全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
PLOS ONE  2012 

Use of Cell-SELEX to Generate DNA Aptamers as Molecular Probes of HPV-Associated Cervical Cancer Cells

DOI: 10.1371/journal.pone.0036103

Full-Text   Cite this paper   Add to My Lib

Abstract:

Background Disease-specific biomarkers are an important tool for the timely and effective management of pathological conditions, including determination of susceptibility, diagnosis, and monitoring efficacy of preventive or therapeutic strategies. Aptamers, comprising single-stranded or double-stranded DNA or RNA, can serve as biomarkers of disease or biological states. Aptamers can bind to specific epitopes on macromolecules by virtue of their three dimensional structures and, much like antibodies, aptamers can be used to target specific epitopes on the basis of their molecular shape. The Systematic Evolution of Ligands by EXponential enrichment (SELEX) is the approach used to select high affinity aptamers for specific macromolecular targets from among the >1013 oligomers comprising typical random oligomer libraries. In the present study, we used live cell-based SELEX to identify DNA aptamers which recognize cell surface differences between HPV-transformed cervical carcinoma cancer cells and isogenic, nontumorigenic, revertant cell lines. Methodology/Principal Findings Whole-cell SELEX methodology was adapted for use with adherent cell lines (which we termed Adherent Cell-SELEX (AC-SELEX)). Using this approach, we identified high affinity aptamers (nanomolar range Kd) to epitopes specific to the cell surface of two nontumorigenic, nontumorigenic revertants derived from the human cervical cancer HeLa cell line, and demonstrated the loss of these epitopes in another human papillomavirus transformed cervical cancer cell line (SiHa). We also performed preliminary investigation of the aptamer epitopes and their binding characteristics. Conclusions/Significance Using AC-SELEX we have generated several aptamers that have high affinity and specificity to the nontumorigenic, revertant of HPV-transformed cervical cancer cells. These aptamers can be used to identify new biomarkers that are related to carcinogenesis. Panels of aptamers, such as these may be useful in predicting the tumorigenic potential and properties of cancer biopsies and aid in the effective management of pathological conditions (diagnosis, predicted outcome, and treatment options).

