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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 Results of the qRT-PCR analysis of ERCC1 mRNA presented as diagrams showing relative expression level of ERCC1 mRNA, normalized with the GAPDH; Bar graph represent mean ± SD of three independent experiments; B Protein expression levels of ERCC1 determined by Western blot, and normalized with b-actin. Tested agents 1, 2 and CDDP were applied at concentration of 0.5×IC50. Western blot results show one representative experiment selected of three.
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f6-rado-47-04-346: A Results of the qRT-PCR analysis of ERCC1 mRNA presented as diagrams showing relative expression level of ERCC1 mRNA, normalized with the GAPDH; Bar graph represent mean ± SD of three independent experiments; B Protein expression levels of ERCC1 determined by Western blot, and normalized with b-actin. Tested agents 1, 2 and CDDP were applied at concentration of 0.5×IC50. Western blot results show one representative experiment selected of three.

Mentions: DNA excision repair protein ERCC1 is an important component of NER (Nucleotide Excision Repair) which is primarily induced in the repair of bulky platinum-DNA adducts.35 In order to evaluate whether investigated complexes induce ERCC1-dependent cell response as the result of cytotoxic DNA lesions, we investigated mRNA and protein expression level of ERCC1. Data obtained on HeLa cells after 6 h of continuous treatment with equitoxic concentrations of tested trans-platinum complexes or CDDP indicated negative modulation of ERCC1 expression on both mRNA and protein levels (results presented in Figure 6). Complexes 1, 2 and CDDP decreased ERCC1 mRNA level for 45%, 40% and 36%, respectively, comparing to the non treated control (Figure 6A). Western blot analysis (Figure 6B) showed reduction of ERCC1 protein levels, following both trans-complexes 1 and 2 action, while there were no obvious changes associated with CDDP treatment.


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 Results of the qRT-PCR analysis of ERCC1 mRNA presented as diagrams showing relative expression level of ERCC1 mRNA, normalized with the GAPDH; Bar graph represent mean ± SD of three independent experiments; B Protein expression levels of ERCC1 determined by Western blot, and normalized with b-actin. Tested agents 1, 2 and CDDP were applied at concentration of 0.5×IC50. Western blot results show one representative experiment selected of three.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6-rado-47-04-346: A Results of the qRT-PCR analysis of ERCC1 mRNA presented as diagrams showing relative expression level of ERCC1 mRNA, normalized with the GAPDH; Bar graph represent mean ± SD of three independent experiments; B Protein expression levels of ERCC1 determined by Western blot, and normalized with b-actin. Tested agents 1, 2 and CDDP were applied at concentration of 0.5×IC50. Western blot results show one representative experiment selected of three.
Mentions: DNA excision repair protein ERCC1 is an important component of NER (Nucleotide Excision Repair) which is primarily induced in the repair of bulky platinum-DNA adducts.35 In order to evaluate whether investigated complexes induce ERCC1-dependent cell response as the result of cytotoxic DNA lesions, we investigated mRNA and protein expression level of ERCC1. Data obtained on HeLa cells after 6 h of continuous treatment with equitoxic concentrations of tested trans-platinum complexes or CDDP indicated negative modulation of ERCC1 expression on both mRNA and protein levels (results presented in Figure 6). Complexes 1, 2 and CDDP decreased ERCC1 mRNA level for 45%, 40% and 36%, respectively, comparing to the non treated control (Figure 6A). Western blot analysis (Figure 6B) showed reduction of ERCC1 protein levels, following both trans-complexes 1 and 2 action, while there were no obvious changes associated with CDDP treatment.

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