[Construction and identification of expressing siRNA plasmid against human augmenter of liver regeneration].
OBJECTIVE
To 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.


METHODS
The 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.


RESULTS
hALR 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.


CONCLUSIONS
The results showed that the small interfering RNA targeting hALR suppresses the expression of hALR in a sequence-specific manner
Tang
Lin
L
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Liu
Qi
Q
Sun
Hang
H
Tang
Ni
N
Guo
Hui
H
Deng
Jian-Chuan
JC
D001483:Base Sequence; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000235:genetics* / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D050862:Oxidoreductases Acting on Sulfur Group Donors
D011506:Proteins; D034741:RNA, Small Interfering; C088825:GFER protein, rat; D050862:Oxidoreductases Acting on Sulfur Group Donors