Involvement of β‑catenin in matrine‑induced autophagy and apoptosis in WB‑F344 cells

Xie,Bu‑Shan; He,Xing‑Xing; Ai,Zheng‑Lin; Yao,Shu‑Kun

doi:10.3892/mmr.2014.2107

Involvement of β‑catenin in matrine‑induced autophagy and apoptosis in WB‑F344 cells

Authors:
- Bu‑Shan Xie
- Xing‑Xing He
- Zheng‑Lin Ai
- Shu‑Kun Yao
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Affiliations: Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, P.R. China, Graduate School, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, P.R. China, Department of Gastroenterology, China‑Japan Friendship Hospital, Beijing 100029, P.R. China
Published online on: April 2, 2014 https://doi.org/10.3892/mmr.2014.2107
Pages: 2547-2553

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Abstract

Matrine, one of the main components extracted from Sophora flavescens, has exhibited pharmacological effects on the differentiation in rat liver oval cells. However, its function and mechanism have not yet been fully elucidated. To further investigate them, an in vitro model was established using a rat liver oval cell line called WB‑F344 and treated with matrine. Initially, a significant increase in the number of monodansylcadaverine‑positive cells and in the levels of microtubule‑associated protein 1A/1B‑light chain 3 (LC3)‑II, which is a specific marker for detecting autophagy, was observed in matrine‑treated cells. This indicated that autophagy was stimulated by matrine, which was further confirmed by transmission electron microscopy. Additionally, the apoptotic oval cells were easily detected under matrine treatment using an Annexin‑V‑fluorescein isothiocyanate/propidium iodide assay, indicating that autophagy and apoptosis were synchronously induced by matrine. A decrease in B‑cell lymphoma (Bcl‑2) mRNA expression, but an increase in Bcl2‑associated X protein (Bax) mRNA expression were observed in matrine‑treated cells, which led to an upregulation of the Bax/Bcl‑2 ratio, a molecular marker for determining the extent of apoptosis. Next, the molecular mechanism of matrine‑induced autophagy and apoptosis was analyzed in WB‑F344 cells. β‑catenin degradation was downregulated by matrine and rapamycin, a foregone chemical agonist of autophagy, whereas it was upregulated by 3‑methyladenine, a specific inhibitor of autophagy. Additionally, β‑catenin activation induced an increase in LC3‑II levels and reversed the Bax/Bcl‑2 mRNA ratio under matrine treatment, whereas inhibition of β‑catenin by RNA interference induced a decrease of the LC3‑II amount and of the Bax/Bcl‑2 mRNA ratio. Finally, matrine treatment attenuated p53; however, with little or no change in LC3‑II levels, but a decrease in β‑catenin levels occurred in WB‑F344 cells upon treatment with pifithrin‑α, a chemical inhibitor of p53, revealing that p53, interfering with β‑catenin, may not be involved in matrine‑induced autophagy in WB‑F344 cells. These results demonstrate that β‑catenin is involved in matrine‑induced autophagy and apoptosis in WB‑F344 cells, while β‑catenin is negatively regulated by autophagy and positively by p53, indicating that β‑catenin may be involved in the crosstalk between autophagy and apoptosis in WB‑F344 cells.

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