Effect of miR-203/CREB1 Signaling Regulation Mediated by DNA Methylation on the Proliferation and Apoptosis of Multiple Myeloma Cells.
10.19746/j.cnki.issn.1009-2137.2022.03.021
- Author:
Cheng-Bo XU
1
;
Bin LIAO
1
;
Hai-Ying FU
2
;
Yan QI
1
;
Jian-Zhen SHEN
3
Author Information
1. Department of Hematology, The Affiliated People's Hospital to Fujian University of Traditional Chinese Medicine, Fuzhou 350004, Fujian Province, China.
2. Department of Hematology, Fujian Medical University Union Hospital, Fujian Institute of Hematology, Fuzhou 350001, Fujian Province, China.
3. Department of Hematology, Fujian Medical University Union Hospital, Fujian Institute of Hematology, Fuzhou 350001, Fujian Province, China,E-mail: albertsky@163.com.
- Publication Type:Journal Article
- Keywords:
CREB1;
apoptosis;
methylation;
miR-203;
multiple myeloma
- MeSH:
Apoptosis;
Azacitidine/pharmacology*;
Cell Line, Tumor;
Cell Proliferation;
Cyclic AMP Response Element-Binding Protein/pharmacology*;
DNA Methylation;
Gene Expression Regulation, Neoplastic;
Humans;
MicroRNAs/metabolism*;
Multiple Myeloma/genetics*;
RNA, Messenger/metabolism*
- From:
Journal of Experimental Hematology
2022;30(3):790-796
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of miR-203/CREB1 signaling regulation mediated by DNA methylation on the proliferation, invasion and apoptosis of multiple myeloma (MM) cells.
METHODS:The methylation level of miR-203 in the RPMI 8226 cells was detected by bisulfite sequcucing polymerase chain reaction (BSP). The mRNA expression of miR-203 was measured by quantitative real-time polymerase chain reaction. RPMI 8226 cells were treated with DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR). The miR-203 mimic in MM cell line RPMI 8226 was transfected to establish overexpressed miR-203 cell. The proliferation, invasion ability and apoptosis of RPMI 8226 cell was detected by CCK-8 assay, Transwell, and flow cytometry, respectively. The targeting relationship between miR-203 and CREB1 was verified by double luciferase report assay. Western blot was used to detect the expression of CREB1 protein.
RESULTS:Hypermethylation of miR-203 promoter region and low expression level of miR-203 mRNA were detected in the RPMI 8226 cells, which showed that demethylation could induce the expression of miR-203. The proliferation and invasion ability of RPMI 8226 cells after treated by 5-Aza-CdR were inhibited, and showed statistical significance as compared with blank control group (both P<0.05)，while the apoptosis rate was promoted (P<0.05). The proliferation, invasion ability and apoptosis of overexpressed miR-203 were the same as the demethylation group. Double luciferase report assay confirmed that CREB1 was the direct target of miR-203. The protein level of CREB1 was inhibited by demethylation and showed statistical significance as compared with control group (P<0.05).
CONCLUSION:MiR-203 targeting CREB1 mediated by DNA methylation leads to maintain the malignant biological behaviors of MM cells.
