Effect of siRNA targeting survivin gene on the biological behavior of hepatocellular carcinoma

Abstract

To study the influence of siRNA targeting survivin gene on the biological behavior of hepatocellular carcinoma (HCC), one pair of 21bp reverse repeated motifs of survivin target sequence with 9 spacers were synthesized and inserted into plasmid psilencer2. 1 to generate siRNA eukaryotic expression vector. After stable transfection into HepG2 cells, the biological behaviors of the survivin siRNA transfected HCC cells were observed. After the recombinant plasmid Psilence (+)-survivin was successfully constructed, survivin mRNA and protein expression inhibition ratio reached 73% and 75% respectively compared to control groups. Transfected cells with survivin siRNA demonstrated significantly inhibited cell growth and increased apoptosis. Subsequent study in nude mouse model demonstrated lower succeeding rate in cells transfected with survivin siRNA and slow growth rate. The results elucidated the siRNA targeting survivin gene could specially suppress its expression in HepG2 cells and inhibit tumor cells growth both in vivo and in vitro. This provides a theoretical basis to turn the drug resistance in tumor cells.

Keywords

hepatocellular carcinoma / siRNA / survivin / expression / inhibition

References

[1] GreenD R, EvanG I. A matter of life and death. Cancer cell, 2002, 1(1): 19-19

[2] ChangQ, LiuZ R, WangD Y, et al.. Survivin expression induced by doxorubicin in cholangiocarcinoma. World J Gastroenterol, 2004, 10(3): 415-415

[3] SongE, LeeS K, WangJ, et al.. RNA interference targeting Fas protects mice from fulminant hepatitis. Nat Med, 2003, 9(3): 347-347

[4] ChiouS K, JonesM K, TarnawskiA S. Survivin an anti-apoptosis protein: its biological roles and implications for cancer and beyond. Med Sci Monit, 2003, 9(4): 25-25

[5] BrantlS. Antisense-RNA regulation and RNA interference. Biochim Biophys Acta, 2002, 1575(1–3): 15-15

[6] AgrawalN, DasaradhiP V, MohmmedA, et al.. RNA interference: biology, mechanism, and applications. Microbiol Mol Biol Rev, 2003, 67(4): 657-657

[7] ScherrM, MorgenM A, EderM. Gene silencing mediated by small interfering RNAs in mammalian cell. Curr Med Chem, 2003, 10(3): 245-245

[8] CaplenN J. RNAi as a gene therapy approach. Expert Opin Biol Ther, 2003, 3(4): 575-575

[9] SuiG, SoohooC, AffarE I B, et al.. A DNA vector based RNAi technology to suppress gene expression in mammalian cells. Proc Nati Acad Sci USA, 2002, 99(8): 5515-5515

[10] ItoT, ShirakiK, SugimotoK, et al.. Survivin promotes cell proliferation in human hepatocellular carcinoma. Hepatology, 2000, 31(5): 1080-1080