References

[1]  Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, et al. (1999) Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. The Journal of pathology 189: 12–19.
[2]  Centers for Disease Control and Prevention: Division of Cancer Prevention and Control (2010) Human Papillomavirus (HPV)-Associated Cancers.
[3]  Liu X, Clements A, Zhao K, Marmorstein R (2006) Structure of the human Papillomavirus E7 oncoprotein and its mechanism for inactivation of the retinoblastoma tumor suppressor. J Biol Chem 281: 578–586.
[4]  Chaturvedi AK, Engels EA, Pfeiffer RM, Hernandez BY, Xiao W, et al. Human papillomavirus and rising oropharyngeal cancer incidence in the United States. J Clin Oncol 29: 4294–4301.
[5]  Kyrgiou M, Shafi M (2008) HPV Vaccine. Obstet Gynaecol Reprod Med 19: 26–28.
[6]  Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249: 505–510.
[7]  Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346: 818–822.
[8]  Shangguan D, Li Y, Tang Z, Cao ZC, Chen HW, et al. (2006) Aptamers evolved from live cells as effective molecular probes for cancer study. Proc Natl Acad Sci U S A 103: 11838–11843.
[9]  Shangguan D, Cao ZC, Li Y, Tan W (2007) Aptamers evolved from cultured cancer cells reveal molecular differences of cancer cells in patient samples. Clin Chem 53: 1153–1155.
[10]  Athanassiou M, Hu Y, Jing L, Houle B, Zarbl H, et al. (1999) Stabilization and reactivation of the p53 tumor suppressor protein in nontumorigenic revertants of HeLa cervical cancer cells. Cell Growth Differ 10: 729–737.
[11]  Boylan MO, Athanassiou M, Houle B, Wang Y, Zarbl H (1996) Activation of tumor suppressor genes in nontumorigenic revertants of the HeLa cervical carcinoma cell line. Cell Growth Differ 7: 725–735.
[12]  Summerhayes IC, Lampidis TJ, Bernal SD, Nadakavukaren JJ, Nadakavukaren KK, et al. (1982) Unusual retention of rhodamine 123 by mitochondria in muscle and carcinoma cells. Proc Natl Acad Sci U S A 79: 5292–5296.
[13]  Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31: 3406–3415.
[14]  Morris KN, Jensen KB, Julin CM, Weil M, Gold L (1998) High affinity ligands from in vitro selection: complex targets. Proc Natl Acad Sci U S A 95: 2902–2907.
[15]  Osborne SE, Ellington AD (1997) Nucleic Acid Selection and the Challenge of Combinatorial Chemistry. Chem Rev 97: 349–370.
[16]  Nutiu R, Li Y (2005) In vitro selection of structure-switching signaling aptamers. Angew Chem Int Ed Engl 44: 1061–1065.
[17]  Wilson DS, Szostak JW (1999) In vitro selection of functional nucleic acids. Annu Rev Biochem 68: 611–647.
[18]  Jayasena SD (1999) Aptamers: an emerging class of molecules that rival antibodies in diagnostics. Clin Chem 45: 1628–1650.
[19]  Tang J, Xie J, Shao N, Yan Y (2006) The DNA aptamers that specifically recognize ricin toxin are selected by two in vitro selection methods. Electrophoresis 27: 1303–1311.
[20]  Tang J, Yu T, Guo L, Xie J, Shao N, et al. (2007) In vitro selection of DNA aptamer against abrin toxin and aptamer-based abrin direct detection. Biosens Bioelectron 22: 2456–2463.
[21]  Lee JH, Canny MD, De Erkenez A, Krilleke D, Ng YS, et al. (2005) A therapeutic aptamer inhibits angiogenesis by specifically targeting the heparin binding domain of VEGF165. Proc Natl Acad Sci U S A 102: 18902–18907.
[22]  Hermann T, Patel DJ (2000) Adaptive recognition by nucleic acid aptamers. Science 287: 820–825.
[23]  Cerchia L, Duconge F, Pestourie C, Boulay J, Aissouni Y, et al. (2005) Neutralizing aptamers from whole-cell SELEX inhibit the RET receptor tyrosine kinase. PLoS Biol 3: e123.
[24]  Siddiqui MA, Keating GM (2005) Pegaptanib: in exudative age-related macular degeneration. Drugs 65: 1571–1577; discussion 1578–1579.
[25]  Cunningham ET Jr, Adamis AP, Altaweel M, Aiello LP, Bressler NM, et al. (2005) A phase II randomized double-masked trial of pegaptanib, an anti-vascular endothelial growth factor aptamer, for diabetic macular edema. Ophthalmology 112: 1747–1757.
[26]  Ng EW, Adamis AP (2005) Targeting angiogenesis, the underlying disorder in neovascular age-related macular degeneration. Can J Ophthalmol 40: 352–368.
[27]  Nimjee SM, Keys JR, Pitoc GA, Quick G, Rusconi CP, et al. (2006) A novel antidote-controlled anticoagulant reduces thrombin generation and inflammation and improves cardiac function in cardiopulmonary bypass surgery. Mol Ther 14: 408–415.
[28]  Cerchia L, Hamm J, Libri D, Tavitian B, de Franciscis V (2002) Nucleic acid aptamers in cancer medicine. FEBS Lett 528: 12–16.
[29]  Cerchia L, de Franciscis V (2010) Targeting cancer cells with nucleic acid aptamers. Trends Biotechnol 28: 517–525.
[30]  Phillips JA, Lopez-Colon D, Zhu Z, Xu Y, Tan W (2008) Applications of aptamers in cancer cell biology. Anal Chim Acta 621: 101–108.
[31]  Watanabe M, Sheriff S, Lewis KB, Tinch SL, Cho J, et al. (2011) HMGA-targeted phosphorothioate DNA aptamers increase sensitivity to gemcitabine chemotherapy in human pancreatic cancer cell lines. Cancer Lett.
[32]  Orava EW, Cicmil N, Gariepy J (2010) Delivering cargoes into cancer cells using DNA aptamers targeting internalized surface portals. Biochim Biophys Acta 1798: 2190–2200.
[33]  Toscano-Garibay JD, Benitez-Hess ML, Alvarez-Salas LM (2011) Isolation and characterization of an RNA aptamer for the HPV-16 E7 oncoprotein. Arch Med Res 42: 88–96.
[34]  Laurenson S, Pett MR, Hoppe-Seyler K, Denk C, Hoppe-Seyler F, et al. Development of peptide aptamer microarrays for detection of HPV16 oncoproteins in cell extracts. Anal Biochem 410: 161–170.
[35]  Hirt B (1967) Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol 26: 365–369.
[36]  Musheev MU, Krylov SN (2006) Selection of aptamers by systematic evolution of ligands by exponential enrichment: addressing the polymerase chain reaction issue. Anal Chim Acta 564: 91–96.
[37]  Motulsky H, Christopoulos A (2004) Fitting Models to Biological Data using Linear and Nonlinear Regression. A Practical Guide to Curve Fitting. New York: Oxford University Press.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133