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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

Diagrams presenting quantitative determination of platinum(II) content in DNA and proteins in HeLa cells, obtained by ICP-OES analysis, following 6 h and 24 h of action of 1, 2 or CDDP; A Platinum content in cellular DNA; B Platinum content in cellular protein fraction (pg Pt/mg proteins). Bar graph represent mean ± SD of three independent experiments.
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f5-rado-47-04-346: Diagrams presenting quantitative determination of platinum(II) content in DNA and proteins in HeLa cells, obtained by ICP-OES analysis, following 6 h and 24 h of action of 1, 2 or CDDP; A Platinum content in cellular DNA; B Platinum content in cellular protein fraction (pg Pt/mg proteins). Bar graph represent mean ± SD of three independent experiments.

Mentions: Intracellular platinum(II) distribution among DNA and protein fractions in HeLa cells treated with equitoxic concentrations of investigated complexes for 6 and 24 h, was analyzed using ICP-OES analysis, and results are presented in Figure 5. Levels of platinum(II)-DNA binding (Figure 5A), varied between the investigated complexes, especially following short-term (6 h) treatment, when platinum content (pg Pt/mg DNA) decreased in order: 21 ± 2.5 (complex 1); 14.5 ±0.4 (CDDP) and 6.4±1.1 (complex 2). Both CDDP and 1 seemed to be more effcient in promoting cellular DNA binding comparing to complex 2, though differences in double-stranded DNA platination affinity between complex 2 and CDDP were in accordance to the recent study.34 Platinum(II)-DNA content, decreased in time-dependent manner, and reached comparable levels following 24 h of action. Results of the ICPOES analysis of platinum(II) content in the protein fraction (Figure 5B), indicated that 1 exhibited the highest affinity for protein binding following both 6 h and 24 h treatment, while 2 exhibited the lowest binding affinity. Time dependent decrease of protein binding, indicated reversible nature of interactions of 1 and 2, oppositely to CDDP.


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)

Diagrams presenting quantitative determination of platinum(II) content in DNA and proteins in HeLa cells, obtained by ICP-OES analysis, following 6 h and 24 h of action of 1, 2 or CDDP; A Platinum content in cellular DNA; B Platinum content in cellular protein fraction (pg Pt/mg proteins). Bar graph represent mean ± 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

f5-rado-47-04-346: Diagrams presenting quantitative determination of platinum(II) content in DNA and proteins in HeLa cells, obtained by ICP-OES analysis, following 6 h and 24 h of action of 1, 2 or CDDP; A Platinum content in cellular DNA; B Platinum content in cellular protein fraction (pg Pt/mg proteins). Bar graph represent mean ± SD of three independent experiments.
Mentions: Intracellular platinum(II) distribution among DNA and protein fractions in HeLa cells treated with equitoxic concentrations of investigated complexes for 6 and 24 h, was analyzed using ICP-OES analysis, and results are presented in Figure 5. Levels of platinum(II)-DNA binding (Figure 5A), varied between the investigated complexes, especially following short-term (6 h) treatment, when platinum content (pg Pt/mg DNA) decreased in order: 21 ± 2.5 (complex 1); 14.5 ±0.4 (CDDP) and 6.4±1.1 (complex 2). Both CDDP and 1 seemed to be more effcient in promoting cellular DNA binding comparing to complex 2, though differences in double-stranded DNA platination affinity between complex 2 and CDDP were in accordance to the recent study.34 Platinum(II)-DNA content, decreased in time-dependent manner, and reached comparable levels following 24 h of action. Results of the ICPOES analysis of platinum(II) content in the protein fraction (Figure 5B), indicated that 1 exhibited the highest affinity for protein binding following both 6 h and 24 h treatment, while 2 exhibited the lowest binding affinity. Time dependent decrease of protein binding, indicated reversible nature of interactions of 1 and 2, oppositely to CDDP.

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