Resveratrol induces autophagic apoptosis via the lysosomal cathepsin D pathway in human drug‑resistant K562/ADM leukemia cells

Zhang,Zhewen; Liu,Zhuan; Chen,Jing; Yi,Juan; Cheng,Juan; Dun,Wangqing; Wei,Hulai

doi:10.3892/etm.2018.5742

Resveratrol induces autophagic apoptosis via the lysosomal cathepsin D pathway in human drug‑resistant K562/ADM leukemia cells

Authors:
- Zhewen Zhang
- Zhuan Liu
- Jing Chen
- Juan Yi
- Juan Cheng
- Wangqing Dun
- Hulai Wei
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Affiliations: Institute of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, Gansu 730000, P.R. China, Department of Hematology, The First Affiliated Hospital, Lanzhou University, Lanzhou, Gansu 730000, P.R. China
Published online on: January 12, 2018 https://doi.org/10.3892/etm.2018.5742
Pages: 3012-3019

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Abstract

The aim of the present study was to investigate the crosstalk between resveratrol (Res)‑induced autophagy and apoptosis, and the molecular pathway by which autophagy leads to apoptotic death in drug‑resistant K562/ADM leukemia cells. The viability of K562/ADM cells was determined using the MTT assay. The formation of autophagic vacuoles was detected using transmission electron microscopy and monodansylcadaverine (MDC) staining. Cell apoptosis was evaluated using flow cytometry. The expression of apoptosis‑ or autophagy‑associated proteins was measured using western blotting. The results indicated that treatment with Res inhibited cell viability in a concentration‑dependent manner. Furthermore, the numbers of MDC‑positive fluorescent points, autophagic vacuoles and autolysosome‑engulfed cytoplasmic materials were markedly increased in Res‑treated K562/ADM cells compared with untreated cells, as determined using fluorescence microscopy and transmission electron microscopy. Res‑induced apoptosis was associated with increased cleaved caspase‑3 and B‑cell lymphoma 2 associated X protein, and decreased B‑cell lymphoma 2 (Bcl‑2) protein expression levels when compared with the control (P<0.05). However, the proportion of apoptotic cells decreased from 69.6 to 41.0% (40 µmol/l Res) and from 77.3 to 58.8% (80 µmol/l Res) following pre‑treatment with the autophagy inhibitor 3‑methyladenine (P<0.01). The protein expression levels of microtubule‑associated protein 1A/1B‑light chain 3 and beclin 1, two markers of autophagy, were upregulated in Res‑treated cells compared with the control (P<0.05). In addition, lysosomal cathepsin D (Cath D) release increased during Res‑induced autophagy and apoptosis (P<0.05). The present results demonstrated that Res‑induced apoptosis of K562/ADM cells was autophagy‑dependent and the released Cath D may trigger caspase‑dependent cell death through the Bcl‑2 family of proteins. Furthermore, the present data indicate that to enhancement of the autophagy‑cathepsin‑apoptosis pathway may be an effective approach for overcoming anticancer drug resistance.

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