Zhang et al.

Journal of Hematology & Oncology (2019) 12:137

https://doi.org/10.1186/s13045-019-0833-3

REVIEW Open Access

Nanotechnology in cancer diagnosis:

progress, challenges and opportunities
Ye Zhang1†, Maoyu Li1,2†, Xiaomei Gao3, Yongheng Chen1* and Ting Liu2*

Abstract
In the fight against cancer, early detection is a key factor for successful treatment. However, the detection of cancer
in the early stage has been hindered by the intrinsic limits of conventional cancer diagnostic methods. Nanotechnology
provides high sensitivity, specificity, and multiplexed measurement capacity and has therefore been investigated for the
detection of extracellular cancer biomarkers and cancer cells, as well as for in vivo imaging. This review summarizes the
latest developments in nanotechnology applications for cancer diagnosis. In addition, the challenges in the translation of
nanotechnology-based diagnostic methods into clinical applications are discussed.
Keywords: Nanoparticle, Cancer diagnosis, Cancer biomarker

Background Moreover, cytology and histopathology cannot be effect-

Throughout the world, cancer mortality and incidence are ively and independently applied to detect cancer at an
increasing. As estimated by GLOBOCAN 2018, the number early stage [7]. Therefore, the development of technolo-
of new cancer cases will reach 18.1 million, and the number gies for detecting cancer at an early stage, before metas-
of cancer-related deaths will be 9.6 million [1, 2]. Predictions tasis, presents a major challenge.
suggest that by 2030, 30 million people will die from cancer Although nanotechnology has not yet been deployed clin-
each year [2]. In the fight against cancer, a key for successful ically for cancer diagnosis, it is already on the market in a
cancer treatment is early detection. Cancer-related mortality variety of medical tests and screens, such as the use of gold
is greatly reduced by early detection [3]. For example, breast nanoparticles in home pregnancy tests [8]. For cancer
cancer exhibits a 5-year relative survival rate of nearly 90% diagnosis, nanoparticles are being applied to capture cancer
at the local stage, while patients with distant metastasis biomarkers, such as cancer-associated proteins, circulating
exhibit a 5-year survival rate of only 27% [4]. tumor DNA, circulating tumor cells, and exosomes [9]. An
At present, imaging techniques and morphological essential advantage of applying nanoparticles for cancer de-
analysis of tissues (histopathology) or cells (cytology) aid tection lies in their large surface area to volume ratio relative
in early diagnosis of cancer. The most widely used im- to bulk materials [10]. Due to this property, nanoparticle
aging techniques, such as X-ray, magnetic resonance im- surfaces can be densely covered with antibodies, small
aging (MRI), computed tomography (CT), endoscopy, molecules, peptides, aptamers, and other moieties. These
and ultrasound, can only detect cancer when there is a moieties can bind and recognize specific cancer molecules
visible change to the tissue [5]. By that time, thousands (Fig. 1). By presenting various binding ligands to cancer
of cancer cells may have proliferated and even metasta- cells, multivalent effects can be achieved, which can improve
sized. In addition, current imaging methods cannot dis- the specificity and sensitivity of an assay [11].
tinguish benign lesions from malignant lesions [6]. Nanotechnology-based diagnostic methods are being
developed as promising tools for real-time, convenient,
and cost-effective cancer diagnosis and detection [12].
* Correspondence: yonghenc@163.com; liuting818@126.com This review summarizes recent progress in the develop-
†
Ye Zhang and Maoyu Li contributed equally to this work. ment of nanotechnology and addresses the application
1
Department of Oncology, NHC Key Laboratory of Cancer Proteomics, of nanotechnology in cancer diagnosis. We also provide
XiangYa Hospital, Central South University, Changsha 410008, China
2
Department of Gastroenterology, XiangYa Hospital, Central South University, our perspective on challenges in the use of nanotechnol-
Changsha 410008, China ogy for cancer diagnosis.
Full list of author information is available at the end of the article

