Suppression of SMYD3 expression in HepG2 cell by shRNA interference / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVES</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>Short hairpin RNAs can efficiently and specifically suppress the expression of SMYD3 in HepG2 cells.</p>