Establishment of Drug-Resistant Cell Lines of Acute Myeloid Leukemia and Correlation of Sirt1 and PGC-1α Expression Levels with Drug Resistance.
10.19746/j.cnki.issn.1009-2137.2022.03.007
- Author:
Li-Cheng LI
1
;
Jun-Di WANG
1
;
Si-Si YANG
1
;
Zhen ZHOU
2
;
Qing-Fan ZENG
3
;
Fang ZHENG
4
Author Information
1. Department of Hematology, Baiyun Hospital Affiliated to Guizhou Medical University, Guiyang 550014, Guizhou Province, China.
2. Department of Pharmacy, Baiyun Hospital Affiliated to Guizhou Medical University, Guiyang 550014, Guizhou Province, China.
3. Department of Anesthesiology, Baiyun Hospital Affiliated to Guizhou Medical University, Guiyang 550014, Guizhou Province, China.
4. Department of Hematology, Baiyun Hospital Affiliated to Guizhou Medical University, Guiyang 550014, Guizhou Province, China,E-mail: 1179556708@qq.com.
- Publication Type:Journal Article
- Keywords:
PGC-1α;
Sirt1;
acute myeloid leukemia;
drug resistant
- MeSH:
Cell Line;
Cytarabine/pharmacology*;
Drug Resistance;
Humans;
Leukemia, Myeloid, Acute/genetics*;
RNA, Messenger/genetics*;
Sirtuin 1
- From:
Journal of Experimental Hematology
2022;30(3):704-710
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct cytarabine-resistant acute myeloid leukemia (AML) cell lines, and explore the correlation between Sirt1, PGC-1α expression levels and drug resistance.
METHODS:Human acute promyelocytic leukemia Kasumi-1 cells were induced by the method of gradually increasing the concentration of Ara-C drug. The IC50 value of Kasumi-1 cells before and after drug addition was detected by CCK-8 method, so as to construct Ara-C resistant cell lines. The expression levels of Sirt1 and PGC-1α mRNA in Kasumi-1 drug-resistant cell lines and their parental cell lines were detected by real-time fluorescence quantitative PCR, and the expression levels of Sirt1 and PGC-1α protein in kasumi-1 drug-resistant cell lines and their parental cell lines were detected by Western blot.
RESULTS:The constructed Kasumi-1 cell line had common morphological characteristics of drug-resistant cell lines under microscope, and the drug resistance index was greater than 5, indicating that Kasumi-1 drug-resistant cells had good drug resistance after the construction. The RT-qPCR and Western blot assays showed that the expression levels of Sirt1 and PGC-1α mRNA and protein in the drug-resistant cell lines were higher than those of the parental cell lines (P<0.001).
CONCLUSION:AML cell lines resistant to Ara-C can be successfully induced by the method of gradually increasing the concentration, and the co-high expression of Sirt1 and PGC-1α may mediate the drug resistance of AML cells.
