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Effect of Bmi-1-mediated NF-κB signaling pathway on the stem-like properties of CD133+ human liver cancer cells
- Source :
- Cancer Biomarkers. 22:575-585
- Publication Year :
- 2018
- Publisher :
- IOS Press, 2018.
-
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.
- Subjects :
- 0301 basic medicine
Homeobox protein NANOG
Cancer Research
Cell
Fluorescent Antibody Technique
Gene Expression
Apoptosis
Flow cytometry
03 medical and health sciences
0302 clinical medicine
SOX2
Cell Line, Tumor
Genetics
medicine
Humans
AC133 Antigen
neoplasms
Polycomb Repressive Complex 1
medicine.diagnostic_test
Chemistry
Cell Cycle
Liver Neoplasms
NF-kappa B
General Medicine
Cell cycle
Cell biology
Gene Expression Regulation, Neoplastic
030104 developmental biology
medicine.anatomical_structure
Oncology
030220 oncology & carcinogenesis
embryonic structures
Neoplastic Stem Cells
biological phenomena, cell phenomena, and immunity
Stem cell
Signal transduction
Signal Transduction
Subjects
Details
- ISSN :
- 18758592 and 15740153
- Volume :
- 22
- Database :
- OpenAIRE
- Journal :
- Cancer Biomarkers
- Accession number :
- edsair.doi.dedup.....f1eac962543cc0bb4cf9c6e7aeaadba2