Skip to main content

Advertisement

SpringerLink
Log in

siRNA Directed Against Survivin Enhances Pancreatic Cancer Cell Gemcitabine Chemosensitivity

  • Original Article
  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Survivin is known to be overexpressed in various human malignancies, including pancreatic cancer, and to cause resistance to radiation and chemotherapy, so the regulation of this molecule could be a new strategy for treating pancreatic cancer. In our study, a short interfering RNA (siRNA) plasmid expression vector against survivin was constructed and transfected into human pancreatic cancer cell lines of Panc-1 and BxPC3. The expression of survivin mRNA and protein among the stable transfected cells and the untransfected cells was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. Tumor cell growth in vitro was assessed by trypan blue exclusion. The cell cycle distribution and cell apoptosis were measured by flow cytometry. The cytotoxicity assay was measured by the MTT test. Our results showed that survivin siRNA treatment caused a specific and profound decrease of survivin mRNA and protein that was associated with decreased cell growth, spontaneous apoptosis, and a specific G0/G1 arrest. Furthermore, the suppression of survivin can enhance the chemosensitivity of pancreatic cancer cells to gemcitabine significantly. We suggest that the RNAi against survivin gene strategy would be a potential approach to chemosensitization therapy in human pancreatic cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Keighley MRB (2003) Gastrointestinal cancers in Europe. Aliment Pharmacol Ther 18:7–30. doi:10.1046/j.0953-0673.2003.01722.x

    Article  PubMed  Google Scholar 

  2. Pino SM, Xiong HQ, McConkey D, Abbruzzese JL (2004) Novel therapies for pancreatic adenocarcinoma. Curr Gastroenterol Rep 6:119–125. doi:10.1007/s11894-004-0038-x

    Article  PubMed  Google Scholar 

  3. El-Rayes BF, Philip PA (2003) A review of systemic therapy for advanced pancreatic cancer. Clin Adv Hematol Oncol 1:430–434

    PubMed  Google Scholar 

  4. Wheatley SP, McNeish IA (2005) Survivin: a protein with dual roles in mitosis and apoptosis. Int Rev Cytol 247:35–88. doi:10.1016/S0074-7696(05)47002-3

    Article  PubMed  CAS  Google Scholar 

  5. Lee MA, Park GS, Lee HJ, Jung JH, Kang JH, Hong YS, Lee KS, Kim DG, Kim SN (2005) Survivin expression and its clinical significance in pancreatic cancer. BMC Cancer 5:127. doi:10.1186/1471-2407-5-127

    Article  PubMed  Google Scholar 

  6. Kami K, Doi R, Koizumi M, Toyoda E, Mori T, Ito D, Fujimoto K, Wada M, Miyatake S, Imamura M (2004) Survivin expression is a prognostic marker in pancreatic cancer patients. Surgery 136:443–448. doi:10.1016/j.surg.2004.05.023

    Article  PubMed  Google Scholar 

  7. Satoh K, Kaneko K, Hirota M, Masamune A, Satoh A, Shimosegawa T (2001) Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors. Cancer 92:271–278. doi: 10.1002/1097-0142(20010715)92:2<271::AID-CNCR1319>3.0.CO;2-0

    Article  PubMed  CAS  Google Scholar 

  8. Wang Y, Zhu H, Quan L, Zhou C, Bai J, Zhang G, Zhan Q, Xu N (2005) Downregulation of survivin by RNAi inhibits the growth of esophageal carcinoma cells. Cancer Biol Ther 4:974–978

    Article  PubMed  CAS  Google Scholar 

  9. Kami K, Doi R, Koizumi M, Toyoda E, Mori T, Ito D, Kawaguchi Y, Fujimoto K, Wada M, Miyatake S, Imamura M (2005) Downregulation of survivin by siRNA diminishes radioresistance of pancreatic cancer cells. Surgery 138:299–305. doi:10.1016/j.surg.2005.05.009

    Article  PubMed  Google Scholar 

  10. Song H, Xin XY, Xiao F, Wang DT, Yue QH, Han X (2008) Survivin gene RNA interference inhibits proliferation, induces apoptosis, and enhances radiosensitivity in HeLa cells. Eur J Obstet Gynecol Reprod Biol 136:83–89. doi:10.1016/j.ejogrb.2006.07.057

