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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

JNK inhibitor (SP600125) reversed phosphorylation of JNK induced by cordycepin + paclitaxel and/or cisplatin in MA-10 cells.Notes: The cells were pretreated with 1, 10, 50, and 100 μM SP600125 for 30 minutes, and then treated without or with 100 μM cordycepin and 1, 10, 50, and 100 μM SP600125 for 12 hours (A), or without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin and 10 μM SP600125 for 12 hours (B). Phosphorylated JNK (54/46 kDa) and t-JNK (54/46 kDa) were then detected by Western blots. The integrated optical density of p-JNK protein expression after normalization with t-JNK is demonstrated. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates a statistically significant difference compared with the control (P<0.05). #Indicates that the means are significantly (P<0.05) different compared with treatment.Abbreviations: p-, phosphorylated; t-, total; Cor, cordycepin; Cis, cisplatin; SP, SP600125; DMSO, dimethyl sulfoxide; JNK, c-Jun NH2-terminal kinase.
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f5-ott-8-2345: JNK inhibitor (SP600125) reversed phosphorylation of JNK induced by cordycepin + paclitaxel and/or cisplatin in MA-10 cells.Notes: The cells were pretreated with 1, 10, 50, and 100 μM SP600125 for 30 minutes, and then treated without or with 100 μM cordycepin and 1, 10, 50, and 100 μM SP600125 for 12 hours (A), or without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin and 10 μM SP600125 for 12 hours (B). Phosphorylated JNK (54/46 kDa) and t-JNK (54/46 kDa) were then detected by Western blots. The integrated optical density of p-JNK protein expression after normalization with t-JNK is demonstrated. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates a statistically significant difference compared with the control (P<0.05). #Indicates that the means are significantly (P<0.05) different compared with treatment.Abbreviations: p-, phosphorylated; t-, total; Cor, cordycepin; Cis, cisplatin; SP, SP600125; DMSO, dimethyl sulfoxide; JNK, c-Jun NH2-terminal kinase.

Mentions: To reconfirm whether phosphorylation of JNK, ERK, and p38 proteins could be induced by cordycepin + paclitaxel and/or cisplatin in MA-10 cells, inhibitors of JNK, ERK, and p38 (SP600125, PD98059, and SB203580, respectively) were used to abolish expression of phosphorylated JNK, ERK, and p38. SP600125 at 10, 50, and 100 μM significantly inhibited phosphorylation of JNK induced by cordycepin 100 μM (P<0.05, Figure 5A). In the combination experiments, SP600125 10 μM significantly reduced the expression of phosphorylated JNK after treatment with cisplatin + cordycepin, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel for 12 hours (P<0.05, Figure 5B). PD98059 at 5, 10, 25, and 50 μM significantly inhibited phosphorylation of ERK induced by cordycepin 100 μM (P<0.05, Figure 6A). In the combination experiments, PD98059 5 μM significantly reduced the expression of phosphorylated ERK in the cisplatin + cordycepin, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel groups treated for 12 hours (P<0.05, Figure 6B). SB203580 at 1, 5, and 10 μM significantly inhibited the phosphorylation of p38 induced by 100 μM cordycepin (P<0.05, Figure 7A). In the combination experiments, SB203580 10 μM significantly reduced the expression of phosphorylated p38 in the cisplatin + cordycepin, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel groups (P<0.05, Figure 7A and B). These results again showed that cordycepin + paclitaxel and/or cisplatin could stimulate activation of the JNK, ERK1/2, and/or p38 MAPK pathways to induce apoptosis of MA-10 cells.


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)

JNK inhibitor (SP600125) reversed phosphorylation of JNK induced by cordycepin + paclitaxel and/or cisplatin in MA-10 cells.Notes: The cells were pretreated with 1, 10, 50, and 100 μM SP600125 for 30 minutes, and then treated without or with 100 μM cordycepin and 1, 10, 50, and 100 μM SP600125 for 12 hours (A), or without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin and 10 μM SP600125 for 12 hours (B). Phosphorylated JNK (54/46 kDa) and t-JNK (54/46 kDa) were then detected by Western blots. The integrated optical density of p-JNK protein expression after normalization with t-JNK is demonstrated. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates a statistically significant difference compared with the control (P<0.05). #Indicates that the means are significantly (P<0.05) different compared with treatment.Abbreviations: p-, phosphorylated; t-, total; Cor, cordycepin; Cis, cisplatin; SP, SP600125; DMSO, dimethyl sulfoxide; JNK, c-Jun NH2-terminal kinase.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4562734&req=5

f5-ott-8-2345: JNK inhibitor (SP600125) reversed phosphorylation of JNK induced by cordycepin + paclitaxel and/or cisplatin in MA-10 cells.Notes: The cells were pretreated with 1, 10, 50, and 100 μM SP600125 for 30 minutes, and then treated without or with 100 μM cordycepin and 1, 10, 50, and 100 μM SP600125 for 12 hours (A), or without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin and 10 μM SP600125 for 12 hours (B). Phosphorylated JNK (54/46 kDa) and t-JNK (54/46 kDa) were then detected by Western blots. The integrated optical density of p-JNK protein expression after normalization with t-JNK is demonstrated. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates a statistically significant difference compared with the control (P<0.05). #Indicates that the means are significantly (P<0.05) different compared with treatment.Abbreviations: p-, phosphorylated; t-, total; Cor, cordycepin; Cis, cisplatin; SP, SP600125; DMSO, dimethyl sulfoxide; JNK, c-Jun NH2-terminal kinase.
Mentions: To reconfirm whether phosphorylation of JNK, ERK, and p38 proteins could be induced by cordycepin + paclitaxel and/or cisplatin in MA-10 cells, inhibitors of JNK, ERK, and p38 (SP600125, PD98059, and SB203580, respectively) were used to abolish expression of phosphorylated JNK, ERK, and p38. SP600125 at 10, 50, and 100 μM significantly inhibited phosphorylation of JNK induced by cordycepin 100 μM (P<0.05, Figure 5A). In the combination experiments, SP600125 10 μM significantly reduced the expression of phosphorylated JNK after treatment with cisplatin + cordycepin, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel for 12 hours (P<0.05, Figure 5B). PD98059 at 5, 10, 25, and 50 μM significantly inhibited phosphorylation of ERK induced by cordycepin 100 μM (P<0.05, Figure 6A). In the combination experiments, PD98059 5 μM significantly reduced the expression of phosphorylated ERK in the cisplatin + cordycepin, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel groups treated for 12 hours (P<0.05, Figure 6B). SB203580 at 1, 5, and 10 μM significantly inhibited the phosphorylation of p38 induced by 100 μM cordycepin (P<0.05, Figure 7A). In the combination experiments, SB203580 10 μM significantly reduced the expression of phosphorylated p38 in the cisplatin + cordycepin, cordycepin + paclitaxel, and cordycepin + cisplatin + paclitaxel groups (P<0.05, Figure 7A and B). These results again showed that cordycepin + paclitaxel and/or cisplatin could stimulate activation of the JNK, ERK1/2, and/or p38 MAPK pathways to induce apoptosis of MA-10 cells.

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