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The effect of silver nanoparticles (AgNPs) on proliferation and apoptosis of in ovo cultured glioblastoma multiforme (GBM) cells.

Urbańska K, Pająk B, Orzechowski A, Sokołowska J, Grodzik M, Sawosz E, Szmidt M, Sysa P - Nanoscale Res Lett (2015)

Bottom Line: The results show that AgNPs can influence GBM growth.Although there were statistically significant differences between control and AgNP groups in the AI and the levels of active caspase 9 and active caspase 3, the level of these proteins in GBM cells treated with AgNPs seems to be on the border between the spontaneous apoptosis and the induced.Our results indicate that the antiproliferative properties of silver nanoparticles overwhelm proapoptotic ones.

View Article: PubMed Central - PubMed

Affiliation: Division of Histology and Embryology, Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland.

ABSTRACT
Recently, it has been shown that silver nanoparticles (AgNPs) provide a unique approach to the treatment of tumors, especially those of neuroepithelial origin. Thus, the aim of this study was to evaluate the impact of AgNPs on proliferation and activation of the intrinsic apoptotic pathway of glioblastoma multiforme (GBM) cells cultured in an in ovo model. Human GBM cells, line U-87, were placed on chicken embryo chorioallantoic membrane. After 8 days, the tumors were divided into three groups: control (non-treated), treated with colloidal AgNPs (40 μg/ml), and placebo (tumors supplemented with vehicle only). At the end of the experiment, all tumors were isolated. Assessment of cell proliferation and cell apoptosis was estimated by histological, immunohistochemical, and Western blot analyses. The results show that AgNPs can influence GBM growth. AgNPs inhibit proliferation of GBM cells and seem to have proapoptotic properties. Although there were statistically significant differences between control and AgNP groups in the AI and the levels of active caspase 9 and active caspase 3, the level of these proteins in GBM cells treated with AgNPs seems to be on the border between the spontaneous apoptosis and the induced. Our results indicate that the antiproliferative properties of silver nanoparticles overwhelm proapoptotic ones. Further research focused on the cytotoxic effect of AgNPs on tumor and normal cells should be conducted.

No MeSH data available.


Related in: MedlinePlus

The level of active caspase 3 in GBM cells cultured in anin ovomodel. (A) Western blot analysis of the level of active caspase 3 in GBM cells from control group (C), placebo group (Pl), and AgNPs group (AgNPs). (B) Graph showing the optical density values (optical density, OD) obtained for the different bands, expressed in relative terms, defined as the ratio of OD values obtained for band representing active caspase 3 and OD values obtained for actin band. Letters (a, b) mean highly statistically significant differences, P ≤ 0.05. The obtained results are the mean of three independent experiments.
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Fig5: The level of active caspase 3 in GBM cells cultured in anin ovomodel. (A) Western blot analysis of the level of active caspase 3 in GBM cells from control group (C), placebo group (Pl), and AgNPs group (AgNPs). (B) Graph showing the optical density values (optical density, OD) obtained for the different bands, expressed in relative terms, defined as the ratio of OD values obtained for band representing active caspase 3 and OD values obtained for actin band. Letters (a, b) mean highly statistically significant differences, P ≤ 0.05. The obtained results are the mean of three independent experiments.

Mentions: The intensity of anti-active caspase 3 immunolabeling was stronger than with anti-caspase 9 antibody. Results obtained from immunochemical analysis of the presence of active caspase 3 were in agreement with those described for active caspase 9. In the control group, the percentage of active caspase 3+ cells (Figure 2G) was similar to the placebo group (Figure 2H) (2.80 to 11.30%, mean 5.65% ± 0.55%, and 3.40 to 11.30%, mean 6.07 ± 0.61, respectively). In the AgNPs group, casp3I was 5.50 to 19.50% (Figure 2H). The statistical analysis showed that the mean number of casp3I in the AgNPs group (10.46% ± 0.88%) was higher than in others (P ≤ 0.001), as shown in Figure 4. However, the lower limits of casp3I in AgNPs-treated tumors were similar to those in the control and placebo groups. When analyzed by Western blot, the level of active caspase 3 was shown to increase in AgNPs-treated tumors compared with other groups and the difference was statistically significant (P ≤ 0.05, Figure 5).Figure 5


The effect of silver nanoparticles (AgNPs) on proliferation and apoptosis of in ovo cultured glioblastoma multiforme (GBM) cells.

