Experimental research of therapeutic effect on hepatocellular carcinoma of targeting SMYD3 gene inhibition by RNA interference.
- Author:
Jun-yao XU
1
;
Li-bo CHEN
;
Jun-yang XU
;
Zhen YANG
;
Rong-hua XU
;
Hai-yan WEI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis; Carcinoma, Hepatocellular; genetics; pathology; therapy; Cell Line, Tumor; Genetic Therapy; methods; Histone-Lysine N-Methyltransferase; biosynthesis; genetics; Humans; Liver Neoplasms; genetics; pathology; therapy; Mice; Mice, Inbred BALB C; Mice, Nude; Plasmids; genetics; RNA Interference; RNA, Small Interfering; genetics; Transfection
- From: Chinese Journal of Surgery 2006;44(7):481-484
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo determine the potential of SMYD3 as a therapeutic target for hepatocellular carcinoma (HCC) by potent and highly sequence-specific RNA interference (RNAi) technique.
METHODSThe mRNA of SMYD3 was detected by RT-PCR in different HCC cell lines, such as HepG2, Hep3B and SMMC7721. Recombinant SMYD3 shRNA plasmid Pgenesil-1-s was constructed and transfected into HepG2 cells, and Western blot was used to identify the down regulation of SMYD3 protein expression after transfection. MTT and flow cytometry analysis (FCM) were respectively applied to analysis cell proliferation and apoptosis. In vivo study was carried out by injecting recombinant SMYD3 shRNA plasmids into transplanted tumors of nude mice.
RESULTSThe expression of SMYD3 mRNA was abundant in HCC cell lines HepG2, Hep3B, SMMC7721, whereas none in normal hepatic cell line L-02. RNA interference was able to suppress SMYD3 expression greatly and then inhibited cell growth effectively and induced apoptosis of HepG2 cells efficiently. After injection of recombinant SMYD3 shRNA plasmid, transplanted tumors grew slowly and reduced in size and weight when compared with those of control groups (P < 0.01).
CONCLUSIONSSMYD3 plays a major role in occurrence and progress of HCC. Inhibition of SMYD3 by RNAi can induce apoptosis in HepG2 cells and suppress tumor growth in nude mice. Therefore SMYD3 could be an ideal therapeutic target for HCC.
