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Systems biology network-based discovery of a small molecule activator BL-AD008 targeting AMPK/ZIPK and inducing apoptosis in cervical cancer.

Fu L, Zhang S, Zhang L, Tong X, Zhang J, Zhang Y, Ouyang L, Liu B, Huang J - Oncotarget (2015)

Bottom Line: Subsequently, we screened a series of candidate compounds targeting AMPK/ZIPK, synthesized some compounds and eventually discovered a novel dual-target activator (BL-AD008).Additionally, we found that BL-AD008-induced apoptosis was affected by the combination of AMPK and ZIPK.Then, we found that BL-AD008 bear its anti-tumor activities and induced apoptosis by targeting AMPK/ZIPK in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Biotherapy, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.

ABSTRACT
The aim of this study was to discover a small molecule activator BL-AD008 targeting AMPK/ZIPK and inducing apoptosis in cervical cancer. In this study, we systematically constructed the global protein-protein interaction (PPI) network and predicted apoptosis-related protein connections by the Naïve Bayesian model. Then, we identified some classical apoptotic PPIs and other previously unrecognized PPIs between apoptotic kinases, such as AMPK and ZIPK. Subsequently, we screened a series of candidate compounds targeting AMPK/ZIPK, synthesized some compounds and eventually discovered a novel dual-target activator (BL-AD008). Moreover, we found BL-AD008 bear remarkable anti-proliferative activities toward cervical cancer cells and could induce apoptosis by death-receptor and mitochondrial pathways. Additionally, we found that BL-AD008-induced apoptosis was affected by the combination of AMPK and ZIPK. Then, we found that BL-AD008 bear its anti-tumor activities and induced apoptosis by targeting AMPK/ZIPK in vivo. In conclusion, these results demonstrate the ability of systems biology network to identify some key apoptotic kinase targets AMPK and ZIPK; thus providing a dual-target small molecule activator (BL-AD008) as a potential new apoptosis-modulating drug in future cervical cancer therapy.

No MeSH data available.


Related in: MedlinePlus

BL-AD008 induces apoptosis in vivo(A) Immunohistochemistry of cleaved caspase-3 and -8, Bcl-2, Bax, ZIPK, p-AMPKα. IHC staining of the mouse orthotopic tumor tissues. IHC was used to determine the expression levels of apoptosis markers, which are cleaved caspase-3 and -8, Bcl-2 and Bax. And the ZIPK, p-AMPKα levels increasing (×200 magnification). Tumor tissues excised from the median dose group treated mice; *, P <0.05; **, P <0.01; ***, P<0.001. (b) Western blot analysis of AMPK, ZIPK, ERK1, cleaved caspase-3, -8 and -9. Tumor tissues excised from the HeLa xenograft mice were lysed.
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Figure 9: BL-AD008 induces apoptosis in vivo(A) Immunohistochemistry of cleaved caspase-3 and -8, Bcl-2, Bax, ZIPK, p-AMPKα. IHC staining of the mouse orthotopic tumor tissues. IHC was used to determine the expression levels of apoptosis markers, which are cleaved caspase-3 and -8, Bcl-2 and Bax. And the ZIPK, p-AMPKα levels increasing (×200 magnification). Tumor tissues excised from the median dose group treated mice; *, P <0.05; **, P <0.01; ***, P<0.001. (b) Western blot analysis of AMPK, ZIPK, ERK1, cleaved caspase-3, -8 and -9. Tumor tissues excised from the HeLa xenograft mice were lysed.

Mentions: For better understanding of the mechanism of the therapeutic efficacy of BL-AD008 in our in vivo model, we examined the caspase-3, caspase-8, Bcl-2, Bax, p-AMPKα and ZIPK expressions in tumor samples immunoreactivity. Active form of caspase-3 and caspase-8 were observed in the tumor. And, Bcl-2 was inhibited by BL-AD008 while Bax expression was increased (Figure 9A). They were determined in tumor xenografts as parameters for the apoptosis levels. Western blotting assay further demonstrated that all their expression in BL-AD008-treated tumor samples were consistent with immunohistochemical results and caspase cascade activating. And we also tested the p-AMPKα and ZIPK for their regulation by BL-AD008. In addition, DNA repairing protein PARP-1 were sheared significantly (Figure 9B). The increase of the expression levels of p-AMPKα and ZIPK confirmed the efficacy of BL-AD008 in tumor tissues. Moreover, immunohistochemical and western blotting results suggesting the apoptosis induced by BL-AD008 in tumor tissues.