© The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 2 of 13

Fig. 1 Nanotechnology improves cancer detection and diagnosis

Nanotechnology for the detection of extracellular cancer cancer). Specific interactions with antibodies, antibody frag-
biomarkers ments, or aptamers can help in the detection of these prop-
A cancer biomarker acts as a measurable biological mol- erties. The interaction event will then be converted into a
ecule that can be found in blood and other tissues or body quantifiable signal that can be measured [25].
fluids, such as saliva and urine, indicating that cancer exists In recent studies, QD-based biosensors have been used
in the body [13, 14]. Cancer biomarkers may be proteins for detecting cancer biomarkers. QDs are characterized by
(secreted proteins or cell surface proteins) [15], carbohy- a high quantum yield and molar extinction coefficient;
drates [16], or nucleic acids (circulating tumor DNA, wide absorption with narrow, high-efficiency Stokes shifts;
miRNA, etc.) [17] that are secreted by the body or cancer high resistance to photobleaching; and outstanding resist-
cells when cancer is present [18, 19]. The measurement of ance to degradation, which constitute unique properties
certain cancer biomarker levels enables early detection of [26, 27]. A sandwich-type assay is a common strategy for
cancer or tumor recurrence and helps monitor the efficacy detecting protein biomarkers and comprises many compo-
of the therapy. Nevertheless, the use of biomarkers has been nents, namely, a biomarker, a capture antibody, a second
limited by several barriers, including low biomarker con- capture antibody, and a secondary antibody that binds to
centrations in body fluids, heterogeneity in the abundance the capture antibody [7]. The secondary antibody can be
and timing of biomarkers within patients, and the difficulty visualized through various methods, such as staining and
in carrying out prospective studies [20]. Nanotechnology fluorescence [28].
offers high selectivity and sensitivity and the ability to con- In utilizing this strategy, two QD-conjugated antibodies
duct simultaneous measurements of multiple targets. Bio- against neuron-specific enolase (NSE) and carcinoembryo-
sensors can be improved with nanoparticles/nanomaterials nic antigen (CEA) were used to detect two biomarkers,
to provide specific targeting [21]. In addition, the use of and the limit of detection (LOD) of each reached 1.0 ng/
nanoparticles provides an increased surface-to-volume ml [29]. A zinc oxide (ZnO) QD-based sandwich immuno-
ratio, which makes biosensors more sensitive in fulfilling assay was developed for ZnO nanowire substrates, which
the demands of specific biomolecular diagnostics [22]. provided a large surface area that presents multiple bind-
Quantum dots (QDs), gold nanoparticles (AuNPs), and ing sites used for detection. CEA, the most popular cancer
polymer dots (PDs) are three common nanoparticle probes biomarker, has been applied for monitoring of anticancer
used in diagnosing cancer [23, 24]. treatment, as well as for prediction of tumor recurrence
following surgical resection in late-stage cancer patients,
Protein detection making it widely studied. NSE is an enzyme that catalyzes
A number of proteins have been granted FDA clearance for the conversion of 2-phosphoglycerate to phosphoenolpyr-
cancer detection, including CEA (colorectal cancer), AFP uvate, which shows a relationship with carcinoids, small
(liver cancer), PSA (prostate cancer), and CA-125 (ovarian cell lung carcinoma, and islet cell tumors. After secretion,
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 3 of 13

