Growth inhibition of K562 cells by cyclin E gene-specific small interfering RNA
10.3321/j.issn:1673-4254.2005.04.001
- VernacularTitle:cyclin E基因siRNA抑制K562细胞生长的研究
- Author:
Hai-Xing SONG
1
;
Wen-Li MA
;
Yan-Bin SONG
;
Bao ZHANG
;
Wen-Ling ZHENG
Author Information
1. 南方医科大学
- Keywords:
cyclin E;
RNA interference;
small interfering RNA;
K562 cells
- From:
Journal of Southern Medical University
2005;25(4):361-365
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the inhibitory effect of small interference RNA(siRNA) ofcyclin E gene on the growth ofK562cells. Methods siRNA targeting the 940 bp site of the cyclin E mRNA were designed and generated by PCR amplification.The PCR products containing U6 promoter and the siRNA were then transfected into K562 cells via LipofectamineTM2000.The cells transfected with non-functional siRNA served as the negative control group and those only treated with serum-free RPMI1640 as the blank control group. Cell counting, reverse transcriptase (RT)-PCR and flow cytometry were employed to evaluate the effect of RNA interference. Results Compared with the negative and blank control groups, the viable cell count in the interference group was decreased by approximately 80%, the ratio of G1-phase cells increased by nearly 30%, and growth arrest was observed. Cyclin E mRNA expression in the cells of the interference group was significantly lowered by about 70%as compared with that of the negative and blank control groups, whereas the latter two groups had similar expression levels.Conclusion RNA interference induces obvious inhibition of cyclin E gene expression, which consequently affects the proliferation ofK562 cells.
