocyte samples from 385 cancer patients, of whom 64 were concomitantly affected by DVT (+). All these markers were higher in cancer patients DVT+ than in those DVT-. Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-. Significant correlation between data obtained in plasma and monocyte samples was observed. NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-. To further demonstrate the involvement of NF-kB activation by the above mentioned molecules, we treated monocyte derived from healthy donors with a pool of sera from cancer patients with and without DVT. These set of experiments further suggest the significant role played by some molecules, regulated by NF-kB, and detected in cancer patients with DVT. Our data support the notion that NF-kB may be considered as a therapeutic target for cancer patients, especially those complicated by DVT. Treatment with NF-kB inhibitors may represent a possible strategy to prevent or reduce the risk of DVT in cancer patients.

No MeSH data available.


Related in: MedlinePlus

Serum-dependent activation of nuclear factor (NF)–NF-kB p65 subunit in healthy monocytes.Monocytes from 25 healthy controls were evaluated for NF-kB activation after they were cultured for 20 hours in medium supplemented with either 40% serum from three groups. Monocytes (5x105) from 25 healthy donors were cultured for 20 hours in medium (RPMI 1640) supplemented with either 40% serum derived from 64 cancer patients DVT+ and 257 DVT- with the highest cytokines plasma levels (> 75th percentile) or 40% serum derived from 100 healthy donors with the lowest cytokines values (< 25th percentile). The incubation of healthy monocytes with pooled sera derived from cancer patients DVT+ (sCADVT+) or DVT- (sCADVT-) induced a significant increase of NF-kB activity compared with that derived from healthy controls (HM) after treatment with sera from healthy controls (SH) (P<0.0001). An higher NF-kB p65 subunit activation was observed in monocytes stimulated by sera from cancer patients DVT+ compared to that stimulated by sera derived from DVT- (P<0.001) (t-test). No NF-kB p65 subunit activation was observed in monocytes stimulated with sera derived from healthy controls.
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pone.0132496.g005: Serum-dependent activation of nuclear factor (NF)–NF-kB p65 subunit in healthy monocytes.Monocytes from 25 healthy controls were evaluated for NF-kB activation after they were cultured for 20 hours in medium supplemented with either 40% serum from three groups. Monocytes (5x105) from 25 healthy donors were cultured for 20 hours in medium (RPMI 1640) supplemented with either 40% serum derived from 64 cancer patients DVT+ and 257 DVT- with the highest cytokines plasma levels (> 75th percentile) or 40% serum derived from 100 healthy donors with the lowest cytokines values (< 25th percentile). The incubation of healthy monocytes with pooled sera derived from cancer patients DVT+ (sCADVT+) or DVT- (sCADVT-) induced a significant increase of NF-kB activity compared with that derived from healthy controls (HM) after treatment with sera from healthy controls (SH) (P<0.0001). An higher NF-kB p65 subunit activation was observed in monocytes stimulated by sera from cancer patients DVT+ compared to that stimulated by sera derived from DVT- (P<0.001) (t-test). No NF-kB p65 subunit activation was observed in monocytes stimulated with sera derived from healthy controls.

Mentions: As shown in Fig 5, the incubation of healthy monocytes with pooled sera, derived from the cancer patients DVT+ or DVT- with the highest values of cytokines, induced a significant increase of NF-kB activation compared with pooled sera, derived from the healthy controls with the lowest values of the same cytokines, (p<0.0001). In addition, a higher NF-kB activation was observed in monocytes stimulated by sera derived from cancer patients DVT+ compared to that stimulated by sera derived from DVT- (p<0.001) (Fig 5). In order to further exclude the possibility that some LPS contamination might contribute to the NF-kB activation, 10 μg/ml of polymixin B was added in all cell cultures to neutralize any potential LPS contamination. Instead, as positive control of NF-kB activation, LPS was considered. LPS-stimulated cultures induced an higher NF-kB activity statistically significant compared to cancer derived sera-stimulated cultures (p<0.001), while no significant difference was evident between LPS-stimulated cultures and DVT cancer derived sera- stimulated cultures. In the LPS experiments no polymixin B was added. The NF-kB activation, stimulated with the sera derived from cancer patients with and without DVT or LPS, was partially blocked by DHMEQ (10 μg/mL) when added in both cultures and LPS-stimulated cultures.


Increased Levels of NF-kB-Dependent Markers in Cancer-Associated Deep Venous Thrombosis.

