Effect of shRNA targeted to beta-catenin on K562 cell growth.
- Author:
Zeng-Jun LI
1
;
Qian LI
;
Guo-Rong WANG
;
Zhen YU
;
Chang-Hong LI
;
Ya-Fei WANG
;
Ye-Nan LI
;
Lu-Gui QIU
Author Information
1. Chinese Academy of Medical Science, Peking Union Medical College, Tianjin 300020, China.
- Publication Type:Journal Article
- MeSH:
Cell Proliferation;
Humans;
K562 Cells;
Neoplastic Stem Cells;
Plasmids;
genetics;
RNA Interference;
RNA, Messenger;
genetics;
metabolism;
RNA, Small Interfering;
genetics;
Transfection;
beta Catenin;
genetics;
metabolism
- From:
Journal of Experimental Hematology
2008;16(4):725-729
- CountryChina
- Language:Chinese
-
Abstract:
In order to investigate the effect of shRNA targeted to beta-catenin on the growth of K562 cells, plasmid containing beta-catenin specific shRNA sequence was transfected into K562 cells by lipofectamine 2000, and G418 was added to screen the positive cells. Real-time PCR and Western blot were used to detect the expression of beta-catenin. Cell growth curve, MTT and colony forming cell assays were used to evaluate the proliferation potential of cells. The results showed that the mRNA level of beta-catenin was reduced significantly in K562 cells transfected into interfering plasmid as compared with control plasmid, while the protein level failed to demonstrate difference by the time of 72 hours after transfection. After long-term culture with G418, the count of positive cells enhanced in control group while no positive cells survived in the interfering group. Colony-forming cell assays revealed that the K562 cells in interfering group formed colonies with very small size and low forming rate, compared with the control group, though the growth curve and MTT failed to illustrate differences. It is concluded that the beta-catenin-specific shRNA mediated by plasmid can effectively knockdown the expression of beta-catenin gene and inhibit the colony-forming ability in K562 cells, it is a potential target for the therapy of CML, even in blast crisis.
