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Apoptotic effect of cordycepin combined with cisplatin and/or paclitaxel on MA-10 mouse Leydig tumor cells.

Kang FC, Chen PJ, Pan BS, Lai MS, Chen YC, Huang BM - Onco Targets Ther (2015)

Bottom Line: Cell cycle analysis showed that combination treatment significantly increased subG1 phase cell numbers in MA-10 cells, indicating apoptosis.Moreover, cordycepin plus cisplatin and/or paclitaxel significantly induced cleavage of caspase-8, caspase-9, caspase-3, and poly ADP-ribose polymerase, and phosphorylation of c-Jun NH2-terminal kinase, extracellular signal-regulated kinase, p38, and p53 proteins in MA-10 cells.Cordycepin plus cisplatin and/or paclitaxel can have an additive effect on apoptosis in MA-10 cells, with activation of caspase, mitogen-activated protein kinase, and p53 signal pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesia, Chi Mei Medical Center, Chiali, Republic of China.

ABSTRACT

Background: Chemotherapy is not limited to a single treatment, and the evidence demonstrates that different drug combinations can have positive results in patients. In this study, we sought to determine whether cordycepin combined with cisplatin and/or paclitaxel would have an additive effective on inducing apoptosis in mouse Leydig tumor cells, and the mechanisms were also briefly examined.

Methods: The additive effects of cordycepin combined with cisplatin and/or paclitaxel on apoptosis in MA-10 cells were investigated by monitoring changes in morphological characteristics and examining cell viability, flow cytometry assays, and Western blot analyses.

Results: Combination of cordycepin plus cisplatin and/or paclitaxel for 12 and 24 hours induced apoptotic features in MA-10 cells. The MTT assay showed that the combination treatment reduced the viability of MA-10 cells in a dose-dependent manner, with additive effects. Cell cycle analysis showed that combination treatment significantly increased subG1 phase cell numbers in MA-10 cells, indicating apoptosis. Moreover, cordycepin plus cisplatin and/or paclitaxel significantly induced cleavage of caspase-8, caspase-9, caspase-3, and poly ADP-ribose polymerase, and phosphorylation of c-Jun NH2-terminal kinase, extracellular signal-regulated kinase, p38, and p53 proteins in MA-10 cells.

Conclusion: Cordycepin plus cisplatin and/or paclitaxel can have an additive effect on apoptosis in MA-10 cells, with activation of caspase, mitogen-activated protein kinase, and p53 signal pathways.

No MeSH data available.


Related in: MedlinePlus

Effects of cordycepin + paclitaxel and/or cisplatin on morphology and viability of MA-10 cells.Notes: Cells were cultured in 6 cm dishes until 70%–80% confluence, and then treated without or with paclitaxel and/or cisplatin + cordycepin for 24 hours. (A) Control, (B) 0.5% DMSO, (C) 100 μM cordycepin, (D) 50 nM paclitaxel, (E) 100 μM cisplatin, (F) 100 μM cisplatin +100 μM cordycepin, (G) 100 μM cisplatin +50 nM paclitaxel, (H) 50 nM paclitaxel +100 μM cordycepin, and (I) 100 μM cordycepin +100 μM cisplatin +50 nM paclitaxel. Morphological changes in cells were observed by light microscopy (magnification 200×, arrowheads indicate cells with membrane blebbing; insets show cells with membrane blebbing under larger magnification). The experiments were performed three times with similar results. In the MTT viability assay, cells were cultured in 96-well plates until 70%–80% confluence, and then treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12, 24, 36, or 48 hours. (J) The results are expressed as the percentage of cell growth relative to control groups as 100%. *Indicates that the means are significantly different when compared with control (P<0.05). The experiments were performed three times with similar results.Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
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f1-ott-8-2345: Effects of cordycepin + paclitaxel and/or cisplatin on morphology and viability of MA-10 cells.Notes: Cells were cultured in 6 cm dishes until 70%–80% confluence, and then treated without or with paclitaxel and/or cisplatin + cordycepin for 24 hours. (A) Control, (B) 0.5% DMSO, (C) 100 μM cordycepin, (D) 50 nM paclitaxel, (E) 100 μM cisplatin, (F) 100 μM cisplatin +100 μM cordycepin, (G) 100 μM cisplatin +50 nM paclitaxel, (H) 50 nM paclitaxel +100 μM cordycepin, and (I) 100 μM cordycepin +100 μM cisplatin +50 nM paclitaxel. Morphological changes in cells were observed by light microscopy (magnification 200×, arrowheads indicate cells with membrane blebbing; insets show cells with membrane blebbing under larger magnification). The experiments were performed three times with similar results. In the MTT viability assay, cells were cultured in 96-well plates until 70%–80% confluence, and then treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12, 24, 36, or 48 hours. (J) The results are expressed as the percentage of cell growth relative to control groups as 100%. *Indicates that the means are significantly different when compared with control (P<0.05). The experiments were performed three times with similar results.Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.

