Visualization of cancer cells requires distinguishing malignant from normal cells by objective criteria with high specificity. For several years, tumor markers expressed on the surface of cancer cells have been characterized as cancer signatures, and their labeling with specific imaging probes has revolutionized cancer diagnosis. This specific labeling is also an important tool in surgery tumor ablation. The present study considers the tumor labeling potential of an aptamer that specifically recognizes the epithelial cancer biomarker mucin1 (MUC1). This anti-MUC1 aptamer was investigated in vitro in a three-dimensional (3D) environment and compared to an anti-MUC1 antibody for its capacity to visualize cancer cells. Multicellular spheroids of breast cancer MCF-7 cells were used as tumor models and anti-MUC1 fluorescent aptamer and antibody were visualized by fluorescence imaging. Results showed that the antibodies interacted only with cells located on the surface of the spheroid, whereas the anti-MUC1 aptamers were able to penetrate inside these 3D tumor models and thereafter internalized into the cancer cells. Due to their lack of immunogenicity and their facility to be chemically modified, aptamers may replace advantageously the use of antibodies in diagnosis based on imaging setup thanks to their specific detection of cancer cells without invasive surgical procedures or during clinical intraoperative intervention.
Get full access to this article
View all access options for this article.
References
1.
BAOUENDIM., COGNETJ.A., FERREIRAC.S., MISSAILIDISS., COUTANTJ., PIOTTOM., HANTZE., and HERVE DU PENHOATC. (2012). Solution structure of a truncated anti-MUC1 DNA aptamer determined by mesoscale modeling and NMR. FEBS J., 279, 479–490.
CAIL., CHENZ.Z., CHENM.Y., TANGH.W., and PANGD.W. (2013). MUC-1 aptamer-conjugated dye-doped silica nanoparticles for MCF-7 cells detection. Biomaterials, 34, 371–381.
4.
CERCHIAL., and DE FRANCISCISV. (2010). Targeting cancer cells with nucleic acid aptamers. Trends Biotechnol., 28, 517–525.
5.
CHANGM., YANGC.S., and HUANGD.M. (2011). Aptamer-conjugated DNA icosahedral nanoparticles as a carrier of doxorubicin for cancer therapy. ACS Nano., 5, 6156–6163.
6.
CHARLTONJ., SENNELLOJ., and SMITHD. (1997). In vivo imaging of inflammation using an aptamer inhibitor of human neutrophil elastase. Chem. Biol., 4, 809–816.
7.
CHENL.Q., XIAOS.J., HUP.P., PENGL., MAJ., LUOL.F., LIY.F., and HUANGC.Z. (2012). Aptamer-mediated nanoparticle-based protein labeling platform for intracellular imaging and tracking endocytosis dynamics. Anal. Chem., 84, 3099–3110.
8.
CHUT.C., MARKSJ.W.3RD, LAVERYL.A., FAULKNERS., ROSENBLUMM.G., ELLINGTONA.D., and LEVYM. (2006). Aptamer:toxin conjugates that specifically target prostate tumor cells. Cancer Res., 66, 5989–5992.
9.
CIBIELA., PESTOURIEC., and DUCONGEF. (2012). In vivo uses of aptamers selected against cell surface biomarkers for therapy and molecular imaging. Biochimie, 94, 1595–1606.
10.
COHENS., and MARGELS. (2012). Engineering of near IR fluorescent albumin nanoparticles for in vivo detection of colon cancer. J. Nanobiotechnology, 10, 36.
11.
DA PIEVEC., PERKINSA.C., and MISSAILIDISS. (2009). Anti-MUC1 aptamers: radiolabelling with Tc-99m and biodistribution in MCF-7 tumour-bearing mice. Nucl. Med. Biol., 36, 703–710.
12.
DAIF., ZHANGY., ZHUX., SHANN., and CHENY. (2012). Anticancer role of MUC1 aptamer-miR-29b chimera in epithelial ovarian carcinoma cells through regulation of PTEN methylation. Target Oncol., 7, 217–725.
13.
DASSIEJ.P., LIUX.Y., THOMASG.S., WHITAKERR.M., THIELK.W., STOCKDALEK.R., MEYERHOLZD.K., MCCAFFREYA.P., MCNAMARAJ.O.2ND, and GIANGRANDEP.H. (2009). Systemic administration of optimized aptamer-siRNA chimeras promotes regression of PSMA-expressing tumors. Nat. Biotechnol., 27, 839–849.
14.
