Construction and identification of expressing siRNA plasmid against human augmenter of liver regeneration.
- Author:
Lin TANG
1
;
Qi LIU
;
Hang SUN
;
Ni TANG
;
Hui GUO
;
Jian-Chuan DENG
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Humans; Liver Neoplasms; genetics; metabolism; pathology; Molecular Sequence Data; Plasmids; genetics; Proteins; genetics; pharmacology; RNA, Small Interfering; genetics; Transfection
- From: Chinese Journal of Hepatology 2004;12(9):534-537
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESTo detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2 cells. To develop a kind of RNAi that specifically targets human augmenter of liver regeneration by synthesizing small interfering RNA (siRNA) in vivo, and to assess the inhibitory effect of this siRNA on hALR expression.
METHODSThe expression of hALR in HepG2 cells was observed with immunocytochemistry. The RNAi plasmid pSIALR-A and the unrelated control plasmid pSIALR-B were transfected into HepG2 cells. Forty-eight hours after transfection, the protein level of hALR was measured with immunocytochemistry; meanwhile, the reverse transcription PCR (RT-PCR) was performed to detect the expression of hALR mRNA.
RESULTShALR was expressed by HepG2 cells. siRNA plasmid pSIALR-A, which targets the cDNA of hALR and the unrelated control plasmid pSIALR-B, was successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly, compared with that of pSIALR-B.
CONCLUSIONThe results showed that the small interfering RNA targeting hALR suppresses the expression of hALR in a sequence-specific manner