Journal of Gastrointestinal & Digestive System
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To construct a short hairpin RNA ( shRNA ) eukaryotic expression vector targeting Livin or Survivin gene, and
explore the impact of Co-transfection of Livin and Survivin shRNA expression vector on biological behavior of HepG2 cells.
shRNA eukaryotic expression vector pSD11-Livin and pSD11-Survivin were designed, constructed, and transfected
into HepG2 cells in combination with liposome. Cells were divided into blank control group, negative control group, Survivin
group, Livin group and co-transfection group. mRNA relative expression was detected by fluorescence quantitative PCR. Western
blot were used to detect Livin, Survivin protein respectively. Changes of cell proliferation were detected by MTT, and TUNEL was
used to detect apoptosis rate.
The Livin and Survivin shRNA eukaryotic expression vectors were successfully constructed. Livin, Survivin mRNA
relative expression quantity in HepG2 cells of co-transfection group was 0.120? 0.022 and 0.325 ?0.125 respectively. Compared
with Survivin group or Livin group, mRNA relative expression quantity of co-transfection group was decreased significantly (P
< 0.05). Livin, Survivin protein relative expression quantity in HepG2 cells of co-transfection group was 0.412? 0.099 and 0.473
?0.051. Co-transfection inhibited protein expression more efficiently than single-transfection (P < 0.05). Cell growth inhibition
rate in co-transfection group were higher than those in single-transfection group on 48h, 60h, 72h after transfection (P < 0.05).
Apoptosis rate increased more significantly in co-transfection group than any other groups (P < 0.05).
Livin, Survivin shRNA eukaryotic expression vector was successfully constructed. Co-transfection of pSD11-Livin
and pSD11-Survivin reduced the expression of Livin and Survivin gene in HepG2 cells more effectively, inhibited the proliferation
of hepatoma cells more significantly, and induced the apoptosis of HepG2 cells more effectively.
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