Effects of beta-catenin gene silencing on stemness transcription factors in hepatocellular carcinoma stem cells

doi:10.12307/2023.364

Abstract

Abstract: BACKGROUND: Hepatocellular carcinoma stem cell is responsible for the formation, renewal and proliferation of cancer cells. It is a major topic to find key targets for hepatocellular carcinoma stem cell therapy and to further expand comprehensive treatment means for liver cancer.
OBJECTIVE: To observe changes of stemness transcription factors after β-catenin gene silencing on CD133+HepG2 cells, and investigate the action mechanism of β-catenin regulating stemness of hepatocellular carcinoma stem cells.
METHODS: After CD133+HepG2 hepatocellular carcinoma stem cells were screened by magnetic activated cell sorting, these cells were induced and cultured in stem cell culture medium. Positive rate of CD133+HepG2 hepatocellular carcinoma stem cells was identified by flow cytometry. β-Catenin shRNA lentiviral plasmid was constructed to inhibit β-catenin expression by RNA interference. After transfected CD133+HepG2 hepatocellular carcinoma stem cells for 72 hours, plasmid was labeled with fluorescence, which evaluated transfection efficiency. After β-catenin silencing was confirmed, CD133+HepG2 hepatocellular carcinoma stem cell proliferation was measured by plate clone formation assay. CD133+HepG2 hepatocellular carcinoma stem cell cycle was detected by flow cytometry. RT-PCR and western blot assay were used to detect the mRNA and protein expression of stemness transcription factors SOX2, NANOG and OCT4.
RESULTS AND CONCLUSION: Compared with blank control group and virus-free group, clone formation ability was decreased in virus-blockade group. Percentage of G0/G1 phase cells increased significantly, while that of S phase cells and G2/M phase cells decreased. Protein and mRNA expression of NANOG, SOX2 and OCT4 was significantly decreased in virus-blockade group (P < 0.05). It is indicated that down-regulation of β-catenin expression may have a potential effect on suppressing the stemness of hepatocellular carcinoma stem cells.

Key words: hepatocellular carcinoma stem cell, stem cell transcription factor, β-catenin, RNA interference, CD133, HepG2

CLC Number:

Sun Hua, Qin Yanchun, Rong Zhen, Li Zulong, Jiang Ruiyuan, Zhong Xiaoting, Mo Chunmei. Effects of beta-catenin gene silencing on stemness transcription factors in hepatocellular carcinoma stem cells[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(15): 2297-2303.

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