FAROKHZADO.C., CHENGJ., TEPLYB.A., SHERIFII., JONS., KANTOFFP.W., RICHIEJ.P., and LANGERR. (2006). Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. Proc. Natl. Acad. Sci. U. S. A., 103, 6315–6320.
15.
FERREIRAC.S., MATTHEWSC.S., and MISSAILIDISS. (2006). DNA aptamers that bind to MUC1 tumour marker: design and characterization of MUC1-binding single-stranded DNA aptamers. Tumour Biol., 27, 289–301.
16.
GAOX., CUIY., LEVENSONR.M., CHUNGL.W., and NIES. (2004). In vivo cancer targeting and imaging with semiconductor quantum dots. Nat. Biotechnol., 22, 969–976.
17.
GIBOTL., and ROLSM.P. (2013). Progress and prospects: the use of 3D spheroid model as a relevant way to study and optimize DNA electrotransfer. Curr. Gene Ther., 13, 175–181.
18.
GIBOTL., WASUNGUL., TEISSIEJ., and ROLSM.P. (2013). Antitumor drug delivery in multicellular spheroids by electropermeabilization. J. Control. Release, 167, 138–147.
19.
GRAFFC.P., and WITTRUPK.D. (2003). Theoretical analysis of antibody targeting of tumor spheroids: importance of dosage for penetration, and affinity for retention. Cancer Res., 63, 1288–1296.
20.
HOFFMANR.M. (1993). To do tissue culture in two or three dimensions? That is the question. Stem Cells, 11, 105–111.
21.
HORMT.M., and SCHROEDERJ.A. (2013). MUC1 and metastatic cancer: Expression, function and therapeutic targeting. Cell Adh. Migr., 7, 187–198.
22.
HUY., DUANJ., ZHANQ., WANGF., LUX., and YANGX.-D. (2012). Novel MUC1 aptamer selectively delivers cytotoxic agent to cancer cells in vitro. Plos One, 7, e31970.
23.
JINE., ZHANGB., SUNX., ZHOUZ., MAX., SUNQ., TANGJ., SHENY., VAN KIRKE., MURDOCHW.J., et al. (2013). Acid-active cell-penetrating peptides for in vivo tumor-targeted drug delivery. J. Am. Chem. Soc., 135, 933–940.
24.
KANWARJ.R., ROYK., and KANWARR.K. (2011). Chimeric aptamers in cancer cell-targeted drug delivery. Crit. Rev. Biochem. Mol. Biol., 46, 459–477.
25.
KAURS., VENKTARAMANG., JAINM., SENAPATIS., GARGP.K., and BATRAS.K. (2012). Recent trends in antibody-based oncologic imaging. Cancer Lett., 315, 97–111.
26.
KAUSHALS., MCELROYM.K., LUIKENG.A., TALAMINIM.A., MOOSSAA.R., HOFFMANR.M., and BOUVETM. (2008). Fluorophore-conjugated anti-CEA antibody for the intraoperative imaging of pancreatic and colorectal cancer. J. Gastrointest. Surg., 12, 1938–1950.
27.
KIMJ.K., CHOIK.J., LEEM., JOM.H., and KIMS. (2012). Molecular imaging of a cancer-targeting theragnostics probe using a nucleolin aptamer- and microRNA-221 molecular beacon-conjugated nanoparticle. Biomaterials, 33, 207–217.
28.
KUROSAKIT., HIGUCHIN., KAWAKAMIS., HIGUCHIY., NAKAMURAT., KITAHARAT., HASHIDAM., and SASAKIH. (2012). Self-assemble gene delivery system for molecular targeting using nucleic acid aptamer. Gene, 491, 205–209.
29.
LIX., ZHAOQ., and QIUL. (2013). Smart ligand: aptamer-mediated targeted delivery of chemotherapeutic drugs and siRNA for cancer therapy. J. Control. Release, 171, 152–162.
30.
LIUN., ZHOUC., ZHAOJ., and CHENY. (2012). Reversal of paclitaxel resistance in epithelial ovarian carcinoma cells by a MUC1 aptamer-let-7i chimera. Cancer Invest., 30, 577–582.
31.
MAEDAH. (2001). The enhanced permeability and retention (EPR) effect in tumor vasculature: the key role of tumor-selective macromolecular drug targeting. Adv. Enzyme Regul., 41, 189–207.
32.
MAYERG. (2009). The chemical biology of aptamers. Angew Chem. Int. Ed. Engl., 48, 2672–2689.
33.
MCELROYM., KAUSHALS., LUIKENG.A., TALAMINIM.A., MOOSSAA.R., HOFFMANR.M., and BOUVETM. (2008). Imaging of primary and metastatic pancreatic cancer using a fluorophore-conjugated anti-CA19-9 antibody for surgical navigation. World J. Surg., 32, 1057–1066.
