Effect of Bmi-1-mediated NF-κB signaling pathway on the stem-like properties of CD133+ human liver cancer cells

Article type: Research Article

Authors: Ma, De-Qiang^{a; 1; *} | Zhang, Yin-Hua^{a; 1} | Ding, De-Ping^a | Li, Juan^b | Chen, Lin-Li^a | Tian, You-You^a | Ao, Kang-Jian^a

Affiliations: [a] Department of Infectious Diseases, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China | [b] Maternal and Child Health-Care Hospital, Shiyan, Hubei 442000, China

Correspondence: [*] Corresponding author: De-Qiang Ma, Department of Infectious Diseases, Taihe Hospital, Hubei University of Medicine, No. 32, South Renmin Road, Shiyan, Hubei 442000, China. Tel.: +86 13687205678; E-mail: [email protected].

Note: [1] These authors contributed equally to this study and share first authorship.

Abstract: OBJECTIVE: To investigate the impact of Bmi-1-mediated NF-κB pathway on the biological characteristics of CD133+ liver cancer stem cells (LCSCs). METHODS: Flow cytometry was used to isolate CD133+ LCSC cells from Huh7, Hep3B, SK-hep1, and PLC/PRF-5 cells. CD133+ Huh7 cells were divided into Control, Blank, Bmi-1 siRNA, JSH-23 (NF-κB pathway inhibitor), and Bmi-1 + JSH-23 groups. The properties of CD133+ Huh7 cells were detected by the colony-formation and sphere-forming assays. Besides, Transwell assay was applied for the measurement of cell invasion and migration, immunofluorescence staining for the detection of NF-κB p65 nuclear translocation, and qRT-PCR and Western blotting for the determination of SOX2, NANOG, OCT4, Bmi-1, and NF-κB p65 expression. RESULTS: CD133+ Huh-7 cells were chosen as the experiment subjects after flow cytometry. Compared with CD133- Huh-7 cells, the expression of CD133, OCT4, SOX2, NANOG, Bmi-1, and NF-κB p65, the nuclear translocation of NF-κB p65, the number of cell colonies and Sphere formation, as well as the abilities of invasion and migration were observed to be increased in CD133+ Huh-7 cells, which was inhibited after treated with Bmi-1 siRNA or JSH-23, meanwhile, the cell cycle was arrested at the G0/G1 and S phases with apparently enhanced cell apoptosis. Importantly, no significant differences in the biological characteristics of CD133 + Huh-7 cells were found between the Blank group and Bmi-1 + JSH-23 group. CONCLUSION: Down-regulating Bmi-1 may inhibit the biological properties of CD133+ LCSC by blocking NF-κB signaling pathway, which lays a scientific foundation for the clinical treatment of liver cancer.

Keywords: Liver cancer stem cells, Bmi-1, NF-κB pathway, CD133+

DOI: 10.3233/CBM-181329

Journal: Cancer Biomarkers, vol. 22, no. 3, pp. 575-585, 2018