Skip to main content
SpringerLink
Log in

Effects of combined siRNA-TR and -TERT on telomerase activity and growth of bladder transitional cell cancer BIU-87 cells

  • Published:
Journal of Huazhong University of Science and Technology [Medical Sciences] Aims and scope Submit manuscript

Summary

The effects of combined RNA interference (RNAi) of human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) genes on telomerase activity in a bladder cancer cell line (BIU-87 cells) were investigated by using gene chip technology in vitro with an attempt to evaluate the role of RNAi in the gene therapy of bladder transitional cell cancer (BTCC). Three TR-specific double-stranded small interfering RNAs (siRNAs) and three TERT-specific double-stranded siRNAs were designed to target different regions of TR and TERT mRNA. The phTR-siRNA, phTERT-siRNA, and the combination of both plasmids phTR+phTERT-siRNA were transfected into BIU-87 cells. The expression of hTR and hTERT mRNA was detected by quantitative fluorescent reverse transcription-polymerase chain reaction, and a telomeric repeat amplification protocol was applied to detect telomerase activity. Growth inhibition of BIU-87 cells was measured by MTT assay. Gene chip analysis was performed to evaluate the effects of the combined RNAi of hTR+hTERT genes on telomerase activity and growth of BIU-87 cells in vitro. The results showed that the expression of hTERT and hTR mRNA was inhibited by pRNAT-hTERT-III, pRNAT-hTR-III, and pRNAT-hTR-III+hTERT-III in BIU-87 cells. The inhibition efficiency of pRNAT-hTERT-III, pRNAT-hTR-III, pRNAT-hTERT-III+pRNAT-hTR-III was 67% for TERT mRNA, 41% for TR mRNA, 57% for TR mRNA and 70% for TERT mRNA in BIU-87 cells respectively. The growth of BIU-87 cells was inhibited and telomerase activity was considerably decreased, especially in the cells treated with combined RNAi-hTR and -hTERT. Gene chip analysis revealed that 21 genes were down-regulated (ATM, BAX, BCL2, BCL2L1, BIRC5, CD44, CTNNB1, E2F1, JUN, MCAM, MTA1, MYC, NFKB1, NFKBIA, NME4, PNN, PNN, SERPINE1, THBS1, TNFRSF1A, and UCC1). The results indicated that hTR-siRNA and hTERT-siRNA, especially their combination, siRNA hTR+hTERT, specifically and effectively suppressed the expression of both hTR and hTERT mRNA and telomerase activity. Molecular biological mechanism by which combined siRNA-TR and -TERT inhibited telomerase activity and growth of BIU-87 cells in vitro may involve the down-regulation of the 21 genes.

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

Similar content being viewed by others

References

  1. Nakatsukasa H, Onishi T, Kaneyoshi T, et al. Expression of telomerase-associated protein 1 and telomerase reverse transcriptase in hepatocellular carcinoma. Br J Cancer 2000,82(4):833–837

    Article  PubMed  Google Scholar 

  2. Lord RV, Salonga D, Danenberg KD, et al. Telomerase reverse transcriptase expression is increased early in the Barrett’s metaplasia, dysplasia, adenocarcinoma sequence. J Gastrointest Surg, 2000,4(2):135–142

    Article  CAS  PubMed  Google Scholar 

  3. Konagi Y, Kabayashi D, Yajima T, et al. Telomerase activity is down regulated via decreases in hTERT but not TEP1 mRNA or hTERC during the differentiation of leukemic cells. Anticancer Res, 2000,20(2A):773–778

    Google Scholar 

  4. Watanabe N. Telomerase, cell immortality and cancer Hokkaido Igaku Zasshi, 2001, 76(3):127–132

    CAS  Google Scholar 

  5. Rhya MS. Telomeres telomerase and immortality. J Natl Cancer Ins, 1995,87(12):884–894

    Article  Google Scholar 

  6. Zou L, Zhang P, Luo C, et al. shRNA targeted hTERT suppress cell proliferation of bladder cancer by inhibiting telomerase activity. Cancer Chemother Pharmacol, 2006, 57(3):328–334

    Article  CAS  PubMed  Google Scholar 

  7. Ito H, Kyo S, Kanaya T, et al. Expression of human telomerase subunit and correlation with telomerase activity in urothelial cancer. Clin Cancer Res, 1998,4(7):1603–1608

    CAS  PubMed  Google Scholar 

  8. Lancelin F, Anidjar M, Villette JM, et al. Telomerase activity as a potential marker in preneoplastic bladder lesions. BJU Int, 2000,85(4):526–531

    Article  CAS  PubMed  Google Scholar 

  9. Suzuki T, Suzuki Y, Fujioka T. Expression of the catalytic subunit associated with telomerase gene in human urinary bladder cancer. J Urol, 1999,162(6):2217–2220

    Article  CAS  PubMed  Google Scholar 

  10. Glasgow JN, Everts M, Curiel DT. Transductional targeting of adenovirus vectors for gene therapy. Cancer Gene Ther, 2006,13(9):830–844