they could be detected at concentrations over 15 ng/mL, Similar to phospholipids, phosphate surfactants are
and the LOD of each reached 1.0 ng/mL. Another example beneficial for recognizing and cleaving sPLA-2 enzyme.
of an immunosensor based on QDs is ZnO QDs coated This unique characteristic could substantially help con-
with antibody against carbohydrate antigen 19-9 (CA 19-9, trol UCNP release, because the enzyme hydrolyzes an
a biomarker for pancreatic ductal adenocarcinoma), which ester group located between the fatty acid and ethylene
is an important application in the detection of CA 19-9. glycol, which instantly liberates the nanoparticles from
Electrostatic adsorption primarily aided in the the prostate cancer surface [38].
immobilization process on the basis of the high isoelectric
point of ZnO, and the immune reaction of the CA 19-9 ctDNA detection
antigens and antibodies produced the sandwich structure. Circulating tumor DNA (ctDNA) represents tumor-derived
CA 19-9 immunological recognition was converted into DNA fragments (approximately 100-200 base-pairs long) in
the detection of amplified signals presented by square the bloodstream [39]. ctDNA can be released from primary
wave stripping voltammetry (SWV), as well as the inherent tumors or circulating tumor cells (CTCs) and can allow
photoluminescence (PL) exhibited by the labeled QDs. detection of cancer through cancer-specific genetic aberra-
The dynamic range of the electrochemical assay was 0.1– tions. Detection of genetic aberrations in ctDNA can help
180 U/ml, and the LOD reached 0.04 U/ml, while the dy- detect cancer even before any sign of cancer occurs [40,
namic range exhibited by the optical spectral detection 41]. Highly specific hybridization with nucleic acid probes
was 1–180 U/ml, and the LOD reached 0.25 U/ml [30]. that have complementary sequences can be used to detect
Peptides are frequently applied to actively target cancer- cancer-associated genetic aberrations [35]. A DNA silver
ous tissues in vivo [31]. The Arg-Gly-Asp (RGD) peptide nanocluster (NC) fluorescent probe was developed for
motif is recognized by a receptor (integrin αvβ3) on the detection of a single exon in the BRCA1 gene in breast can-
cell surface implicated in cancer metastasis and angiogen- cer [42]. Under optimized conditions, this probe increased
esis and has been applied to target tumor tissue in vivo for the LOD to 6.4 × 10-11 M. Large deletion mutations in
diagnosis [32]. In one study, an iRGD (CRGDKGPDC)- BRCA1 were detected based on nanocluster fluorescence
mediated and enzyme-induced precise targeting gold upon hybridization induced by recognition. The certain
nanoparticle system (iRGD/AuNPs-A&C) was developed hybridization of the DNA-templated silver NC fluorescent
by simply co-administering the tumor-homing penetration probe to target DNAs was able to effectively enhance the
peptide iRGD with a legumain-responsive aggregable gold AgNC fluorescence, which had various intensities, thereby
nanoparticle [33]. iRGD/AuNPs-A&C showed high pene- distinguishing the BRCA1 deletion. When conditions were
tration and accumulation in 4 T1 mammary tumors [34]. optimal, there was an increase in the fluorescence intensity
Aptamers, which are single-stranded DNA (ssDNA) or presented by DNA-AgNCs at emission peaks near 440 nm
RNA sequences that can be isolated via exponential (excitation at 350 nm) as the presence of the deletion type
enrichment (SELEX) that relies on ligand systematic evolu- increased in a dynamic range of 1.0 × 10-10 to 2.4 × 10-6 M,
tion, can also be conjugated to nanoparticles [35, 36]. and the LOD reached 6.4 × 10-11 M. In this sensing system,
Aptamers have high affinity and strong binding specificity the deletion type led to higher fluorescence emission than
for their respective targets, such as ions, bacteria, peptides, the normal type, with the normal type typically showing
viruses, phospholipids, and even whole cells. A10 RNA low fluorescence.
aptamer-conjugated polymeric nanoparticles incorporating
Cy5 can bind to prostate-specific membrane antigen microRNA detection
(PSMA). Cy5-PLA/aptamer NPs could only bind to microRNA is associated with cancer diagnosis. Jou AF re-
LNCaP cells and canine prostate adenocarcinoma cells, ported a two-step sensing platform for sensitive detection
which are positive for PSMA but not to PC3 cells, which of miR-141, a promising biomarker for prostate cancer.
are negative for PSMA. Cy5-PLA NPs have been applied in The first step of the sensing platform used CdSe/ZnS QDs
balb/c mice and exhibit outstanding signals with low- modified with FRET quencher-functionalized nucleic
background fluorescence in different organs [37]. acids, which contained a telomerase primer sequence to-
Rare-earth upconverting nanophosphors (UCNPs) gether with a recognition sequence for the miR-141 recog-
promise to be a new generation of biological lumines- nition sequence. The FRET quencher exhibited covalent
cence labels. UCNPs are able to absorb radiation from binding to the nucleic acid-functionalized CdSe/ZnS QDs.
near-infrared (NIR) light and transform the radiation When miR-141 hybridized with the probe, a duplex was
into visual light by relying on the upconversion process formed, which would be cleaved by duplex-specific nucle-
after multiple-photon absorption. Overexpression of se- ase (DSN). The cleavage released the quencher unit and
creted phospholipase A2 (sPLA-2), an enzyme that cata- activated the fluorescence of the QDs. This cleavage also
lyzes phospholipid hydrolysis, has been reported to show led to exposure of the telomerase primer sequence. The
an association with prostate cancer cell proliferation. second step involved the primer unit elongation stimulated
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 4 of 13