Systems biology network-based discovery of a small molecule activator BL-AD008 targeting AMPK/ZIPK and inducing apoptosis in cervical cancer.

Fu L, Zhang S, Zhang L, Tong X, Zhang J, Zhang Y, Ouyang L, Liu B, Huang J - Oncotarget (2015)

BL-AD008 induces apoptosis in vivo(A) Immunohistochemistry of cleaved caspase-3 and -8, Bcl-2, Bax, ZIPK, p-AMPKα. IHC staining of the mouse orthotopic tumor tissues. IHC was used to determine the expression levels of apoptosis markers, which are cleaved caspase-3 and -8, Bcl-2 and Bax. And the ZIPK, p-AMPKα levels increasing (×200 magnification). Tumor tissues excised from the median dose group treated mice; *, P <0.05; **, P <0.01; ***, P<0.001. (b) Western blot analysis of AMPK, ZIPK, ERK1, cleaved caspase-3, -8 and -9. Tumor tissues excised from the HeLa xenograft mice were lysed.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4480736&req=5

Figure 9: BL-AD008 induces apoptosis in vivo(A) Immunohistochemistry of cleaved caspase-3 and -8, Bcl-2, Bax, ZIPK, p-AMPKα. IHC staining of the mouse orthotopic tumor tissues. IHC was used to determine the expression levels of apoptosis markers, which are cleaved caspase-3 and -8, Bcl-2 and Bax. And the ZIPK, p-AMPKα levels increasing (×200 magnification). Tumor tissues excised from the median dose group treated mice; *, P <0.05; **, P <0.01; ***, P<0.001. (b) Western blot analysis of AMPK, ZIPK, ERK1, cleaved caspase-3, -8 and -9. Tumor tissues excised from the HeLa xenograft mice were lysed.
Mentions: For better understanding of the mechanism of the therapeutic efficacy of BL-AD008 in our in vivo model, we examined the caspase-3, caspase-8, Bcl-2, Bax, p-AMPKα and ZIPK expressions in tumor samples immunoreactivity. Active form of caspase-3 and caspase-8 were observed in the tumor. And, Bcl-2 was inhibited by BL-AD008 while Bax expression was increased (Figure 9A). They were determined in tumor xenografts as parameters for the apoptosis levels. Western blotting assay further demonstrated that all their expression in BL-AD008-treated tumor samples were consistent with immunohistochemical results and caspase cascade activating. And we also tested the p-AMPKα and ZIPK for their regulation by BL-AD008. In addition, DNA repairing protein PARP-1 were sheared significantly (Figure 9B). The increase of the expression levels of p-AMPKα and ZIPK confirmed the efficacy of BL-AD008 in tumor tissues. Moreover, immunohistochemical and western blotting results suggesting the apoptosis induced by BL-AD008 in tumor tissues.

Bottom Line: Subsequently, we screened a series of candidate compounds targeting AMPK/ZIPK, synthesized some compounds and eventually discovered a novel dual-target activator (BL-AD008).Additionally, we found that BL-AD008-induced apoptosis was affected by the combination of AMPK and ZIPK.Then, we found that BL-AD008 bear its anti-tumor activities and induced apoptosis by targeting AMPK/ZIPK in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Biotherapy, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.

ABSTRACT
The aim of this study was to discover a small molecule activator BL-AD008 targeting AMPK/ZIPK and inducing apoptosis in cervical cancer. In this study, we systematically constructed the global protein-protein interaction (PPI) network and predicted apoptosis-related protein connections by the Naïve Bayesian model. Then, we identified some classical apoptotic PPIs and other previously unrecognized PPIs between apoptotic kinases, such as AMPK and ZIPK. Subsequently, we screened a series of candidate compounds targeting AMPK/ZIPK, synthesized some compounds and eventually discovered a novel dual-target activator (BL-AD008). Moreover, we found BL-AD008 bear remarkable anti-proliferative activities toward cervical cancer cells and could induce apoptosis by death-receptor and mitochondrial pathways. Additionally, we found that BL-AD008-induced apoptosis was affected by the combination of AMPK and ZIPK. Then, we found that BL-AD008 bear its anti-tumor activities and induced apoptosis by targeting AMPK/ZIPK in vivo. In conclusion, these results demonstrate the ability of systems biology network to identify some key apoptotic kinase targets AMPK and ZIPK; thus providing a dual-target small molecule activator (BL-AD008) as a potential new apoptosis-modulating drug in future cervical cancer therapy.

No MeSH data available.


Related in: MedlinePlus