    Article  CAS  PubMed  Google Scholar 

  11. Elbashir SM, Lendeckel W, Tuschl T. RNA interference is mediated by 21- and 22- nucleotide RNAs. Gene Dev, 2001,15(2):188–200

    Article  CAS  PubMed  Google Scholar 

  12. Oeggerli M, Tomovska S, Schraml P, et al. E2F3 amplification and overexpression is associated with invasive tumor growth and rapid tumor cell proliferation in urinary bladder cancer. Oncogene, 2004, 23(33):5616–5623

    Article  CAS  PubMed  Google Scholar 

  13. Wu X, Deng Y, Wang G, Tao K. Combining siRNAs at two different sites in the EGFR to suppress its expression, induce apoptosis, and enhance 5-fluorouracil sensitivity of colon cancer cells. J Surg Res, 2007,183(1):56–63

    Article  Google Scholar 

  14. Lin Y, Uemura H, Fujinami K, et al. Telomerase activity in primary prostate cancer. J Urol, 1997,157(3):1161–1165

    Article  CAS  PubMed  Google Scholar 

  15. Wu KJ, Grandori C, Amacker M, et al. Direct activation of Tert transcription by c-myc. Nature Genet, 1999,21:220–224 (2)

    Article  CAS  PubMed  Google Scholar 

  16. Suenaga M, Yamaguchi A, Soda H, et al. Antiproliferative effects of Gefitinib are associated with suppression of E2F1 expression and telomerase activity. Anticancer research, 2006,26(5A):3387–3392

    CAS  PubMed  Google Scholar 

  17. Pirocanac EC, Nassirpour R, Yang M, et al. Bax-induction gene therapy of pancreatic cancer. J Surg Res, 2002, 106(2):346–351

    Article  CAS  PubMed  Google Scholar 

  18. Malerba I, Gribaldo L, Diodovich C. Induction of apoptosis and inhibition of telomerase activity in human bone marrow and HL-60 p53 null cells treated with anti-cancer drugs. Toxicology in Vitro, 2005,19(4):523–532

    Article  CAS  PubMed  Google Scholar 

  19. Sandra Angèle, Alison Falconer, et al. ATM protein overexpression in prostate tumors. Anatomic Pathology, 2004,121(2):231–236

    Google Scholar 

  20. David Feldser, Margaret A. Strong, Carol W. Greider, et al. Ataxia telangiectasia mutated (Atm) is not required for telomerase-mediated elongation of short telomerase. Cancer Res, 2003(63):8188–8196

    Google Scholar 

  21. Shiloh Y. ATM and related protein kinases: safeguarding genome integrity. Nat Rev Cancer, 2003,3(3):155–168

    Article  CAS  PubMed  Google Scholar 

  22. Metcalfe JA, Parkhill J, Campbell L, et al. Accelerated telomere shortening in ataxia telangiectasia. Nat-Genet, 1996,13(3):350–353

    Article  CAS  PubMed  Google Scholar 

  23. Greenwell PW, Kronmoal SL, Porter SE. et al. TEL1, a gene involved in controlling telomere length in S. cerevisiae, is homologous to the human ataxia telangiectasia gene. Cell, 1995,82(5):823–829

    Article  CAS  PubMed  Google Scholar 

  24. DuBois ML, Haimberger ZW, Mclntosh MW, et al. A quantitative assay for telomere protection in Saccharomyces cerevisiae. Genetics, 2002,161(3):995–1013

    CAS  PubMed  Google Scholar 

  25. Fujimoto K, Takahashi M. Telomerase activity in human leukemic cell lines is inhibited by antisense pentadecadeoxynucleotides targeted against C-myc mRNA. Biochem Biophys Res Commun, 2007,24(3):775–781

    Google Scholar 

  26. Latil A, Vidaud D, Valeri A, et al. hTERT expression correlates with Myc over-expression in human prostate cancer. Int J Cancer, 2000,89(2):172–1797

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Urology, Jinling Hospital, Nanjing, 210002, China

    Wen Cheng  (程 文), Zhifeng Wei  (位志峰), Jianping Gao  (高建平), Zhengyu Zhang  (张征宇), Jingping Ge  (葛京平), Kangzhen Jing  (景抗震), Feng Xu  (徐 锋) & Peng Xie  (解 鹏)

Authors
  1. Wen Cheng  (程 文)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhifeng Wei  (位志峰)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianping Gao  (高建平)
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhengyu Zhang  (张征宇)
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jingping Ge  (葛京平)
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kangzhen Jing  (景抗震)
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Feng Xu  (徐 锋)
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Peng Xie  (解 鹏)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jianping Gao  (高建平).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cheng, W., Wei, Z., Gao, J. et al. Effects of combined siRNA-TR and -TERT on telomerase activity and growth of bladder transitional cell cancer BIU-87 cells. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 30, 391–396 (2010). https://doi.org/10.1007/s11596-010-0363-2

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11596-010-0363-2

Key words

Search

Navigation