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Growth inhibition of K562 cells by cyclin E gene-specific small interfering RNA
Hai-Xing Song1, Wen-Li Ma, Yan-Bin Song
1Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China.
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.
Conclusion:
RNA interference induces obvious inhibition of cyclin E gene expression, which consequently affects the proliferation of K562 cells.
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