SPARC Overexpression Enhances the Sensitivity of SKM-1 Cells to Ara-C by Regulating CPBP/MLKL.
10.19746/j.cnki.issn.1009-2137.2022.05.031
- Author:
Si-Min LIANG
1
;
Xiao-Jia ZHOU
1
;
Duo CAI
1
;
Qiao ZHOU
1
;
Li WANG
2
Author Information
1. Department of Hematology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
2. Department of Hematology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China. E-mail: liwangls@yahoo.com.
- Publication Type:Journal Article
- Keywords:
CPBP;
SKM-1 cells;
SPARC gene;
cytarabine arabinoside;
sensitivity
- MeSH:
Apoptosis;
Cell Line, Tumor;
Cell Proliferation;
Cytarabine;
Humans;
Kruppel-Like Factor 6/metabolism*;
Osteonectin/pharmacology*;
Protein Kinases/pharmacology*;
Proto-Oncogene Proteins c-akt/metabolism*;
Proto-Oncogene Proteins c-bcl-2/metabolism*;
RNA, Messenger;
Sincalide/pharmacology*;
Tumor Suppressor Protein p53;
bcl-2-Associated X Protein/pharmacology*
- From:
Journal of Experimental Hematology
2022;30(5):1508-1514
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of SPARC gene overexpression on the chemotherapeutic sensitivity of AML-MDS cell line SKM-1 to Ara-C and to further explore its mechanism.
METHODS:Subjects were divided into 6 groups: SKM-1 cells (Control), Negative control (LV-NC), SPARC overexpression (LV-SPARC), SKM-1 cells+30 ng/ml Ara-C (30 ng/ml Ara-C), LV-NC+30 ng/ml Ara-C and LV-SPARC+30 ng/ml Ara-C. Cell activity was detected by CCK-8 assay, cell cycle distribution and apoptosis were detected by flow cytometry, mRNA expression levels of SPARC, CPBP and MLKL were detected by RT-qPCR, and the expression levels of related protein were detected by Western blot.
RESULTS:After co-treatment with SPARC overexpression and Ara-C, the cell viability decreased and apoptosis increased significantly, with obvious up-regulation of Bax and down-regulation of BCL-2 (P<0.05). Compared with the control group, the cell cycle of LV-SPARC+30 ng/ml Ara-C group was significantly arrested in S phase with obvious down-regulation of CDK2 and up-regulation of p27KIP1 (P<0.05). Compared with LV-SPARC group and 30 ng/ml Ara-C group, the mRNA and protein expression levels of CPBP and MLKL (p-MLKL) were significantly elevated in LV-SPARC+30 ng/ml Ara-C group (P<0.05). In addition, after co-treatment with SPARC overexpression and Ara-C, the protein expression level of p-AKT decreased and the protein expression level of p53 increased (P<0.05).
CONCLUSION:SPARC overexpression enhanced the sensitivity of SKM-1 cells to Ara-C and promoted cell cycle arrest and apoptosis, the mechanism of which may be related to the regulation of CPBP/MLKL pathway.