Mentions: MA-10 cells were treated with 100 μM cordycepin, 50 nM paclitaxel, or 100 μM cisplatin alone or in combination with cordycepin + paclitaxel and/or cisplatin without serum for 24 hours. The changes in morphology were then observed under a light microscope. Cells in the control and in DMSO demonstrated a normal polygonal shape, with firm attachment to the culture dishes, which is a normal cell growth phenomenon (Figure 1A and B). After 24 hours, cells treated with 100 μM cordycepin, 50 nM paclitaxel, or 100 μM cisplatin had a rounded-up appearance but still adhered to the matrix (Figure 1C–E). Treatment for 24 hours with combinations of cisplatin + cordycepin, cisplatin + paclitaxel, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel resulted in more loss of cell attachment to the matrix, more membrane blebbing, and many more floating cells (Figure 1F–I). The morphological study suggested that the combination treatments induced more cell death in MA-10 cells than treatment with cordycepin, paclitaxel, or cisplatin alone.


Apoptotic effect of cordycepin combined with cisplatin and/or paclitaxel on MA-10 mouse Leydig tumor cells.

Kang FC, Chen PJ, Pan BS, Lai MS, Chen YC, Huang BM - Onco Targets Ther (2015)

Effects of cordycepin + paclitaxel and/or cisplatin on morphology and viability of MA-10 cells.Notes: Cells were cultured in 6 cm dishes until 70%–80% confluence, and then treated without or with paclitaxel and/or cisplatin + cordycepin for 24 hours. (A) Control, (B) 0.5% DMSO, (C) 100 μM cordycepin, (D) 50 nM paclitaxel, (E) 100 μM cisplatin, (F) 100 μM cisplatin +100 μM cordycepin, (G) 100 μM cisplatin +50 nM paclitaxel, (H) 50 nM paclitaxel +100 μM cordycepin, and (I) 100 μM cordycepin +100 μM cisplatin +50 nM paclitaxel. Morphological changes in cells were observed by light microscopy (magnification 200×, arrowheads indicate cells with membrane blebbing; insets show cells with membrane blebbing under larger magnification). The experiments were performed three times with similar results. In the MTT viability assay, cells were cultured in 96-well plates until 70%–80% confluence, and then treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12, 24, 36, or 48 hours. (J) The results are expressed as the percentage of cell growth relative to control groups as 100%. *Indicates that the means are significantly different when compared with control (P<0.05). The experiments were performed three times with similar results.Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
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f1-ott-8-2345: Effects of cordycepin + paclitaxel and/or cisplatin on morphology and viability of MA-10 cells.Notes: Cells were cultured in 6 cm dishes until 70%–80% confluence, and then treated without or with paclitaxel and/or cisplatin + cordycepin for 24 hours. (A) Control, (B) 0.5% DMSO, (C) 100 μM cordycepin, (D) 50 nM paclitaxel, (E) 100 μM cisplatin, (F) 100 μM cisplatin +100 μM cordycepin, (G) 100 μM cisplatin +50 nM paclitaxel, (H) 50 nM paclitaxel +100 μM cordycepin, and (I) 100 μM cordycepin +100 μM cisplatin +50 nM paclitaxel. Morphological changes in cells were observed by light microscopy (magnification 200×, arrowheads indicate cells with membrane blebbing; insets show cells with membrane blebbing under larger magnification). The experiments were performed three times with similar results. In the MTT viability assay, cells were cultured in 96-well plates until 70%–80% confluence, and then treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12, 24, 36, or 48 hours. (J) The results are expressed as the percentage of cell growth relative to control groups as 100%. *Indicates that the means are significantly different when compared with control (P<0.05). The experiments were performed three times with similar results.Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
Mentions: MA-10 cells were treated with 100 μM cordycepin, 50 nM paclitaxel, or 100 μM cisplatin alone or in combination with cordycepin + paclitaxel and/or cisplatin without serum for 24 hours. The changes in morphology were then observed under a light microscope. Cells in the control and in DMSO demonstrated a normal polygonal shape, with firm attachment to the culture dishes, which is a normal cell growth phenomenon (Figure 1A and B). After 24 hours, cells treated with 100 μM cordycepin, 50 nM paclitaxel, or 100 μM cisplatin had a rounded-up appearance but still adhered to the matrix (Figure 1C–E). Treatment for 24 hours with combinations of cisplatin + cordycepin, cisplatin + paclitaxel, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel resulted in more loss of cell attachment to the matrix, more membrane blebbing, and many more floating cells (Figure 1F–I). The morphological study suggested that the combination treatments induced more cell death in MA-10 cells than treatment with cordycepin, paclitaxel, or cisplatin alone.

