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Activation of endoplasmic reticulum stress is involved in the activity of icariin against human lung adenocarcinoma cells

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Abstract

In this study, we investigated the anticancer activity of icariin (ICA) against human lung adenocarcinoma cells in vitro and in vivo and explored the role of endoplasmic reticulum (ER) stress (ERS) signaling in this process. ICA treatment resulted in a dose- and time-dependent decrease in the viability of human lung adenocarcinoma A549 cells. Additionally, ICA exhibited potent anticancer activity, as evidenced by reductions in A549 cell adhesion, migration and intracellular glutathione (GSH) levels and increases in the apoptotic index, Caspase 3 activity, and reactive oxygen species. Furthermore, ICA treatment increased the expression of ERS-related molecules (p-PERK, ATF6, GRP78, p-eIF2α, and CHOP), up-regulated the apoptosis-related protein PUMA and down-regulated the anti-apoptosis-related protein Bcl2. The down-regulation of ERS signaling using PERK siRNA desensitized lung adenocarcinoma cells to ICA treatment, whereas the up-regulation of ERS signaling using thapsigargin (THA) sensitized lung adenocarcinoma cells to ICA treatment. Additionally, ICA inhibited the growth of human lung adenocarcinoma A549 cell xenografts by increasing the expression of ERS-related molecules (p-PERK and CHOP), up-regulating PUMA, and down-regulating Bcl2. These data indicate that ICA is a potential inhibitor of lung adenocarcinoma cell growth by targeting ERS signaling and suggest that the activation of ERS signaling may represent a novel therapeutic intervention for lung adenocarcinoma.

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (81000938) and the grants from the Excellent Doctoral Support Project of the Fourth Military Medical University (2013D01).

Conflict of interest

The authors declare that they have no conflicts of interest in the studies described.

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    Authors and Affiliations

    1. Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, 1 Xinsi Road, Xi’an, 710038, China

      Shouyin Di, Chongxi Fan, Yifang Zhu, Weimiao Li, Yongan Zhou, Xiaofei Li & Xiaolong Yan

    2. Department of Biomedical Engineering, The Fourth Military Medical University, 169 Changle West Road, Xi’an, 710032, China

      Yang Yang, Guiling Wu, Bodong Wang, Zhenlong Xin & Wei Hu

    3. Department of Aerospace Medicine, The Fourth Military Medical University, Xi’an, 710032, Shaanxi, China

      Shuai Jiang

    4. Department of Neurosurgery, Xijing Hospital, The Fourth Military Medical University, 127 Changle West Road, Xi’an, 710032, China

      Miaomiao Liang

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    1. Shouyin Di
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    Correspondence to Xiaofei Li or Xiaolong Yan.

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    Shouyin Di, Chongxi Fan and Yang Yang have contributed equally to this work.

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    10495_2015_1142_MOESM1_ESM.tif

    Supplementary Fig. 1 Effect of ICA treatment on the viability and morphology of human normal bronchial epithelial cells and lung adenocarcinoma cells. A. 16HBE cells were treated with different concentrations of ICA (25, 50, and 100 µM) and assessed at different time points (12, 24, and 36 h). Viability is expressed as OD values. B. A549 cells were treated with different concentrations of ICA (10, 15, and 20 µM) and assessed at different time points (12, 24, and 36 h). Viability is expressed as OD values. C. A549 cell morphology was observed under an inverted phase contrast microscope after the cells were treated for 24 h, and images were obtained. All of the results are expressed as the mean ± SD, n = 6. Supplementary material 1 (TIFF 2205 kb)

    10495_2015_1142_MOESM2_ESM.tif

    Supplementary Fig. 2 Effect of a lower concentration ICA on ERS signaling proteins in human lung adenocarcinoma cells (24 h). Representative Western blotting results of p-PERK, ATF6, GRP78, CHOP, and p-eIF2α are shown. Membranes were reprobed for β-actin expression to show that similar amounts of protein were loaded in each lane. The results are expressed as the mean ± SD, n = 6. aaP < 0.01 vs. the control group, bbP < 0.01 vs. the 10 μM ICA-treated group, ccP < 0.01 vs. the 15 μM ICA-treated group. Supplementary material 2 (TIFF 1080 kb)

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    Di, S., Fan, C., Yang, Y. et al. Activation of endoplasmic reticulum stress is involved in the activity of icariin against human lung adenocarcinoma cells. Apoptosis 20, 1229–1241 (2015). https://doi.org/10.1007/s10495-015-1142-0

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