[Suppression of SMYD3 expression in HepG2 cell by shRNA interference]

Jun-yao Xu, Li-bo Chen, Jun-yang Xu, Zhen Yang, Hai-yan Wei, Rong-hua Xu
Zhonghua Gan Zang Bing za Zhi, Zhonghua Ganzangbing Zazhi, Chinese Journal of Hepatology 2006, 14 (2): 105-8

OBJECTIVES: To identify the inhibition effect of shRNA on the SMYD3 (SET- and MYND-domain containing protein-3) expression in hepatoma cell line HepG2 through gene silencing.

METHODS: Two reverse repeated motifs targeting on the SMYD3 mRNA sequences 267-288, 302-323 respectively, were synthesized and inserted into the mock plasmid pGenesil-1 which expressed EGFP to create recombinant plasmids pGenesil-1-s1 and pGenesil-1-s2. pGenesil-1-hk specific to no SMYD3 mRNA sequence served as a control. After transfection into HepG2 cells, RT-PCR and western blot were applied to identify the down regulation of SMYD3 expression by shRNAs.

RESULTS: All plasmids were constructed successfully. pGenesil-1-s1, pGenesil-1-s2 inhibited the mRNA and protein expression of SMYD3 in HepG2 cells. There was a significant distinction when compared with pGenesil-1-hk and pGenesil-1 (P<0.01).

CONCLUSION: Short hairpin RNAs can efficiently and specifically suppress the expression of SMYD3 in HepG2 cells.

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