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Biological evaluation of transdichloridoplatinum(II) complexes with 3- and 4-acetylpyridine in comparison to cisplatin.

Filipovic L, Arandelovic S, Gligorijevic N, Krivokuca A, Jankovic R, Srdic-Rajic T, Rakic G, Tesic Z, Radulovic S - Radiol Oncol (2013)

Bottom Line: Compound 2 displayed an apparent cytoselective profile, and flow cytometry analysis in HeLa cells indicated that 2 exerted antiproliferative effect through apoptosis induction, while 1 induced both apoptosis and necrosis.Action of 1 and 2, as analyzed by quantitative real-time PCR and Western blot, was associated with down-regulation of ERCC1.Both trans-complexes inhibited MMP-9 mRNA expression in HeLa, while 2 significantly abrogated in vitro tubulogenesis in MS1 cells.

View Article: PubMed Central - PubMed

Affiliation: Institute for Oncology and Radiology of Serbia, Belgrade, Serbia.

ABSTRACT

Background: In our previous study we reported the synthesis and cytotoxicity of two trans-platinum(II) complexes: trans-[PtCl2(3-acetylpyridine)2] (1) and trans-[PtCl2(4-acetylpyridine)2] (2), revealing significant cytotoxic potential of 2. In order to evaluate the mechanism underlying biological activity of both trans-Pt(II) isomers, comparative studies versus cisplatin were performed in HeLa, MRC-5 and MS1 cells.

Materials and methods: The cytotoxic activity of the investigated complexes was determined using SRB assay. The colagenolytic activity was determined using gelatin zymography, while the effect of platinum complexes on matrix metalloproteinases 2 and 9 mRNA expression was evaluated by quantitative real-time PCR. Apoptotic potential and cell cycle alterations were determined by FACS analyses. Western blot analysis was used to evaluate the effect on expression of DNA-repair enzyme ERCC1, and quantitative real-time PCR was used for the ERCC1 mRNA expression analysis. In vitro antiangiogenic potential was determined by tube formation assay. Platinum content in intracellular DNA and proteins was determined by inductively coupled plasma-optical emission spectrometry.

Results: Compound 2 displayed an apparent cytoselective profile, and flow cytometry analysis in HeLa cells indicated that 2 exerted antiproliferative effect through apoptosis induction, while 1 induced both apoptosis and necrosis. Action of 1 and 2, as analyzed by quantitative real-time PCR and Western blot, was associated with down-regulation of ERCC1. Both trans-complexes inhibited MMP-9 mRNA expression in HeLa, while 2 significantly abrogated in vitro tubulogenesis in MS1 cells.

Conclusions: The ability of 2 to induce multiple and selective in vitro cytotoxic effects encourages further investigations of trans-platinum(II) complexes with substituted pyridines.

No MeSH data available.


Related in: MedlinePlus

A Micrographs of tube formation of MS1 cells, taken following 24 h treatment with sub-toxic concentrations (0.03×IC50) of complexes 1, 2 or CDDP, or without treatment (control cells); B Results of the gelatin zymography following 6 h treatment with 0.5×IC50 of 1, 2 or CDDP. Supernatants were collected and equal amounts of proteins were analysed by zymography. Representative of three independent experiments; C Diagram presenting quantification of catalytic activity of secreted MMP-2 and MMP-9 in treated HeLa cells, obtained by quantitative analysis of zymograms, using Image J software. Data are expressed in arbitrary units as mean ± standard deviation of three independent experiments; D Results of the qRT-PCR analysis of the MMP-2 and MMP-9 mRNA expression level in HeLa cells after 6 h treatment with 1, 2 or CDDP, at concentration 0.5×IC50. Each bar represents the average (±SD) of three independent experiments.
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f7-rado-47-04-346: A Micrographs of tube formation of MS1 cells, taken following 24 h treatment with sub-toxic concentrations (0.03×IC50) of complexes 1, 2 or CDDP, or without treatment (control cells); B Results of the gelatin zymography following 6 h treatment with 0.5×IC50 of 1, 2 or CDDP. Supernatants were collected and equal amounts of proteins were analysed by zymography. Representative of three independent experiments; C Diagram presenting quantification of catalytic activity of secreted MMP-2 and MMP-9 in treated HeLa cells, obtained by quantitative analysis of zymograms, using Image J software. Data are expressed in arbitrary units as mean ± standard deviation of three independent experiments; D Results of the qRT-PCR analysis of the MMP-2 and MMP-9 mRNA expression level in HeLa cells after 6 h treatment with 1, 2 or CDDP, at concentration 0.5×IC50. Each bar represents the average (±SD) of three independent experiments.

Mentions: In order to determine the potency of the investigated complexes to restrict the angiogenesis of cancer cells, we performed an in vitro tube formation assay in mouse endothelial cells MS1. In our experiment, MS1 endothelial cells were treated with sub-toxic concentrations of the investigated complexes in order to distinguish among growth inhibitory effect and their potential to inhibit the formation of tube-like structures. Antiangiogenic effect was observed for both tested trans-platinum complexes, and results are presented in Figure 7A. Trans-complexes, particularly complex 2, showed inhibitory effect on the formation of cell-cell contact and tube-like structures, at very low sub-toxic concentration corresponding to 0.03×IC50, while CDDP did not exhibit any significant effect in this assay.


