Suppression of SMYD3 expression in HepG2 cell by shRNA interference.
- Author:
Jun-yao XU
1
;
Li-bo CHEN
;
Jun-yang XU
;
Zhen YANG
;
Hai-yan WEI
;
Rong-hua XU
Author Information
- Publication Type:Journal Article
- MeSH: Carcinoma, Hepatocellular; metabolism; pathology; Cell Line, Tumor; Down-Regulation; Histone-Lysine N-Methyltransferase; biosynthesis; genetics; Humans; Liver Neoplasms; metabolism; pathology; RNA Interference; RNA, Messenger; biosynthesis; genetics; RNA, Small Interfering; genetics; Transfection
- From: Chinese Journal of Hepatology 2006;14(2):105-108
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESTo 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.
METHODSTwo 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.
RESULTSAll 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).
CONCLUSIONShort hairpin RNAs can efficiently and specifically suppress the expression of SMYD3 in HepG2 cells.
