Sann-Joong-Kuey-Jian-Tang induces autophagy in HepG2 cells via regulation of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and p38 mitogen-activated protein kinase pathways

Chuang,Wan-Ling; Su,Chin-Cheng; Lin,Ping-Yi; Lin,Chi-Chen; Chen,Yao-Li

doi:10.3892/mmr.2015.3573

Sann-Joong-Kuey-Jian-Tang induces autophagy in HepG2 cells via regulation of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and p38 mitogen-activated protein kinase pathways

Authors:
- Wan-Ling Chuang
- Chin-Cheng Su
- Ping-Yi Lin
- Chi-Chen Lin
- Yao-Li Chen
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Affiliations: Transplant Medicine and Surgery Research Centre, Changhua Christian Hospital, Changhua 50006, Taiwan, R.O.C., Department of Surgery, Changhua Christian Hospital, Changhua 50006, Taiwan, R.O.C., Institute of Biomedical Science, National Chung-Hsing University, Taichung 40227, Taiwan, R.O.C.
Published online on: March 31, 2015 https://doi.org/10.3892/mmr.2015.3573
Pages: 1677-1684
Copyright: © Chuang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Sann-Joong-Kuey-Jian‑Tang (SJKJT), a traditional Chinese medicine, was previously reported to induce autophagy and inhibit the proliferation of the human HepG2 hepatocellular carcinoma cell line via an extrinsic pathway. In the present study, the effects of SJKJT‑induced autophagy and the cytotoxic mechanisms mediating these effects were investigated in HepG2 cells. The cytotoxicity of SJKJT in the HepG2 cells was evaluated using a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. The results demonstrated that the half‑maximal inhibitory concentration of SJKJT was 2.91 mg/ml at 24 h, 1.64 mg/ml at 48 h and 1.26 mg/ml at 72 h. The results of confocal fluorescence microscopy indicated that SJKJT resulted in the accumulation of green fluorescent protein‑LC3 and vacuolation of the cytoplasm. Flow cytometric analysis revealed the accumulation of acidic vesicular organelles. Furthermore, western blot analysis, used to determine the expression levels of autophagy‑associated proteins, demonstrated that the HepG2 cells treated with SJKJT exhibited LC3B‑Ⅰ/LC3B‑Ⅱ conversion, increased expression levels of Beclin, Atg‑3 and Atg‑5 and reduced expression levels of p62 and decreased signaling of the phosphoinositide‑3 kinase/Akt/mammalian target of rapamycin and the p38 mitogen‑activated protein kinase pathways. Taken together, these findings may assist in the development of novel chemotherapeutic agents for the treatment of malignant types of liver cancer.

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