Biological evaluation of transdichloridoplatinum(II) complexes with 3- and 4-acetylpyridine in comparison to cisplatin.

Filipovic L, Arandelovic S, Gligorijevic N, Krivokuca A, Jankovic R, Srdic-Rajic T, Rakic G, Tesic Z, Radulovic S - Radiol Oncol (2013)

A Micrographs of tube formation of MS1 cells, taken following 24 h treatment with sub-toxic concentrations (0.03×IC50) of complexes 1, 2 or CDDP, or without treatment (control cells); B Results of the gelatin zymography following 6 h treatment with 0.5×IC50 of 1, 2 or CDDP. Supernatants were collected and equal amounts of proteins were analysed by zymography. Representative of three independent experiments; C Diagram presenting quantification of catalytic activity of secreted MMP-2 and MMP-9 in treated HeLa cells, obtained by quantitative analysis of zymograms, using Image J software. Data are expressed in arbitrary units as mean ± standard deviation of three independent experiments; D Results of the qRT-PCR analysis of the MMP-2 and MMP-9 mRNA expression level in HeLa cells after 6 h treatment with 1, 2 or CDDP, at concentration 0.5×IC50. Each bar represents the average (±SD) of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3814279&req=5

f7-rado-47-04-346: A Micrographs of tube formation of MS1 cells, taken following 24 h treatment with sub-toxic concentrations (0.03×IC50) of complexes 1, 2 or CDDP, or without treatment (control cells); B Results of the gelatin zymography following 6 h treatment with 0.5×IC50 of 1, 2 or CDDP. Supernatants were collected and equal amounts of proteins were analysed by zymography. Representative of three independent experiments; C Diagram presenting quantification of catalytic activity of secreted MMP-2 and MMP-9 in treated HeLa cells, obtained by quantitative analysis of zymograms, using Image J software. Data are expressed in arbitrary units as mean ± standard deviation of three independent experiments; D Results of the qRT-PCR analysis of the MMP-2 and MMP-9 mRNA expression level in HeLa cells after 6 h treatment with 1, 2 or CDDP, at concentration 0.5×IC50. Each bar represents the average (±SD) of three independent experiments.
Mentions: In order to determine the potency of the investigated complexes to restrict the angiogenesis of cancer cells, we performed an in vitro tube formation assay in mouse endothelial cells MS1. In our experiment, MS1 endothelial cells were treated with sub-toxic concentrations of the investigated complexes in order to distinguish among growth inhibitory effect and their potential to inhibit the formation of tube-like structures. Antiangiogenic effect was observed for both tested trans-platinum complexes, and results are presented in Figure 7A. Trans-complexes, particularly complex 2, showed inhibitory effect on the formation of cell-cell contact and tube-like structures, at very low sub-toxic concentration corresponding to 0.03×IC50, while CDDP did not exhibit any significant effect in this assay.

Bottom Line: Compound 2 displayed an apparent cytoselective profile, and flow cytometry analysis in HeLa cells indicated that 2 exerted antiproliferative effect through apoptosis induction, while 1 induced both apoptosis and necrosis.Action of 1 and 2, as analyzed by quantitative real-time PCR and Western blot, was associated with down-regulation of ERCC1.Both trans-complexes inhibited MMP-9 mRNA expression in HeLa, while 2 significantly abrogated in vitro tubulogenesis in MS1 cells.

View Article: PubMed Central - PubMed

Affiliation: Institute for Oncology and Radiology of Serbia, Belgrade, Serbia.

ABSTRACT

Background: In our previous study we reported the synthesis and cytotoxicity of two trans-platinum(II) complexes: trans-[PtCl2(3-acetylpyridine)2] (1) and trans-[PtCl2(4-acetylpyridine)2] (2), revealing significant cytotoxic potential of 2. In order to evaluate the mechanism underlying biological activity of both trans-Pt(II) isomers, comparative studies versus cisplatin were performed in HeLa, MRC-5 and MS1 cells.

Materials and methods: The cytotoxic activity of the investigated complexes was determined using SRB assay. The colagenolytic activity was determined using gelatin zymography, while the effect of platinum complexes on matrix metalloproteinases 2 and 9 mRNA expression was evaluated by quantitative real-time PCR. Apoptotic potential and cell cycle alterations were determined by FACS analyses. Western blot analysis was used to evaluate the effect on expression of DNA-repair enzyme ERCC1, and quantitative real-time PCR was used for the ERCC1 mRNA expression analysis. In vitro antiangiogenic potential was determined by tube formation assay. Platinum content in intracellular DNA and proteins was determined by inductively coupled plasma-optical emission spectrometry.

Results: Compound 2 displayed an apparent cytoselective profile, and flow cytometry analysis in HeLa cells indicated that 2 exerted antiproliferative effect through apoptosis induction, while 1 induced both apoptosis and necrosis. Action of 1 and 2, as analyzed by quantitative real-time PCR and Western blot, was associated with down-regulation of ERCC1. Both trans-complexes inhibited MMP-9 mRNA expression in HeLa, while 2 significantly abrogated in vitro tubulogenesis in MS1 cells.

Conclusions: The ability of 2 to induce multiple and selective in vitro cytotoxic effects encourages further investigations of trans-platinum(II) complexes with substituted pyridines.

No MeSH data available.


Related in: MedlinePlus