34.
MCNAMARAJ.O.2ND, ANDRECHEKE.R., WANGY., VILESK.D., REMPELR.E., GILBOAE., SULLENGERB.A., and GIANGRANDEP.H. (2006). Cell type-specific delivery of siRNAs with aptamer-siRNA chimeras. Nat. Biotechnol., 24, 1005–1015.
35.
ORAVAE.W., CICMILN., and GARIEPYJ. (2010). Delivering cargoes into cancer cells using DNA aptamers targeting internalized surface portals. Biochim. Biophys. Acta, 1798, 2190–2200.
36.
RUIGROKV.J., LEVISSONM., EPPINKM.H., SMIDTH., and VAN DER OOSTJ. (2011). Alternative affinity tools: more attractive than antibodies?. Biochem. J., 436, 1–13.
37.
SANDVIGK., LLORENTEA., RODALS.K., EKERP., GARREDO., STAHLHUTM., VAN DEURSB. (2000). Apical macropinocytosis in polarized MDCK cells: regulation by N-ethylmaleimide-sensitive proteins. Eur. J. Cell Biol., 79, 447–457.
38.
SHIH., HEX.X., WANGK.M., WUX., YEX.S., GUOQ.P., TANW.H., QINGZ.H., YANGX.H., and ZHOUB. (2011). Activatable aptamer probe for contrast-enhanced in vivo cancer imaging based on cell membrane protein-triggered conformation alteration. Proc. Natl. Acad. Sci. U. S. A., 108, 3900–3905.
39.
SHIGDARS., QIAOL., ZHOUS.F., XIANGD., WANGT., LIY., LIML.Y., KONGL., LIL., and DUANW. (2013). RNA aptamers targeting cancer stem cell marker CD133. Cancer Lett., 330, 84–95.
40.
SUTHERLANDR., BUCHEGGERF., SCHREYERM., VACCAA., and MACHJ.P. (1987). Penetration and binding of radiolabeled anti-carcinoembryonic antigen monoclonal antibodies and their antigen binding fragments in human colon multicellular tumor spheroids. Cancer Res., 47, 1627–1633.
41.
TERWISSCHA VAN SCHELTINGAA.G., VAN DAMG.M., NAGENGASTW.B., NTZIACHRISTOSV., HOLLEMAH., HEREKJ.L., SCHRODERC.P., KOSTERINKJ.G., LUB-DE HOOGM.N., and DE VRIESE.G. (2011). Intraoperative near-infrared fluorescence tumor imaging with vascular endothelial growth factor and human epidermal growth factor receptor 2 targeting antibodies. J. Nucl. Med., 52, 1778–1785.
42.
THURBERG.M., and WITTRUPK.D. (2008). Quantitative spatiotemporal analysis of antibody fragment diffusion and endocytic consumption in tumor spheroids. Cancer Res., 68, 3334–3341.
43.
THURBERG.M., ZAJICS.C., and WITTRUPK.D. (2007). Theoretic criteria for antibody penetration into solid tumors and micrometastases. J. Nucl. Med., 48, 995–999.
44.
WANGJ., and LIG. (2011). Aptamers against cell surface receptors: selection, modification and application. Curr. Med. Chem., 18, 4107–4116.
45.
WENNINGL.A., and MURPHYR.M. (1999). Coupled cellular trafficking and diffusional limitations in delivery of immunotoxins to multicell tumor spheroids. Biotechnol. Bioeng., 62, 562–575.
46.
WORNA., and PLUCKTHUNA. (2001). Stability engineering of antibody single-chain Fv fragments. J. Mol. Biol., 305, 989–1010.
47.
WUP., GAOY., ZHANGH., and CAIC. (2012). Aptamer-guided silver-gold bimetallic nanostructures with highly active surface-enhanced Raman scattering for specific detection and near-infrared photothermal therapy of human breast cancer cells. Anal. Chem., 84, 7692–7699.
48.
YUC., HUY., DUANJ., YUANW., WANGC., XUH., and YANGX.-D. (2011). Novel aptamer-nanoparticle bioconjugates enhances delivery of anticancer drug to MUC1-positive cancer cells in vitro. Plos One, 6, e24077.
49.
ZHUG.Z., YEM., DONOVANM.J., SONGE.Q., ZHAOZ.L., and TANW.H. (2012). Nucleic acid aptamers: an emerging frontier in cancer therapy. Chem. Commun., 48, 10472–10480.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