Bottom Line: Cell cycle analysis showed that combination treatment significantly increased subG1 phase cell numbers in MA-10 cells, indicating apoptosis.Moreover, cordycepin plus cisplatin and/or paclitaxel significantly induced cleavage of caspase-8, caspase-9, caspase-3, and poly ADP-ribose polymerase, and phosphorylation of c-Jun NH2-terminal kinase, extracellular signal-regulated kinase, p38, and p53 proteins in MA-10 cells.Cordycepin plus cisplatin and/or paclitaxel can have an additive effect on apoptosis in MA-10 cells, with activation of caspase, mitogen-activated protein kinase, and p53 signal pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesia, Chi Mei Medical Center, Chiali, Republic of China.

ABSTRACT

Background: Chemotherapy is not limited to a single treatment, and the evidence demonstrates that different drug combinations can have positive results in patients. In this study, we sought to determine whether cordycepin combined with cisplatin and/or paclitaxel would have an additive effective on inducing apoptosis in mouse Leydig tumor cells, and the mechanisms were also briefly examined.

Methods: The additive effects of cordycepin combined with cisplatin and/or paclitaxel on apoptosis in MA-10 cells were investigated by monitoring changes in morphological characteristics and examining cell viability, flow cytometry assays, and Western blot analyses.

Results: Combination of cordycepin plus cisplatin and/or paclitaxel for 12 and 24 hours induced apoptotic features in MA-10 cells. The MTT assay showed that the combination treatment reduced the viability of MA-10 cells in a dose-dependent manner, with additive effects. Cell cycle analysis showed that combination treatment significantly increased subG1 phase cell numbers in MA-10 cells, indicating apoptosis. Moreover, cordycepin plus cisplatin and/or paclitaxel significantly induced cleavage of caspase-8, caspase-9, caspase-3, and poly ADP-ribose polymerase, and phosphorylation of c-Jun NH2-terminal kinase, extracellular signal-regulated kinase, p38, and p53 proteins in MA-10 cells.

Conclusion: Cordycepin plus cisplatin and/or paclitaxel can have an additive effect on apoptosis in MA-10 cells, with activation of caspase, mitogen-activated protein kinase, and p53 signal pathways.

No MeSH data available.


Related in: MedlinePlus