Growth inhibition of K562 cells by cyclin E gene-specific small interfering RNA.
OBJECTIVE
To study the inhibitory effect of small interference RNA(siRNA) of cyclin E gene on the growth of K562 cells.


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 Lipofectamine2000. 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 G(1)-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.


CONCLUSIONS
RNA interference induces obvious inhibition of cyclin E gene expression, which consequently affects the proliferation of K562 cells.
Song
Hai-Xing
HX
Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China.
Ma
Wen-Li
WL
Song
Yan-Bin
YB
Zhang
Bao
B
Zheng
Wen-Ling
WL
D049109:Cell Proliferation; D019927:Cyclin E / Q000096:biosynthesis* / Q000235:genetics; D020868:Gene Silencing; D006801:Humans; D020014:K562 Cells; D015513:Oncogene Proteins / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
C475820:CCNE1 protein, human; D019927:Cyclin E; D015513:Oncogene Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering