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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 cell cycle phase distribution of treated HeLa cells, obtained by flow-cytometric analysis of the DNA content in fixed cells, after staining with PI. HeLa cells were collected following 24 h treatment with tested complexes or cisplatin at concentration corresponding to A IC50 and B 1.5×IC50. Bar graphs represent mean ± SD in at least three independent experiments. Asteriks (*) denotes p values < 0.05, calculated by Student t-test, indicating statisticaly significant differences (Stata Software).
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f3-rado-47-04-346: Diagrams presenting cell cycle phase distribution of treated HeLa cells, obtained by flow-cytometric analysis of the DNA content in fixed cells, after staining with PI. HeLa cells were collected following 24 h treatment with tested complexes or cisplatin at concentration corresponding to A IC50 and B 1.5×IC50. Bar graphs represent mean ± SD in at least three independent experiments. Asteriks (*) denotes p values < 0.05, calculated by Student t-test, indicating statisticaly significant differences (Stata Software).

Mentions: The potential of the tested complexes to induce cell cycle alterations in comparison to CDDP in HeLa cells, was examined by flow cytometry using staining with PI. Results are presented as diagrams of cell distribution over the cell cycle phases after 24 h of agent action, where Figure 3A shows effects of the complexes at concentration corresponding to IC50, and Figure 3B shows effects of the complexes at concentration corresponding to 1.5×IC50. Complex 1, induced arrest in the S phase of cell cycle at concentrations corresponding to 1.5 × IC50, but less than CDDP (Figure 3B). Complex 1 induced decrease of cell percentage in the G0/G1 phase in concentration dependent manner, comparing to the non-treated control. CDDP induced dose-dependent arrest in the S phase of cell cycle, and decrease of cell progression through G2/M phase (Figures 3A and 3B).


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 cell cycle phase distribution of treated HeLa cells, obtained by flow-cytometric analysis of the DNA content in fixed cells, after staining with PI. HeLa cells were collected following 24 h treatment with tested complexes or cisplatin at concentration corresponding to A IC50 and B 1.5×IC50. Bar graphs represent mean ± SD in at least three independent experiments. Asteriks (*) denotes p values < 0.05, calculated by Student t-test, indicating statisticaly significant differences (Stata Software).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-rado-47-04-346: Diagrams presenting cell cycle phase distribution of treated HeLa cells, obtained by flow-cytometric analysis of the DNA content in fixed cells, after staining with PI. HeLa cells were collected following 24 h treatment with tested complexes or cisplatin at concentration corresponding to A IC50 and B 1.5×IC50. Bar graphs represent mean ± SD in at least three independent experiments. Asteriks (*) denotes p values < 0.05, calculated by Student t-test, indicating statisticaly significant differences (Stata Software).
Mentions: The potential of the tested complexes to induce cell cycle alterations in comparison to CDDP in HeLa cells, was examined by flow cytometry using staining with PI. Results are presented as diagrams of cell distribution over the cell cycle phases after 24 h of agent action, where Figure 3A shows effects of the complexes at concentration corresponding to IC50, and Figure 3B shows effects of the complexes at concentration corresponding to 1.5×IC50. Complex 1, induced arrest in the S phase of cell cycle at concentrations corresponding to 1.5 × IC50, but less than CDDP (Figure 3B). Complex 1 induced decrease of cell percentage in the G0/G1 phase in concentration dependent manner, comparing to the non-treated control. CDDP induced dose-dependent arrest in the S phase of cell cycle, and decrease of cell progression through G2/M phase (Figures 3A and 3B).

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