Biological Effects of the SARI Over-expression on K562 Cell Line.
- Author:
Ping WANG
1
;
Xiang-Jun CHEN
2
;
Jin-E ZHENG
2
;
Wen DU
2
;
Wei LIU
2
;
Jia-Hua ZHANG
2
;
Jun-Xia YAO
2
;
Xiao-Qing LI
2
;
Yan-Li HE
3
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Basic-Leucine Zipper Transcription Factors; Cell Cycle; Cell Line; Cell Proliferation; DNA, Complementary; Gene Expression; Genetic Vectors; Humans; K562 Cells; Lentivirus; Plasmids; Transfection; Tumor Suppressor Proteins
- From: Journal of Experimental Hematology 2015;23(3):637-641
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a lentivirus vector carrying SARI gene and to investigate its biological effects on K562 cells.
METHODSSARI was amplified from the plasmid containing SARI cDNA and subcloned into pLOV.CMV.eGFP virus vector. After sequencing, lentivirus packaging, titering, the viruses of SARI-pLOV.CMV.eGFP were harvested and tansfected into the K562 cells. Real-time quantitive PCR and Western blot were performed to validate the SARI expression at the level of mRNA and protein respectively. Simultaneously, the proliferation, apoptosis and cell cycle of K562 cells were detected by CCK-8 and flow cytometry respectively.
RESULTSThe SARI overexpressed lentivirus vector was successfully constructed. The mRNA and protein levels of SARI increased significantly in the pLOV.CMV.eGFP-SARI group, which was confirmed by Q-PCR and Western blot; as compared with blank and mock groups, SARI over-expression leaded to significant proliferation inhibition and increased apoptosis of K562 cells, without visible effects on cell cycle.
CONCLUSIONthe over-expression of SARI gene obviously suppresses the cell proliferation of the K562 cells as well as promotes the apoptosis. The results implied that the induction of the SARI gene expression may be an important candidate therapeutic method for the CML.
