Effect of Silencing LNK Gene on the Expression of EPO and EPOR in THP-1 Cells.
10.19746/j.cnki.issn.1009-2137.2019.04.004
- Author:
Run-Mei TIAN
1
;
Xi LUO
1
;
Mei TAN
1
;
Liang SHI
1
;
Cheng-Shuang HUANG
1
;
Qiong SU
1
;
Yan CHEN
2
Author Information
1. Second Department of Pediatrics, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China.
2. Second Department of Pediatrics, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China,E-mail: cyz600@ 163.com.
- Publication Type:Journal Article
- MeSH:
Blotting, Western;
Erythropoietin;
Gene Silencing;
Humans;
Proteins;
genetics;
Receptors, Erythropoietin;
THP-1 Cells
- From:
Journal of Experimental Hematology
2019;27(4):1013-1019
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of silencing LNK gene on the expression of EPO and EPOR in acute myeloid leukemia cells (THP-1).
METHODS:THP-1 cells were cultured. The lentivirus was used as a vector to silence the LNK gene stably. After 72 hours of infection, GFP expression level was detected by the fluorescent inverted microscopy. The lentiviral Infection efficiencies were monitored by flow cytometry. The LNK silencing effect was confirmed. The mRNA expressions of EPO and EPOR were detected by RT-PCR. The protein levels of LNK, EPO and EPOR were detected by Western blot.
RESULTS:At the time-point of 72 hours after lentivirus infection, the expression level of GFP was above 85% detected by fluorescent inverted microscopy. The infection efficiency was above 99% by flow cytometry. mRNA expressions of LNK, EPO and EPOR in LNK silencing group were signifycantly lower than those in control group (P＜0.05). The protein levels of LNK, EPO and EPOR in LNK silencing group were significantly lower than those in the control group (P＜0.05).
CONCLUSION:THP-1 cell line of LNK gene silencing has been successfully established,the LNK gene has been silenced, the expression of EPO and EPOR decrease, indicating that LNK may participate in the regulation of EPO and EPOR.