    Article  PubMed  CAS  Google Scholar 

  11. Liu F, Xie ZH, Cai GP, Jiang YY (2007) The effect of survivin on multidrug resistance mediated by P-glycoprotein in MCF-7 and its adriamycin resistant cells. Biol Pharm Bull 30:2279–2283. doi:10.1248/bpb.30.2279

    Article  PubMed  CAS  Google Scholar 

  12. Fuessel S, Herrmann J, Ning S, Kotzsch M, Kraemer K, Schmidt U, Hakenberg OW, Wirth MP, Meye A (2006) Chemosensitization of bladder cancer cells by survivin-directed antisense oligodeoxynucleotides and siRNA. Cancer Lett 232:243–254. doi:10.1016/j.canlet.2005.02.027

    Article  PubMed  CAS  Google Scholar 

  13. Tran J, Master Z, Yu JL, Rak J, Dumont DJ, Kerbel RS (2002) A role for survivin in chemoresistance of endothelial cells mediated by VEGF. Proc Natl Acad Sci USA 99:4349–4354. doi:10.1073/pnas.072586399

    Article  PubMed  CAS  Google Scholar 

  14. Li H, Niederkorn JY, Neelam S, Alizadeh H (2006) Downregulation of survivin expression enhances sensitivity of cultured uveal melanoma cells to cisplatin treatment. Exp Eye Res 83:176–182. doi:10.1016/j.exer.2005.11.024

    Article  PubMed  CAS  Google Scholar 

  15. Shinohara ET, Hallahan DE, Lu B (2004) The use of antisense oligonucleotides in evaluating survivin as a therapeutic target for radiation sensitization in lung cancer. Biol Proced Online 6:250–256. doi:10.1251/bpo95

    Article  PubMed  CAS  Google Scholar 

  16. Asanuma K, Kobayashi D, Furuya D, Tsuji N, Yagihashi A, Watanabe N (2002) A role for survivin in radioresistance of pancreatic cancer cells. Jpn J Cancer Res 93:1057–1062

    PubMed  CAS  Google Scholar 

  17. Pennati M, Binda M, Colella G, Folini M, Citti L, Villa R, Daidone MG, Zaffaroni N (2003) Radiosensitization of human melanoma cells by ribozyme-mediated inhibition of survivin expression. J Invest Dermatol 120:648–654. doi:10.1046/j.1523-1747.2003.12082.x

    Article  PubMed  CAS  Google Scholar 

  18. Zhu K, Qin H, Cha SC, Neelapu SS, Overwijk W, Lizee GA, Abbruzzese JL, Hwu P, Radvanyi L, Kwak LW, Chang DZ (2007) Survivin DNA vaccine generated specific antitumor effects in pancreatic carcinoma and lymphoma mouse models. Vaccine 25:7955–7961

    Article  PubMed  CAS  Google Scholar 

  19. Hannon GJ (2002) RNA interference. Nature 418:244–251. doi:10.1038/418244a

    Article  PubMed  CAS  Google Scholar 

  20. Marx J (2000) Interfering with gene expression. Science 288:1370–1372. doi:10.1126/science.288.5470.1370

    Article  PubMed  CAS  Google Scholar 

  21. Aoki Y, Cioca DP, Oidaira H, Kamiya J, Kiyosawa K (2003) RNA interference may be more potent than antisense RNA in human cancer cell lines. Clin Exp Pharmacol Physiol 30:96–102. doi:10.1046/j.1440-1681.2003.03801.x

    Article  PubMed  CAS  Google Scholar 

  22. Brantl S (2002) Antisense-RNA regulation and RNA interference. Biochim Biophys Acta 1575:15–25

    PubMed  CAS  Google Scholar 

  23. Matta H, Hozayev B, Tomar R, Chugh P, Chaudhary PM (2003) Use of lentiviral vectors for delivery of small interfering RNA. Cancer Biol Ther 2:206–210

    PubMed  CAS  Google Scholar 

  24. Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553. doi:10.1126/science.1068999

    Article  PubMed  CAS  Google Scholar 

  25. Lens SM, Wolthuis RM, Klompmaker R, Kauw J, Agami R, Brummelkamp T, Kops G, Medema RH (2003) Survivin is required for a sustained spindle checkpoint arrest in response to lack of tension. EMBO J 22:2934–2947. doi:10.1093/emboj/cdg307