Urbańska K, Pająk B, Orzechowski A, Sokołowska J, Grodzik M, Sawosz E, Szmidt M, Sysa P - Nanoscale Res Lett (2015)

The level of active caspase 3 in GBM cells cultured in anin ovomodel. (A) Western blot analysis of the level of active caspase 3 in GBM cells from control group (C), placebo group (Pl), and AgNPs group (AgNPs). (B) Graph showing the optical density values (optical density, OD) obtained for the different bands, expressed in relative terms, defined as the ratio of OD values obtained for band representing active caspase 3 and OD values obtained for actin band. Letters (a, b) mean highly statistically significant differences, P ≤ 0.05. The obtained results are the mean of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4385140&req=5

Fig5: The level of active caspase 3 in GBM cells cultured in anin ovomodel. (A) Western blot analysis of the level of active caspase 3 in GBM cells from control group (C), placebo group (Pl), and AgNPs group (AgNPs). (B) Graph showing the optical density values (optical density, OD) obtained for the different bands, expressed in relative terms, defined as the ratio of OD values obtained for band representing active caspase 3 and OD values obtained for actin band. Letters (a, b) mean highly statistically significant differences, P ≤ 0.05. The obtained results are the mean of three independent experiments.
Mentions: The intensity of anti-active caspase 3 immunolabeling was stronger than with anti-caspase 9 antibody. Results obtained from immunochemical analysis of the presence of active caspase 3 were in agreement with those described for active caspase 9. In the control group, the percentage of active caspase 3+ cells (Figure 2G) was similar to the placebo group (Figure 2H) (2.80 to 11.30%, mean 5.65% ± 0.55%, and 3.40 to 11.30%, mean 6.07 ± 0.61, respectively). In the AgNPs group, casp3I was 5.50 to 19.50% (Figure 2H). The statistical analysis showed that the mean number of casp3I in the AgNPs group (10.46% ± 0.88%) was higher than in others (P ≤ 0.001), as shown in Figure 4. However, the lower limits of casp3I in AgNPs-treated tumors were similar to those in the control and placebo groups. When analyzed by Western blot, the level of active caspase 3 was shown to increase in AgNPs-treated tumors compared with other groups and the difference was statistically significant (P ≤ 0.05, Figure 5).Figure 5

Bottom Line: The results show that AgNPs can influence GBM growth.Although there were statistically significant differences between control and AgNP groups in the AI and the levels of active caspase 9 and active caspase 3, the level of these proteins in GBM cells treated with AgNPs seems to be on the border between the spontaneous apoptosis and the induced.Our results indicate that the antiproliferative properties of silver nanoparticles overwhelm proapoptotic ones.

View Article: PubMed Central - PubMed

Affiliation: Division of Histology and Embryology, Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland.

ABSTRACT
Recently, it has been shown that silver nanoparticles (AgNPs) provide a unique approach to the treatment of tumors, especially those of neuroepithelial origin. Thus, the aim of this study was to evaluate the impact of AgNPs on proliferation and activation of the intrinsic apoptotic pathway of glioblastoma multiforme (GBM) cells cultured in an in ovo model. Human GBM cells, line U-87, were placed on chicken embryo chorioallantoic membrane. After 8 days, the tumors were divided into three groups: control (non-treated), treated with colloidal AgNPs (40 μg/ml), and placebo (tumors supplemented with vehicle only). At the end of the experiment, all tumors were isolated. Assessment of cell proliferation and cell apoptosis was estimated by histological, immunohistochemical, and Western blot analyses. The results show that AgNPs can influence GBM growth. AgNPs inhibit proliferation of GBM cells and seem to have proapoptotic properties. Although there were statistically significant differences between control and AgNP groups in the AI and the levels of active caspase 9 and active caspase 3, the level of these proteins in GBM cells treated with AgNPs seems to be on the border between the spontaneous apoptosis and the induced. Our results indicate that the antiproliferative properties of silver nanoparticles overwhelm proapoptotic ones. Further research focused on the cytotoxic effect of AgNPs on tumor and normal cells should be conducted.

No MeSH data available.


Related in: MedlinePlus