Kangxianruangan granule‑containing serum mediated inhibition of hepatic oval cell differentiation into hepatocellular carcinoma cells via the Wnt‑1/β‑catenin signaling pathway

Tang,Wenqian; Xue,Juan; Luo,Lei; Wang,Yao; Cai,Xin; Liu,Yuqing; Huang,Dawei; Wang,Xiaodong; He,Tangqing; Lu,Dingbo; Yang,Fan

doi:10.3892/mmr.2021.12571

Kangxianruangan granule‑containing serum mediated inhibition of hepatic oval cell differentiation into hepatocellular carcinoma cells via the Wnt‑1/β‑catenin signaling pathway

Authors:
- Wenqian Tang
- Juan Xue
- Lei Luo
- Yao Wang
- Xin Cai
- Yuqing Liu
- Dawei Huang
- Xiaodong Wang
- Tangqing He
- Dingbo Lu
- Fan Yang
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Affiliations: Health Management Centre, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, Hubei 430074, P.R. China, Clinical College of Chinese Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430061, P.R. China, Department of Infectious Diseases, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Infectious Diseases, The People's Hospital of Jiangshan, Jiangshan, Zhejiang 324100, P.R. China, Department of Infectious Diseases, Yancheng TCM Hospital Affiliated Nanjing University of Chinese Medicine, Nanjing, Jiangsu 224001, P.R. China, Department of Hepatology, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, Hubei 430074, P.R. China
Published online on: December 15, 2021 https://doi.org/10.3892/mmr.2021.12571
Article Number: 55
Copyright: © Tang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hepatocellular carcinoma is a malignancy with poor clinical prognosis. Hepatic oval cells (HOCs) tend to differentiate into cancerous hepatocellular carcinoma cells (HCCs) in the tumor microenvironment. The purpose of the present study was to explore the role of kangxianruangan granule (KXRG)‑containing serum in inhibiting the differentiation of HOCs into HCCs via the Wnt‑1/β‑catenin signaling pathway. N‑methyl‑N'‑nitro‑N‑nitrosoguanidine (MNNG) was applied to induce the transformation of the rat HOC cell line WB‑F344 into HCCs. The overexpression plasmid, Wnt‑1‑up, was utilized to increase Wnt‑1 expression. Subsequently, high, medium and low concentrations of KXRG were applied to MNNG‑treated WB‑F344 cells to assess the inhibitory effect of KXRG on cell differentiation. Flow cytometry was conducted to detect the cell cycle distribution, apoptotic rate and expression of cytokeratin‑19 (CK‑19) protein in cells. An immunofluorescence double staining protocol was used to detect the expression of Wnt‑1 and β‑catenin. ELISAs were performed to detect α fetoprotein in the cell supernatants. Reverse transcription‑quantitative PCR and western blotting were conducted to detect the mRNA and protein expression levels of Wnt‑1, β‑catenin, Cyclin D1, C‑myc, matrix metalloproteinase‑7 (MMP‑7), Axin2 and epithelial cell adhesion molecule (EpCAM) in cells. Compared with the normal group, the apoptotic rate, proportion of S phase cells, concentration of AFP in the cell supernatant, level of CK‑19 protein, and mRNA and protein expression levels of Wnt‑1, β‑catenin, Cyclin D1, C‑myc, MMP‑7, Axin2 and EpCAM were all significantly increased in the model group. Addition of KXRG significantly reduced the aforementioned indicators compared with the model group. Moreover, Wnt‑1 overexpression further increased the aforementioned indicators compared with the model group, whereas KXRG significantly inhibited these effects. The results indicated that KXRG inhibited the differentiation of HOCs into HCCs via the Wnt‑1/β‑catenin signaling pathway, which suggested the potential clinical application of KXRG for the prevention of hepatocellular carcinoma.

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