Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Survivin knockdown by short hairpin RNA abrogates the growth of human hepatocellular carcinoma xenografts in nude mice

Cancer Gene Therapy volume 17, pages 275–288 (2010)Cite this article

Abstract

Abnormal high activation of survivin is involved in carcinogenesis of various types of cancer. Survivin has been shown to promote cell proliferation in human hepatocellular carcinoma (HCC). Survivin-targeting approaches have become a promising strategy for treating HCC. Here, we used a reporter system to screen effective survivin siRNA sequences. The effect of vector-based survivin short hairpin RNA (shRNA) on the malignant phenotype of HCC cells in vitro and in vivo was determined, and an adenovirus-mediated shRNA expression vector was developed to decrease survivin expression of the established HCC tumor in nude mice. In vitro study showed that stable survivin knockdown inhibited cancer cell proliferation, enhanced apoptotic susceptibility, arrested cell cycle in the G1 phase and resulted in apparent mitotic catastrophe. Moreover, cells stably expressing survivin shRNA showed decreased tumorigenicity in nude mice. An additional in vivo study showed that intratumoral injection of adenovirus-delivered survivin shRNA suppressed tumor growth by spontaneous apoptosis of cancer cells and significantly prolonged animal survival. In conclusion, we proved the therapeutic potential of survivin shRNA for the treatment of HCC. And our results indicated that adenovirus-delivered shRNA may serve as a novel therapeutic for HCC.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Ambrosini G, Adida C, Altieri DC . A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 1997; 3: 917–921.

    Article  CAS  Google Scholar 

  2. Ambrosini G, Adida C, Sirugo G, Altieri DC . Induction of apoptosis and inhibition of cell proliferation by survivin gene targeting. J Biol Chem 1998; 273: 11177–11182.

    Article  CAS  Google Scholar 

  3. Li F, Ambrosini G, Chu EY, Plescia J, Tognin S, Marchisio PC et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 1998; 396: 580–584.

    Article  CAS  Google Scholar 

  4. Li F, Ackermann EJ, Bennett CF, Rothermel AL, Plescia J, Tognin S et al. Pleiotropic cell-division defects and apoptosis induced by interference with survivin function. Nat Cell Biol 1999; 1: 461–466.

    Article  CAS  Google Scholar 

  5. Pennati M, Colella G, Folini M, Citti L, Daidone MG, Zaffaroni N . Ribozyme-mediated attenuation of survivin expression sensitizes human melanoma cells to cisplatin-induced apoptosis. J Clin Invest 2002; 109: 285–286.

    Article  CAS  Google Scholar 

  6. Uchida H, Tanaka T, Sasaki K, Kato K, Dehari H, Ito Y et al. Adenovirus-mediated transfer of siRNA against survivin induced apoptosis and attenuated tumor cell growth in vitro and in vivo. Mol Ther 2004; 10: 162–171.

    Article  CAS  Google Scholar 

  7. Mesri M, Wall NR, Li J, Kim RW, Altieri DC . Cancer gene therapy using a survivin mutant adenovirus. J Clin Invest 2001; 108: 981–990.

    Article  CAS  Google Scholar 

  8. Grossman D, McNiff JM, Li F, Altieri DC . Expression and targeting of the apoptosis inhibitor, survivin, in human melanoma. J Invest Dermatol 1999; 113: 1076–1081.

    Article  CAS  Google Scholar 

  9. Tu SP, Jiang XH, Lin MC, Cui JT, Yang Y, Lum CT . Suppression of survivin expression inhibits in vivo tumorigenicity and angiogenesis in gastric cancer. Cancer Res 2003; 63: 7724–7732.

    CAS  PubMed  Google Scholar 

  10. Sun Y, Lin R, Dai J, Jin D, Wang SQ . Suppression of tumor growth using antisense oligonucleotide against survivin in an orthotopic transplant model of human hepatocellular carcinoma in nude mice. Oligonucleotides 2006; 16: 365–374.

    Article  CAS  Google Scholar 

  11. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC . Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998; 391: 806–811.

    Article  CAS  Google Scholar 

  12. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T . Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 2001; 411: 494–498.

