Abstract
Objective
Development of novel approaches for quantitative analysis of gene expression in intact
tumour cells could provide new methods for the detection of cervical cancer. Molecular
beacons (MBs) are single-stranded oligonucleotide probes that form stem-and-loop structures.
Survivin, a member of the ‘inhibitor of apoptosis’ family of proteins, is highly expressed
in cervical cancer. The aim of this study was to determine the feasibility of MBs
targeting wild-type survivin in the diagnosis of cervical cancer.
Study design
MBs with oligonucleotide sequences complementing survivin mRNA and covalently linked
with FITC or Cy3 were designed and synthesized. The specificity and sensitivity of
survivin MBs were examined in human cervical cancer cell lines and smears from cervical
cancer patients, and confirmed by reverse transcriptase polymerase chain reaction
(RT-PCR), immunocytochemistry, Western blotting and the thinprep cytological test
(TCT).
Results
Both survivin MB-FITC and MB-Cy3 produced a strong fluorescent signal in cervical
cancer cells, and the intensity was consistent with the results from RT-PCR, Western
blotting and immunocytochemical staining. In the initial clinical cohort study, the
sensitivity of survivin MBs was 61.37% (27/44) and the specificity was 72.72% (34/44);
the sensitivities of Western blotting and TCT were 76.1% (32/42) and 68.19% (30/44),
respectively. No significant difference in sensitivity was observed between MBs, Western
blotting and TCT for different tumour grades and International Federation of Gynecology
and Obstetrics (FIGO) stages.
Conclusions
Survivin MBs are specific and sensitive molecular probes for the detection of cervical
cancer cells. They have great potential in the early detection and follow-up of patients
with cervical cancer.
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to European Journal of Obstetrics and Gynecology and Reproductive BiologyAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Global cancer statistics, 2002.CA Cancer J Clin. 2005; 55: 74-108
- Cervical cancer mortality in young women in Europe: patterns and trends.Eur J Cancer. 2000; 36: 2266-2271
- Cervical cancer: prevention, diagnosis, and therapeutics.CA Cancer J Clin. 2001; 51: 92-114
- Molecular beacons: probes that fluoresce upon hybridization.Nat Biotechnol. 1996; 14: 303-308
- Molecular beacons: a novel DNA probe for nucleic acid and protein studies.Chemistry. 2000; 6: 1107-1111
- Molecular beacons for detecting DNA binding proteins.Nat Biotechnol. 2002; 20: 171-176
- Stem–loop oligonucleotides: a robust tool for molecular biology and biotechnology.Trends Biotechnol. 2002; 20: 249-256
- IAP family proteins–suppressors of apoptosis.Genes Dev. 1999; 13: 239-252
- A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma.Nat Med. 1997; 3: 917-921
- The molecular basis and potential role of survivin in cancer diagnosis and therapy.Trends Mol Med. 2001; 7: 542-547
- HPV detection and measurement of HPV-16, telomerase and survivin transcripts in colposcopy clinic patients.J Clin Pathol. 2001; 54: 304-308
- Molecular beacon imaging of tumor marker gene expression in pancreatic cancer cells.Cancer Biol Ther. 2005; 4: 561-570
- Overexpression of PML induced apoptosis in bladder cancer cell by caspase dependent pathway.Cancer Lett. 2006; 236: 259-268
- Primary application study in early diagnosis of bladder cancer by survivin molecular beacons.Urology. 2007; 70: 60-64
- tRNA-linked molecular beacons for imaging mRNAs in the cytoplasm of living cells.Nucleic Acids Res. 2005; 33: 1902-1912
- Real-time detection of gene expression in cancer cells using molecular beacon imaging: new strategies for cancer research.Cancer Res. 2005; 65: 1909-1917
- Effects of human papillomavirus type 16 oncoproteins on survivin gene expression.J Gen Virol. 2006; 87: 87-294
- A novel strategy for human papillomavirus detection and genotyping with SybrGreen and molecular beacon polymerase chain reaction.Am J Pathol. 2001; 159: 1651-1660
- Current concepts in biomarker technology for bladder cancers.Clin Chem. 2000; 46: 595-605
- Preliminary study of diagnostic utility of molecular beacons in bladder cancer.Urology. 2010; 76: e8-e13
Article info
Publication history
Published online: July 04, 2011
Accepted:
June 15,
2011
Received in revised form:
April 27,
2011
Received:
November 23,
2010
Identification
Copyright
© 2011 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.