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

• Published in: Cancer biomarkers : section A of Disease markers Vol. 22; no. 3; pp. 575 - 585
• Main Authors: Ma, De-Qiang, Zhang, Yin-Hua, Ding, De-Ping, Li, Juan, Chen, Lin-Li, Tian, You-You, Ao, Kang-Jian
• Format: Journal Article
• Language: English
• Published: London, England SAGE Publications 01.01.2018; Sage Publications Ltd
• Subjects: Apoptosis; Biological properties; Bmi protein; Cancer; Cell cycle; Cell migration; Colonies; Flow cytometry; Hepatocytes; Immunofluorescence; Liver; Liver cancer; NF-κB protein; Nuclear transport; Oct-4 protein; Programmable logic controllers; Signal transduction; Signaling; siRNA; Stem cells; Translocation; Western blotting; Liver cancer stem cells; Bmi-1; NF-κB pathway; CD133
• ISSN: 1574-0153; 1875-8592; 1875-8592
• DOI: 10.3233/CBM-181329

OBJECTIVE: To investigate the impact of Bmi-1-mediated NF- κ B 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- κ B 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- κ B p65 nuclear translocation, and qRT-PCR and Western blotting for the determination of SOX2, NANOG, OCT4, Bmi-1, and NF- κ B 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- κ B p65, the nuclear translocation of NF- κ B 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- κ B signaling pathway, which lays a scientific foundation for the clinical treatment of liver cancer.