Effect of Bmi-1 Expression on Chemotherapy Sensitivity in THP-1 Cells.
10.19746/j.cnki.issn.1009-2137.2021.02.009
- Author:
Si-Cong DONG
1
;
Ru-Nan JING
1
;
Hao PEI
1
;
Fan LIU
1
;
Bao-Xia ZHAO
1
;
Xiu-Xiang MENG
2
Author Information
1. Department of Clinical Hematology of Dalian Medical University, Dalian 116044, Liaoning Province, China.
2. Department of Clinical Hematology of Dalian Medical University, Dalian 116044, Liaoning Province, China,E-mail: xiuxiang_meng@sina.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Camptothecin/pharmacology*;
Cell Line, Tumor;
Cell Proliferation;
THP-1 Cells
- From:
Journal of Experimental Hematology
2021;29(2):363-368
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of Bmi-1 expression on the chemosensitivity of THP-1 cells and its relative mechanism.
METHODS:The pGenesil-2-Bmi-1 1 siRNA, p-MSCV-Bmi-1 plasmid was transfected into THP-1 cells to reduce or increase the expression of Bmi-1. The expression of Bmi-1 mRNA and protein was verified by PCR and Western blot. The effect of camptothecin (CPT) on the proliferation and chemosensitivity of THP-1 cells affected by Bmi-1 gene were detected by MTT assay. The expression of DNA double-strand breaks marker-γ-H2AX was detected by immunofluorescence assay. Mitochondrial membrane potential and apoptosis were observed by flow cytometry. The expression of Cytochrome C, Caspase 3, Bax and BCL-2 was detected by Western blot.
RESULTS:Silencing Bmi-1 could inhibit proliferation and enhance the sensitivity of THP-1 cells to CPT, while overexpressed Bmi-1 could promote the cell proliferation and attenucate sensitivity of THP-1 cells to CPT. Silencing Bmi-1 could enhance CPT-induced DNA double-strand breaks, decrease mitochondrial membrane potential and promote CPT-induced apoptosis. While increasing Bmi-1 gene expression could attenuate CPT-induced DNA double-strand breaks, enhamce mitochondrial membrane potential and significantly reduce CPT-induced apoptosis of cells.
CONCLUSION:Bmi-1 expression could influence the sensitivity of THP-1 cells to CPT, and its relative mechanism may relate to DNA double-strand breaks and endogenous apoptotic pathways.
