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LYG-202 exerts antitumor effect on PI3K/Akt signaling pathway in human breast cancer cells

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Abstract

In this study, we aimed to investigate the antitumor effect of LYG-202, a newly synthesized piperazine-substituted derivative of flavonoid on human breast cancer cells and illustrate the potential mechanisms. LYG-202 induced apoptosis in MCF-7, MDA-MB-231 and MDA-MB-435 cells. LYG-202 triggered the activation of mitochondrial apoptotic pathway through multiple steps: increasing Bax/Bcl-2 ratio, decreasing mitochondrial membrane potential (ΔΨ m ), activating caspase-9 and caspase-3, inducing cleavage of poly(ADP-ribose) polymerase, cytochrome c release and apoptosis-inducing factor translocation. Furthermore, LYG-202 inhibited cell cycle progression at the G1/S transition via targeting Cyclin D, CDK4 and p21Waf1/Cip1. Additionally, LYG-202 increased the generation of intracellular ROS. N-Acetyl cysteine, an antioxidant, reversed LYG-202-induced apoptosis suggesting that LYG-202 induces apoptosis by accelerating ROS generation. Further, we found that LYG-202 deactivated the PI3K/Akt pathway, activated Bad phosphorylation, increased Cyclin D and Bcl-xL expression, and inhibited NF-κB nuclear translocation. Activation of PI3K/Akt pathway by IGF-1 attenuated LYG-202-induced apoptosis and cell cycle arrest. Our in vivo study showed that LYG-202 exhibited a potential antitumor effect in nude mice inoculated with MCF-7 tumor through similar mechanisms identified in cultured cells. In summary, our results demonstrated that LYG-202 induced apoptosis and cell cycle arrest via targeting PI3K/Akt pathway, indicating that LYG-202 is a potential anticancer agent for breast cancer.

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Abbreviations

ROS:

Reactive oxygen species

DAPI:

4′,6-Diamidino-2-phenylindole

DMSO:

Dimethylsulfoxide

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PBS:

Phosphate buffered saline

PI:

Propidium iodide

PARP:

Poly(ADP-ribose) polymerase

AIF:

Apoptosis-inducing factor

CDK:

Cyclin-dependent kinase

NAC:

N-Acetyl cysteine

IGF-1:

Insulin-like growth factor-1

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Acknowledgments

This work was supported by the Fundamental Research Funds for the Central Universities (No. JKY2011051), the National Natural Science Foundation of China (No. 21072232), the Project Program of State Key Laboratory of Natural Medicines, China Pharmaceutical University (No. JKGZ201101), the National Science & Technology Major Project (No. 2012ZX09304-001) and Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT-IRT1193).

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The authors declare that they have no competing interests.

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    Authors and Affiliations

    1. State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, Jiangsu Key Laboratory of Drug Design and Optimization, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, People’s Republic of China

      Yue Zhao, Xiaoping Wang, Yang Sun, Yuxin Zhou, Yuehan Yin, Youxiang Ding, Zhiyu Li, Qinglong Guo & Na Lu

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    1. Yue Zhao
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    Correspondence to Qinglong Guo or Na Lu.

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    Yue Zhao and Xiaoping Wang contributed equally to this work.

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    Zhao, Y., Wang, X., Sun, Y. et al. LYG-202 exerts antitumor effect on PI3K/Akt signaling pathway in human breast cancer cells. Apoptosis 20, 1253–1269 (2015). https://doi.org/10.1007/s10495-015-1145-x

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