    Article  PubMed  CAS  Google Scholar 

  26. Lens SM, Vader G, Medema RH (2006) The case for Survivin as mitotic regulator. Curr Opin Cell Biol 18:616–622. doi:10.1016/j.ceb.2006.08.016

    Article  PubMed  CAS  Google Scholar 

  27. Shin S, Sung BJ, Cho YS, Kim HJ, Ha NC, Hwang JI, Chung CW, Jung YK, Oh BH (2001) An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7. Biochemistry 40:1117–1123. doi:10.1021/bi001603q

    Article  PubMed  CAS  Google Scholar 

  28. Altieri DC (2001) The molecular basis and potential role of survivin in cancer diagnosis and therapy. Trends Mol Med 7:542–547. doi:10.1016/S1471-4914(01)02243-2

    Article  PubMed  CAS  Google Scholar 

  29. Chandele A, Prasad V, Jagtap JC, Shukla R, Shastry PR (2004) Upregulation of survivin in G2/M cells and inhibition of caspase 9 activity enhances resistance in staurosporine-induced apoptosis. Neoplasia 6:29–40

    PubMed  CAS  Google Scholar 

  30. Song Z, Yao X, Wu M (2003) Direct interaction between survivin and Smac/DIABLO is essential for the anti-apoptotic activity of survivin during taxol-induced apoptosis. J Biol Chem 278:23130–23140. doi:10.1074/jbc.M300957200

    Article  PubMed  CAS  Google Scholar 

  31. Li F, Ambrosini G, Chu EY, Plescia J, Tognin S, Marchisio PC, Altieri DC (1998) Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 396:580–584. doi:10.1038/25141

    Article  PubMed  CAS  Google Scholar 

  32. Takizawa BT, Uchio EM, Cohen JJ, Wheeler MA, Weiss RM (2007) Downregulation of survivin is associated with reductions in TNF receptors’ mRNA and protein and alterations in nuclear factor kappa B signaling in urothelial cancer cells. Cancer Invest 25:678–684. doi:10.1080/07357900701600954

    Article  PubMed  CAS  Google Scholar 

  33. Gazzaniga P, Silvestri I, Gradilone A, Scarpa S, Morrone S, Gandini O, Gianni W, Frati L, Aglianò AM (2007) Gemcitabine-induced apoptosis in 5637 cell line: an in-vitro model for high-risk superficial bladder cancer. Anticancer Drugs 18:179–185. doi:10.1097/CAD.0b013e328010ef47

    Article  PubMed  CAS  Google Scholar 

  34. Chang Q, Liu ZR, Wang DY, Kumar M, Chen YB, Qin RY (2004) Survivin expression induced by doxorubicin in cholangiocarcinoma. World J Gastroenterol 10:415–418

    PubMed  CAS  Google Scholar 

  35. Zhang B, Pan JS, Liu JY, Han SP, Hu G, Wang B (2006) Effects of chemotherapy and/or radiotherapy on survivin expression in ovarian cancer. Methods Find Exp Clin Pharmacol 28:619–625. doi:10.1358/mf.2006.28.9.1003581

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Jin Huang (Institute of Liver Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P.R. China) for the technical assistance. This work was supported by the National Natural Science Foundation of China, grant nos. 30471693 and 30600592.

Author information

Authors and Affiliations

  1. Department of Biliary-Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People’s Republic of China

    Wen-Song Liu, Ren-Yi Qin, Rui Tian, Min Wang, Jian-Xin Jiang, Ming Shen & Cheng-Jian Shi

  2. State Key Laboratory of Tumors, Cancer Center, Sun Yat-sen University, Guangzhou, 510060, People’s Republic of China

    Hai-Jiao Yan

Authors
  1. Wen-Song Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hai-Jiao Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ren-Yi Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rui Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jian-Xin Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ming Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Cheng-Jian Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ren-Yi Qin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, WS., Yan, HJ., Qin, RY. et al. siRNA Directed Against Survivin Enhances Pancreatic Cancer Cell Gemcitabine Chemosensitivity. Dig Dis Sci 54, 89–96 (2009). https://doi.org/10.1007/s10620-008-0329-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10620-008-0329-4

Keywords

Search

Navigation