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Novel piperazine core compound induces death in human liver cancer cells: possible pharmacological properties.

Samie N, Muniandy S, Kanthimathi MS, Haerian BS, Azudin RE - Sci Rep (2016)

Bottom Line: PCC displayed a strong suppressive effect on SNU-475 and SNU-423 cells with an IC50 value of 6.98 ± 0.11 μg/ml and 7.76 ± 0.45 μg/ml respectively, after 24 h of treatment.Activation of this pathway was further evidenced by significant activation of caspase 3/7 and 9.Cell cycle arrest in the G1 phase was confirmed by flow cytometry and up-regulation of glutathione reductase expression was quantified by qPCR.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
The current study evaluates the cytotoxic mechanism of a novel piperazine derivate designated as PCC against human liver cancer cells. In this context, human liver cancer cell lines, SNU-475 and 243, human monocyte/macrophage cell line, CRL-9855, and human B lymphocyte cell line, CCL-156, were used to determine the IC50 of PCC using the standard MTT assay. PCC displayed a strong suppressive effect on SNU-475 and SNU-423 cells with an IC50 value of 6.98 ± 0.11 μg/ml and 7.76 ± 0.45 μg/ml respectively, after 24 h of treatment. Significant dipping in the mitochondrial membrane potential and elevation in the released of cytochrome c from the mitochondria indicated the induction of the intrinsic apoptosis pathway by PCC. Activation of this pathway was further evidenced by significant activation of caspase 3/7 and 9. PCC was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-ƙB translocation to the nucleus. Cell cycle arrest in the G1 phase was confirmed by flow cytometry and up-regulation of glutathione reductase expression was quantified by qPCR. This study suggests that PCC is a simultaneous inducer of intrinsic and extrinsic pathways of apoptosis in liver cancer cell lines.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescence study of the effect of PCC.(A) After treatment of the cells with PCC (6.25 μg/ml), Hoechst 33342, cytochrome c, membrane permeability and mitochondrial membrane potential dyes were applied (B) Representative bar charts indicate a dose-dependent reduction of MMP, elevated cell permeability and cytochrome c release in treated cells. All data are expressed as the means ± standard error of triplicate measurements. *P < 0.05 compared with the no-treatment group.
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f3: Immunofluorescence study of the effect of PCC.(A) After treatment of the cells with PCC (6.25 μg/ml), Hoechst 33342, cytochrome c, membrane permeability and mitochondrial membrane potential dyes were applied (B) Representative bar charts indicate a dose-dependent reduction of MMP, elevated cell permeability and cytochrome c release in treated cells. All data are expressed as the means ± standard error of triplicate measurements. *P < 0.05 compared with the no-treatment group.

Mentions: Because of the cytotoxic effect of PCC on SNU-475 and SNU-423 cells, permeability of the membrane was higher than in the control suggesting sustained apoptotic activity in these cells (Fig. 3A,B). Loss of mitochondrial membrane potential (MMP) was evidenced as a conceivable mechanism for cell death using MMP dye. The cytoplasm of control cells was stained more intensely than the cells treated with PCC (Fig. 3A). Both SNU-475 and SNU-423 cells treated with PCC for 24 h exhibited a dose-dependent reduction of MMP fluorescence intensity, as a result of collapsed MMP (Fig. 3A,B). The fluorescence intensity in the cytosol of SNU-475 and SNU-423 cells treated with PCC was less than control cells suggesting the release of mitochondrial cytochrome c (Fig. 3B).


Novel piperazine core compound induces death in human liver cancer cells: possible pharmacological properties.

Samie N, Muniandy S, Kanthimathi MS, Haerian BS, Azudin RE - Sci Rep (2016)

Immunofluorescence study of the effect of PCC.(A) After treatment of the cells with PCC (6.25 μg/ml), Hoechst 33342, cytochrome c, membrane permeability and mitochondrial membrane potential dyes were applied (B) Representative bar charts indicate a dose-dependent reduction of MMP, elevated cell permeability and cytochrome c release in treated cells. All data are expressed as the means ± standard error of triplicate measurements. *P < 0.05 compared with the no-treatment group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829832&req=5

f3: Immunofluorescence study of the effect of PCC.(A) After treatment of the cells with PCC (6.25 μg/ml), Hoechst 33342, cytochrome c, membrane permeability and mitochondrial membrane potential dyes were applied (B) Representative bar charts indicate a dose-dependent reduction of MMP, elevated cell permeability and cytochrome c release in treated cells. All data are expressed as the means ± standard error of triplicate measurements. *P < 0.05 compared with the no-treatment group.
Mentions: Because of the cytotoxic effect of PCC on SNU-475 and SNU-423 cells, permeability of the membrane was higher than in the control suggesting sustained apoptotic activity in these cells (Fig. 3A,B). Loss of mitochondrial membrane potential (MMP) was evidenced as a conceivable mechanism for cell death using MMP dye. The cytoplasm of control cells was stained more intensely than the cells treated with PCC (Fig. 3A). Both SNU-475 and SNU-423 cells treated with PCC for 24 h exhibited a dose-dependent reduction of MMP fluorescence intensity, as a result of collapsed MMP (Fig. 3A,B). The fluorescence intensity in the cytosol of SNU-475 and SNU-423 cells treated with PCC was less than control cells suggesting the release of mitochondrial cytochrome c (Fig. 3B).

Bottom Line: PCC displayed a strong suppressive effect on SNU-475 and SNU-423 cells with an IC50 value of 6.98 ± 0.11 μg/ml and 7.76 ± 0.45 μg/ml respectively, after 24 h of treatment.Activation of this pathway was further evidenced by significant activation of caspase 3/7 and 9.Cell cycle arrest in the G1 phase was confirmed by flow cytometry and up-regulation of glutathione reductase expression was quantified by qPCR.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
The current study evaluates the cytotoxic mechanism of a novel piperazine derivate designated as PCC against human liver cancer cells. In this context, human liver cancer cell lines, SNU-475 and 243, human monocyte/macrophage cell line, CRL-9855, and human B lymphocyte cell line, CCL-156, were used to determine the IC50 of PCC using the standard MTT assay. PCC displayed a strong suppressive effect on SNU-475 and SNU-423 cells with an IC50 value of 6.98 ± 0.11 μg/ml and 7.76 ± 0.45 μg/ml respectively, after 24 h of treatment. Significant dipping in the mitochondrial membrane potential and elevation in the released of cytochrome c from the mitochondria indicated the induction of the intrinsic apoptosis pathway by PCC. Activation of this pathway was further evidenced by significant activation of caspase 3/7 and 9. PCC was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-ƙB translocation to the nucleus. Cell cycle arrest in the G1 phase was confirmed by flow cytometry and up-regulation of glutathione reductase expression was quantified by qPCR. This study suggests that PCC is a simultaneous inducer of intrinsic and extrinsic pathways of apoptosis in liver cancer cell lines.

No MeSH data available.


Related in: MedlinePlus