Huaier inhibits proliferation and metastasis of liver cancer cells by inducing autophagy

doi:10.3877/cma.j.issn.2095-3232.2019.02.019

Abstract

Abstract:

Objective

To evaluate the effect of Huaier on the proliferation and metastasis of human hepatocellular carcinoma (HCC) cells, and to explore the molecular mechanism.

Methods

HCC cell lines SK-HEP-1 and HEP3B were divided into the control, Huaier treatment and autophagy inhibitor groups. The cell viability was detected by CCK-8 assay. The cell proliferation was determined by clone formation assay. The cell apoptosis was detected by flow cytometry. The cell metastasis was detected by Transwell assay. The formation of autophagosome was observed by fluorescent GFP-LC3 fusion protein. The related proteins of epithelial-mesenchymal transition (EMT), autophagy and Akt/mTOR signaling pathway were detected by Western blot. Comparison between 2 groups was conducted by t test. Multi-group comparison was carried out by One-way ANOVA and LSD-t test.

Results

The activity of SK-HEP-1 and HEP3B decreased with the increase of Huaier concentration and the prolongation of treatment time. In 6 g/L Huaier treatment group, the number of clone formation of SK-HEP-1 and HEP3B cells was 77±9 and 84±9 respectively, significantly less than 231±15 and 257±35 in control group (t=-8.90, -6.33; P<0.05). Huaier significantly promoted the cell apoptosis in a concentration-dependent manner (F=120.88, 199.59; P<0.05). In 8 g/L Huaier treatment group, the number of trans-membrane SK-HEP-1 and HEP3B cells was 68±13 and 54±13, significantly less than 389±28 and 411±15 in control group (t=-17.98, -31.47; P<0.05). Western blot demonstrated that the expression of N-cadherin and Vimentin proteins was down-regulated, LC3Ⅱ expression was up-regulated and LC3Ⅰ expression was down-regulated in the Huaier treatment group. The expression levels of p-Akt, p-mTOR and p62 proteins were down-regulated and the expression of LC3Ⅱ was up-regulated. Autophagy inhibitor 3-MA could reduce the autophagy in Huaier treatment group.

Conclusions

Huaier exerts anti-tumor effect by promoting the apoptosis and inhibiting the proliferation and metastasis of HCC cells. The mechanism is probably related to suppressing the Akt/mTOR signaling pathway, inducing autophagy and preventing EMT.

Key words: Carcinoma, hepatocellular, Autophagy, Signaling pathway, Cell proliferation, Cell movement

Wenxin Li, Xicheng Wang, Meihuan Xiang, Wenliang Tan, Yunxiuxiu Xu, Yibiao Ye, Jie Chen, Tao Chen. Huaier inhibits proliferation and metastasis of liver cancer cells by inducing autophagy[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2019, 08(02): 169-174.

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References 20

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