    Article  CAS  Google Scholar 

  13. Stevenson M . Therapeutic potential of RNA interference. N Engl J Med 2004; 351: 1772–1777.

    Article  Google Scholar 

  14. Tang ZY . Hepatocellular carcinoma. J Gastroenterol Hepatol 2000; 15 (Suppl): G1–G7.

    Article  Google Scholar 

  15. Ito T, Shiraki K, Sugimoto K, Yamanaka T, Fujikawa K, Ito M et al. Survivin promotes cell proliferation in human hepatocellular carcinoma. Hepatology 2000; 31: 1080–1085.

    Article  CAS  Google Scholar 

  16. Suzuki A, Ito T, Kawano H, Hayashida M, Hayasaki Y, Tsutomi Y et al. Survivin initiates procaspase 3/p21 complex formation as a result of interaction with Cdk4 to resist Fas-mediated cell death. Oncogene 2000; 19: 1346–1353.

    Article  CAS  Google Scholar 

  17. Wu L, Wang Y, Tian D . Knockdown of survivin expression by siRNA induces apoptosis of hepatocellular carcinoma cells. J Huazhong Univ Sci Technolog Med Sci 2007; 27: 403–406.

    Article  CAS  Google Scholar 

  18. He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B . A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA 1998; 95: 2509–2514.

    Article  CAS  Google Scholar 

  19. Elbashir SM, Lendeckel W, Tuschl T . RNA interference is mediated by 21- and 22-nucleotide RNAs. Genes Dev 2001; 15: 188–200.

    Article  CAS  Google Scholar 

  20. Li K, Lin SY, Brunicardi FC, Seu P . Use of RNA interference to target cyclin E-overexpressing hepatocellular carcinoma. Cancer Res 2003; 63: 3593–3597.

    CAS  PubMed  Google Scholar 

  21. Li H, Fu X, Chen Y, Hong Y, Tan Y, Cao H et al. Use of adenovirusdelivered siRNA to target oncoprotein p28GANK in hepatocellular carcinoma. Gastroenterology 2005; 128: 2029–2041.

    Article  CAS  Google Scholar 

  22. Cho-Rok J, Yoo J, Jang YJ, Kim S, Chu IS, Yeom YI et al. Adenovirus-mediated transfer of siRNA against PTTG1 inhibits liver cancer cell growth in vitro and in vivo. Hepatology 2006; 43: 1042–1052.

    Article  Google Scholar 

  23. Altieri DC . Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer 2008; 8: 61–70.

    Article  CAS  Google Scholar 

  24. Liu J, Du W, Fan D . Survivin, the promising target in hepatocellular carcinoma gene therapy. Cancer Biol Ther 2008; 7: 555–556.

    Article  CAS  Google Scholar 

  25. Zhu DE, Höti N, Song Z, Jin L, Wu Z, Wu Q et al. Suppression of tumor growth using a recombinant adenoviral vector carrying the dominant-negative mutant gene Survivin-D53A in a nude mice model. Cancer Gene Ther 2006; 13: 762–770.

    Article  CAS  Google Scholar 

  26. Weihua Z, Tsan R, Huang WC, Wu Q, Chiu CH, Fidler IJ et al. Survival of cancer cells is maintained by EGFR independent of its kinase activity. Cancer Cell 2008; 13: 385–393.

    Article  Google Scholar 

  27. Bertrand JR, Pottier M, Vekris A . Comparison of antisense oligonucleotides and siRNAs in cell culture and in vivo. Biochem Biophys Res Commun 2002; 296: 1000–1004.

    Article  CAS  Google Scholar 

  28. Kumar R, Conklin DS, Mittal V . High-Throughput Selection of Effective RNAi Probes for Gene silencing. Genome Res 2003; 13: 2333–2340.

    Article  CAS  Google Scholar 

  29. Tamm I, Wang Y, Sausville E, Scudiero DA, Vigna N, Oltersdorf T et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs. Cancer Res 1998; 58: 5315–5320.

    CAS  Google Scholar 

  30. Strand S, Hofmann WJ, Hug H, Müller M, Otto G, Strand D et al. Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand expressing tumor cells—a mechanism of immune evasion? Nat Med 1996; 2: 1361–1366.

    Article  CAS  Google Scholar 

  31. Suzuki A, Tsutomi Y, Akahane K, Araki T, Miura M . Resistance to Fas-mediated apoptosis: activation of caspase 3 is regulated by cell cycle regulator p21WAF1 and IAP gene family ILP. Oncogene 1998; 17: 931–939.

