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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 Dot plot diagrams obtained by flow-cytometric analysis of treated HeLa cells after dual staining with Annexin V-FITC and PI. Annexin V-FITC/PI staining was monitored overtime, following 4 and 24 hours in HeLa cells exposed to complex 1, 2 or CDDP at concentrations corresponding to IC50. Representative dot plots of three independent experiments are given, presenting intact cells at lower-left quadrant, FITC(-)/PI(-); early apoptotic cells at lower-right quadrant, FITC(+)/PI(-); late apoptotic or necrotic cells at upper-right quadrant, FITC(+)/PI(+); and necrotic cells at upper-left quadrant, FITC(-)/PI(+). B Apoptosis and necrosis were quanitified by FACS after Annexin V-FITC and PI labeling; bar graphs represent mean ± SD in at least three independent experiments.
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f4-rado-47-04-346: A Dot plot diagrams obtained by flow-cytometric analysis of treated HeLa cells after dual staining with Annexin V-FITC and PI. Annexin V-FITC/PI staining was monitored overtime, following 4 and 24 hours in HeLa cells exposed to complex 1, 2 or CDDP at concentrations corresponding to IC50. Representative dot plots of three independent experiments are given, presenting intact cells at lower-left quadrant, FITC(-)/PI(-); early apoptotic cells at lower-right quadrant, FITC(+)/PI(-); late apoptotic or necrotic cells at upper-right quadrant, FITC(+)/PI(+); and necrotic cells at upper-left quadrant, FITC(-)/PI(+). B Apoptosis and necrosis were quanitified by FACS after Annexin V-FITC and PI labeling; bar graphs represent mean ± SD in at least three independent experiments.

Mentions: Potential of the investigated complex to induce apoptosis in HeLa cells was assessed by flow cytometry using Annexin V-FITC and PI dual staining. Dot plots are presented in Figure 4A, while Figure 4B reports the results of a representative experiment as percentages of apoptotic cells (Annexin V-FITC positive and PI negative) and necrotic cells (Annexin V-FITC negative and PI positive) measured periodically at 4 and 24 h. Data obtained indicated that complex 2 caused 15.5% of apoptosis in HeLa cells following 24 h of action, while the percentage of necrotic cells was negligible. Kinetics, as well as the degree of apoptosis induction, was comparable to CDDP. Complex 1 at concentration of IC50 initiated early apoptotic cell death after 4 and 24 h action, where apoptotic cell population represented 14.2% and 15.5% of total cells, respectively. Though, after 24 h of action more than 50% of total cell population underwent cell death in either apoptotic or necrotic manner.


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 Dot plot diagrams obtained by flow-cytometric analysis of treated HeLa cells after dual staining with Annexin V-FITC and PI. Annexin V-FITC/PI staining was monitored overtime, following 4 and 24 hours in HeLa cells exposed to complex 1, 2 or CDDP at concentrations corresponding to IC50. Representative dot plots of three independent experiments are given, presenting intact cells at lower-left quadrant, FITC(-)/PI(-); early apoptotic cells at lower-right quadrant, FITC(+)/PI(-); late apoptotic or necrotic cells at upper-right quadrant, FITC(+)/PI(+); and necrotic cells at upper-left quadrant, FITC(-)/PI(+). B Apoptosis and necrosis were quanitified by FACS after Annexin V-FITC and PI labeling; bar graphs represent mean ± SD in at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4-rado-47-04-346: A Dot plot diagrams obtained by flow-cytometric analysis of treated HeLa cells after dual staining with Annexin V-FITC and PI. Annexin V-FITC/PI staining was monitored overtime, following 4 and 24 hours in HeLa cells exposed to complex 1, 2 or CDDP at concentrations corresponding to IC50. Representative dot plots of three independent experiments are given, presenting intact cells at lower-left quadrant, FITC(-)/PI(-); early apoptotic cells at lower-right quadrant, FITC(+)/PI(-); late apoptotic or necrotic cells at upper-right quadrant, FITC(+)/PI(+); and necrotic cells at upper-left quadrant, FITC(-)/PI(+). B Apoptosis and necrosis were quanitified by FACS after Annexin V-FITC and PI labeling; bar graphs represent mean ± SD in at least three independent experiments.
Mentions: Potential of the investigated complex to induce apoptosis in HeLa cells was assessed by flow cytometry using Annexin V-FITC and PI dual staining. Dot plots are presented in Figure 4A, while Figure 4B reports the results of a representative experiment as percentages of apoptotic cells (Annexin V-FITC positive and PI negative) and necrotic cells (Annexin V-FITC negative and PI positive) measured periodically at 4 and 24 h. Data obtained indicated that complex 2 caused 15.5% of apoptosis in HeLa cells following 24 h of action, while the percentage of necrotic cells was negligible. Kinetics, as well as the degree of apoptosis induction, was comparable to CDDP. Complex 1 at concentration of IC50 initiated early apoptotic cell death after 4 and 24 h action, where apoptotic cell population represented 14.2% and 15.5% of total cells, respectively. Though, after 24 h of action more than 50% of total cell population underwent cell death in either apoptotic or necrotic manner.

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