Microcystin-LR induces mitochondria-mediated apoptosis in human bronchial epithelial cells

Li,Yang; Li,Jinhui; Huang,Hui; Yang,Mingfeng; Zhuang,Donggang; Cheng,Xuemin; Zhang,Huizhen; Fu,Xiaoli

doi:10.3892/etm.2016.3423

Microcystin-LR induces mitochondria-mediated apoptosis in human bronchial epithelial cells

Authors:
- Yang Li
- Jinhui Li
- Hui Huang
- Mingfeng Yang
- Donggang Zhuang
- Xuemin Cheng
- Huizhen Zhang
- Xiaoli Fu
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Affiliations: College of Public Health, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China, Henan Science and Technology Exchange Center with Foreign Countries, Zhengzhou University, Zhengzhou, Henan 450003, P.R. China
Published online on: June 3, 2016 https://doi.org/10.3892/etm.2016.3423
Pages: 633-640
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate the toxicity of microcystin-LR (MC-LR) and to explore the mechanism of MC-LR-induced apoptosis in human bronchial epithelial (HBE) cells. HBE cells were treated with MC‑LR (1, 10, 20, 30 and 40 µg/ml) alone or with MC‑LR (0, 2.5, 5 and 10 µg/ml) and Z-VAD-FMK (0, 10, 20, 40, 60, 80, 100, 120 and 140 µM), which is a caspase inhibitor, for 24 and 48 h. Cell viability was assessed via an MTT assay and the half maximal effective concentration of MC‑LR was determined. The optimal concentration of Z‑VAD‑FMK was established as 50 µm, which was then used in the subsequent experiments. MC‑LR significantly inhibited cell viability and induced apoptosis of HBE cells in a dose‑dependent manner, as detected by an Annexin V/propidium iodide assay. MC‑LR induced cell apoptosis, excess reactive oxygen species production and mitochondrial membrane potential collapse, upregulated Bax expression and downregulated B‑cell lymphoma‑2 expression in HBE cells. Moreover, western blot analysis demonstrated that MC‑LR increased the activity levels of caspase‑3 and caspase‑9 and induced cytochrome c release into the cytoplasm, suggesting that MC‑LR‑induced apoptosis is associated with the mitochondrial pathway. Furthermore, pretreatment with Z-VAD-FMK reduced MC‑LR‑induced apoptosis by blocking caspase activation in HBE cells. Therefore, the results of the present study suggested that MC‑LR is capable of significantly inhibiting the viability of HBE cells by inducing apoptosis in a mitochondria-dependent manner. The present study provides a foundation for further understanding the mechanism underlying the toxicity of MC-LR in the respiratory system.

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