    Article  CAS  Google Scholar 

  32. Sugimoto K, Shiraki K, Ito T, Fujikawa K, Takase K, Tameda Y et al. Expression of functional CD40 in human hepatocellular carcinoma. Hepatology 1999; 30: 920–926.

    Article  CAS  Google Scholar 

  33. Fei Q, Zhang H, Fu L, Dai X, Gao B, Ni M et al. Experimental cancer gene therapy by multiple anti-survivin hammerhead ribozymes. Acta Biochim Biophys Sin 2008; 40: 466–477.

    Article  CAS  Google Scholar 

  34. Yamaguchi Y, Shiraki K, Fuke H, Inoue T, Miyashita K, Yamanaka Y et al. Targeting X-linked inhibitor of apoptosis protein or survivin by short interfering RNAs sensitize hepatoma cells to TNF-related apoptosis-inducing ligand- and chemotherapeutic agent-induced cell death. Oncol Rep 2005; 14: 1311–1316.

    CAS  PubMed  Google Scholar 

  35. He SQ, Rehman H, Gong MG, Zhao YZ, Huang ZY, Li CH et al. Inhibiting survivin expression enhances TRAIL-induced tumoricidal activity in human hepatocellular carcinoma via cell cycle arrest. Cancer Biol Ther 2007; 46: 1247–1257.

    Google Scholar 

  36. Peng Z . Current status of gendicine in China: recombinant human Ad-p53 agent for treatment of cancers. Hum Gene Ther 2005; 16: 1016–1027.

    Article  CAS  Google Scholar 

  37. Dummer R, Rochlitz C, Velu T, Acres B, Limacher JM, Bleuzen P et al. Intralesional adenovirus-mediated interleukin-2 gene transfer for advanced solid cancers and melanoma. Mol Ther 2008; 16: 985–994.

    Article  CAS  Google Scholar 

  38. Shen C, Buck AK, Liu X, Winkler M, Reske SN . Gene silencing by adenovirus-delivered siRNA. FEBS Lett 2003; 539: 111–114.

    Article  CAS  Google Scholar 

  39. Verma IM, Somia N . Gene therapy—promises, problems and prospects. Nature 1997; 389: 239–242.

    Article  CAS  Google Scholar 

  40. Vile R . Cancer gene therapy—new approaches to tumour cell killing. J Gene Med 2000; 2: 141–143.

    Article  CAS  Google Scholar 

  41. Li Q, Kay MA, Finegold M, Stratford-Perricaudet LD, Woo SL . Assessment of recombinant adenoviral vectors for hepatic gene therapy. Hum Gene Ther 1993; 4: 403–409.

    Article  CAS  Google Scholar 

  42. Kay MA, Graham F, Leland F, Woo SL . Therapeutic serum concentration of human alpha-1-antitrypsin after adenoviral mediated gene transfer into mouse hepatocytes. Hepatology 1995; 21: 815–819.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Basic Research Program of China (No. 2004CB518805), the National Natural Scientific Foundation of China (No.30873006), the Program for Changjiang Scholars and Innovative Research Team in University (No. IRT0459).

Author information

Author notes

  1. R Zhang and L Ma: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Shannxi Province, Xi’an, China

    R Zhang, L Ma, J Zhao, L Jia, L Yao & A Yang

  2. Department of Hepatobiliary Surgery, Xijing Hospital, Fourth Military Medical University, Shannxi Province, Xi’an, China

    L Ma & K Li

  3. Department of Genetics and Development, Fourth Military Medical University, Shannxi Province, Xi’an, China

    M Zheng & H Han

  4. Department of Immunology, Fourth Military Medical University, Shannxi Province, Xi’an, China

    J Ren, T Wang, Y Meng & A Yang

Authors
  1. R Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. T Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. L Jia
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. L Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. H Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. K Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. A Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A Yang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, R., Ma, L., Zheng, M. et al. Survivin knockdown by short hairpin RNA abrogates the growth of human hepatocellular carcinoma xenografts in nude mice. Cancer Gene Ther 17, 275–288 (2010). https://doi.org/10.1038/cgt.2009.68

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/cgt.2009.68

Keywords

This article is cited by

Search

Advanced search

Quick links