Malaponte G, Signorelli SS, Bevelacqua V, Polesel J, Taborelli M, Guarneri C, Fenga C, Umezawa K, Libra M - PLoS ONE (2015)

Serum-dependent activation of nuclear factor (NF)–NF-kB p65 subunit in healthy monocytes.Monocytes from 25 healthy controls were evaluated for NF-kB activation after they were cultured for 20 hours in medium supplemented with either 40% serum from three groups. Monocytes (5x105) from 25 healthy donors were cultured for 20 hours in medium (RPMI 1640) supplemented with either 40% serum derived from 64 cancer patients DVT+ and 257 DVT- with the highest cytokines plasma levels (> 75th percentile) or 40% serum derived from 100 healthy donors with the lowest cytokines values (< 25th percentile). The incubation of healthy monocytes with pooled sera derived from cancer patients DVT+ (sCADVT+) or DVT- (sCADVT-) induced a significant increase of NF-kB activity compared with that derived from healthy controls (HM) after treatment with sera from healthy controls (SH) (P<0.0001). An higher NF-kB p65 subunit activation was observed in monocytes stimulated by sera from cancer patients DVT+ compared to that stimulated by sera derived from DVT- (P<0.001) (t-test). No NF-kB p65 subunit activation was observed in monocytes stimulated with sera derived from healthy controls.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4507873&req=5

pone.0132496.g005: Serum-dependent activation of nuclear factor (NF)–NF-kB p65 subunit in healthy monocytes.Monocytes from 25 healthy controls were evaluated for NF-kB activation after they were cultured for 20 hours in medium supplemented with either 40% serum from three groups. Monocytes (5x105) from 25 healthy donors were cultured for 20 hours in medium (RPMI 1640) supplemented with either 40% serum derived from 64 cancer patients DVT+ and 257 DVT- with the highest cytokines plasma levels (> 75th percentile) or 40% serum derived from 100 healthy donors with the lowest cytokines values (< 25th percentile). The incubation of healthy monocytes with pooled sera derived from cancer patients DVT+ (sCADVT+) or DVT- (sCADVT-) induced a significant increase of NF-kB activity compared with that derived from healthy controls (HM) after treatment with sera from healthy controls (SH) (P<0.0001). An higher NF-kB p65 subunit activation was observed in monocytes stimulated by sera from cancer patients DVT+ compared to that stimulated by sera derived from DVT- (P<0.001) (t-test). No NF-kB p65 subunit activation was observed in monocytes stimulated with sera derived from healthy controls.
Mentions: As shown in Fig 5, the incubation of healthy monocytes with pooled sera, derived from the cancer patients DVT+ or DVT- with the highest values of cytokines, induced a significant increase of NF-kB activation compared with pooled sera, derived from the healthy controls with the lowest values of the same cytokines, (p<0.0001). In addition, a higher NF-kB activation was observed in monocytes stimulated by sera derived from cancer patients DVT+ compared to that stimulated by sera derived from DVT- (p<0.001) (Fig 5). In order to further exclude the possibility that some LPS contamination might contribute to the NF-kB activation, 10 μg/ml of polymixin B was added in all cell cultures to neutralize any potential LPS contamination. Instead, as positive control of NF-kB activation, LPS was considered. LPS-stimulated cultures induced an higher NF-kB activity statistically significant compared to cancer derived sera-stimulated cultures (p<0.001), while no significant difference was evident between LPS-stimulated cultures and DVT cancer derived sera- stimulated cultures. In the LPS experiments no polymixin B was added. The NF-kB activation, stimulated with the sera derived from cancer patients with and without DVT or LPS, was partially blocked by DHMEQ (10 μg/mL) when added in both cultures and LPS-stimulated cultures.

Bottom Line: However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations.Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-.NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical and Biotechnological Sciences, Section of General & Clinical Pathology and Oncology, University of Catania, Catania, Italy.

ABSTRACT
Several studies highlight the role of inflammatory markers in thrombosis as well as in cancer. However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations. In the present study, C-reactive protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), matrix metalloproteases-9 (MMP-9), vascular endothelial growth factor (VEGF), tissue factor (TF), fibrinogen and soluble P-selectin, were analyzed in plasma and in monocyte samples from 385 cancer patients, of whom 64 were concomitantly affected by DVT (+). All these markers were higher in cancer patients DVT+ than in those DVT-. Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-. Significant correlation between data obtained in plasma and monocyte samples was observed. NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-. To further demonstrate the involvement of NF-kB activation by the above mentioned molecules, we treated monocyte derived from healthy donors with a pool of sera from cancer patients with and without DVT. These set of experiments further suggest the significant role played by some molecules, regulated by NF-kB, and detected in cancer patients with DVT. Our data support the notion that NF-kB may be considered as a therapeutic target for cancer patients, especially those complicated by DVT. Treatment with NF-kB inhibitors may represent a possible strategy to prevent or reduce the risk of DVT in cancer patients.

No MeSH data available.


Related in: MedlinePlus