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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 expression of cleaved caspase-8, caspase-9, caspase-3, and PARP protein in MA-10 cells.Notes: The cells were treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12 and 24 hours. After treatment, the cleaved caspase-8 (43/18 kDa) (A), caspase-9 (37 kDa) (B), caspase-3 (19 kDa) (C), and PARP (85–90 kDa) (D) proteins were detected by Western blot. The immunoblot represents the observations from one single experiment repeated at least three times. The integrated optical density of the cleaved protein was analyzed after normalization with β-actin (43 kDa) in each lane. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates that the means are significantly different compared with control (P<0.05).Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; T, paclitaxel; PARP, poly ADP-ribose polymerase.
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f3-ott-8-2345: Effects of cordycepin + paclitaxel and/or cisplatin on expression of cleaved caspase-8, caspase-9, caspase-3, and PARP protein in MA-10 cells.Notes: The cells were treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12 and 24 hours. After treatment, the cleaved caspase-8 (43/18 kDa) (A), caspase-9 (37 kDa) (B), caspase-3 (19 kDa) (C), and PARP (85–90 kDa) (D) proteins were detected by Western blot. The immunoblot represents the observations from one single experiment repeated at least three times. The integrated optical density of the cleaved protein was analyzed after normalization with β-actin (43 kDa) in each lane. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates that the means are significantly different compared with control (P<0.05).Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; T, paclitaxel; PARP, poly ADP-ribose polymerase.

Mentions: We then investigated whether cordycepin + paclitaxel and or cisplatin could activate the caspase cascade to induce apoptosis in MA-10 cells. Treatment with cordycepin alone for 12 or 24 hours significantly increased the expression of cleaved caspase-8 (P<0.05, Figure 3A). The combinations of cisplatin + cordycepin, cisplatin + paclitaxel, cordycepin + paclitaxel, and cisplatin + cordycepin + paclitaxel for 12 and 24 hours also induced significant caspase-8 cleavage (P<0.05, Figure 3A). Treatment with cordycepin (100 μM), paclitaxel (50 nM), cisplatin (100 μM), cisplatin + paclitaxel, cordycepin + paclitaxel, cisplatin + cordycepin, and cordycepin + cisplatin + paclitaxel for 12 and 24 hours all induced significant caspase-9 cleavage compared to the control (P<0.05). Indeed, treatment with cisplatin + cordycepin for 12 hours had a synergistic effect on cleavage of caspase-9. Cordycepin (100 μM), paclitaxel (50 nM), cisplatin (100 μM), cisplatin + paclitaxel, cordycepin + paclitaxel, cisplatin + cordycepin, and cordycepin + cisplatin + paclitaxel, but not cisplatin alone, induced significant caspase-3 cleavage compared with control after 24 hours of treatment (P<0.05, Figure 3C). Significant cleavage of PARP was observed after treatment with cordycepin for 12 and 24 hours, paclitaxel for 12 and 24 hours, cisplatin for 24 hours, cisplatin + cordycepin for 12 hours, cordycepin + paclitaxel for 12 and 24 hours, and cordycepin + cisplatin + paclitaxel for 12 and 24 hours (P<0.05, Figure 3D). These results indicate that cordycepin combined with paclitaxel and/or cisplatin can induce the intrinsic and extrinsic apoptotic pathways by activating cleavage of caspase-8, caspase-9, caspase-3, and PARP proteins.


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 expression of cleaved caspase-8, caspase-9, caspase-3, and PARP protein in MA-10 cells.Notes: The cells were treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12 and 24 hours. After treatment, the cleaved caspase-8 (43/18 kDa) (A), caspase-9 (37 kDa) (B), caspase-3 (19 kDa) (C), and PARP (85–90 kDa) (D) proteins were detected by Western blot. The immunoblot represents the observations from one single experiment repeated at least three times. The integrated optical density of the cleaved protein was analyzed after normalization with β-actin (43 kDa) in each lane. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates that the means are significantly different compared with control (P<0.05).Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; T, paclitaxel; PARP, poly ADP-ribose polymerase.
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f3-ott-8-2345: Effects of cordycepin + paclitaxel and/or cisplatin on expression of cleaved caspase-8, caspase-9, caspase-3, and PARP protein in MA-10 cells.Notes: The cells were treated without or with 50 nM paclitaxel and/or 100 μM cisplatin +100 μM cordycepin for 12 and 24 hours. After treatment, the cleaved caspase-8 (43/18 kDa) (A), caspase-9 (37 kDa) (B), caspase-3 (19 kDa) (C), and PARP (85–90 kDa) (D) proteins were detected by Western blot. The immunoblot represents the observations from one single experiment repeated at least three times. The integrated optical density of the cleaved protein was analyzed after normalization with β-actin (43 kDa) in each lane. Each datum point represents the mean ± standard error of the mean of three separate experiments. *Indicates that the means are significantly different compared with control (P<0.05).Abbreviations: Cor, cordycepin; Cis, cisplatin; DMSO, dimethyl sulfoxide; T, paclitaxel; PARP, poly ADP-ribose polymerase.
Mentions: We then investigated whether cordycepin + paclitaxel and or cisplatin could activate the caspase cascade to induce apoptosis in MA-10 cells. Treatment with cordycepin alone for 12 or 24 hours significantly increased the expression of cleaved caspase-8 (P<0.05, Figure 3A). The combinations of cisplatin + cordycepin, cisplatin + paclitaxel, cordycepin + paclitaxel, and cisplatin + cordycepin + paclitaxel for 12 and 24 hours also induced significant caspase-8 cleavage (P<0.05, Figure 3A). Treatment with cordycepin (100 μM), paclitaxel (50 nM), cisplatin (100 μM), cisplatin + paclitaxel, cordycepin + paclitaxel, cisplatin + cordycepin, and cordycepin + cisplatin + paclitaxel for 12 and 24 hours all induced significant caspase-9 cleavage compared to the control (P<0.05). Indeed, treatment with cisplatin + cordycepin for 12 hours had a synergistic effect on cleavage of caspase-9. Cordycepin (100 μM), paclitaxel (50 nM), cisplatin (100 μM), cisplatin + paclitaxel, cordycepin + paclitaxel, cisplatin + cordycepin, and cordycepin + cisplatin + paclitaxel, but not cisplatin alone, induced significant caspase-3 cleavage compared with control after 24 hours of treatment (P<0.05, Figure 3C). Significant cleavage of PARP was observed after treatment with cordycepin for 12 and 24 hours, paclitaxel for 12 and 24 hours, cisplatin for 24 hours, cisplatin + cordycepin for 12 hours, cordycepin + paclitaxel for 12 and 24 hours, and cordycepin + cisplatin + paclitaxel for 12 and 24 hours (P<0.05, Figure 3D). These results indicate that cordycepin combined with paclitaxel and/or cisplatin can induce the intrinsic and extrinsic apoptotic pathways by activating cleavage of caspase-8, caspase-9, caspase-3, and PARP proteins.

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