by telomerase/dNTPs, incorporation of hemin, and chemi- invades the surrounding tissue and then enters the mi-
luminescence generated with the help of luminol/H2O2. crovasculature of the blood (intravasation) and lymph
This platform helped detect miR-141 in a serum sample systems, followed by survival and translocation through
and discriminated healthy individuals from prostate cancer the bloodstream to microvessels in distant tissues, subse-
carriers [43]. quent exit from the bloodstream (extravasation) and sur-
vival in the microenvironment of distant tissues, which
DNA methylation detection present a suitable foreign microenvironment for devel-
The genome methylation landscape (Methylscape) was re- opment of a macroscopic secondary tumor [48]. Early
cently reported as a common characteristic of most types detection of metastatic cancer cells in the bloodstream,
and cancers and therefore could be a common cancer bio- also known as circulating tumor cells (CTCs), can
marker [44]. In this study, the authors observed differ- potentially affect cancer prognosis and diagnosis.
ences between cancer genomes and normal genomes As a portion of a liquid biopsy, CTCs have been studied
based on DNA-gold affinity and DNA solvation and devel- broadly due to their potential applications. CTC detection
oped simple, quick, selective and sensitive electrochemical can help us understand the molecular organization of a
or colorimetric one-step assays to detect cancer. tumor in a minimally invasive manner. Nevertheless, CTCs
exhibit relatively low abundance and heterogeneity, which
Extracellular vesicle detection presents technical challenges for CTC isolation and
As circulating vesicles (30 nm–1 μm), extracellular vesicles characterization. In recent years, researchers have focused
(EVs) package molecular information, such as miRNA, on the application of nanotechnologies for sensitive detec-
DNA, protein, and mRNA, from mother cells and allow the tion of CTCs; these technologies can help characterize cells
detection of the molecular state of tumor cells that are diffi- and molecules, thereby enjoying broad clinical applications,
cult to access. In a recent study, the authors developed a such as disease detection at an early stage and evaluation of
new magnetic nanopore capture technique to isolate certain the treatment response and disease development.
subsets of extracellular vesicles (EVs) from plasma [45]. As demonstrated in many studies, it is possible for cell
The machine-learning and RNA-sequencing algorithms pseudopodia to form on surfaces with nanostructure,
helped identify EV miRNA biomarkers. This approach was thereby enhancing the local topographical interactions
applied to a mouse model of pancreatic ductal adenocarcin- between cancer cells and nanostructured substrates,
oma (PDAC) and contributed to identification of a bio- which is beneficial to CTC enrichment. For CTC detec-
marker panel of eleven EV miRNAs. tion, nanomaterials have an essential advantage in their
Recently, Tan et al. reported a sensor platform that pro- large surface-to-volume ratio, which enables adsorption
files proteins on the surface of exosomes within several mi- of high-efficiency targeting ligands with the ability to
nutes. The sensor consists of a gold nanoparticle (AuNP) recognize specific molecules on cancer cells; therefore,
and an aptamer panel under complexation, which was de- CTC isolation shows high specificity and recovery, and
signed based on 13-nm AuNPs that were noncovalently the detection sensitivity is enhanced.
conjugated with a panel of 5 aptamers that targeted cell People have reported different types of nanomaterials,
surface proteins with high affinity and strong specificity, as such as magnetic nanoparticles (MNPs), gold nanoparticles
demonstrated previously. The aptamers complexed with (AuNPs), quantum dots (QDs), nanowires, nanopillars, sili-
AuNPs prevented nanoparticle aggregation in a solution con nanopillars, carbon nanotubes, dendrimers, graphene
with high salt. Exosomes helped break the weak and non- oxide, and polymers, for CTC detection (Table 1) [61]. It
specific binding between the AuNP and aptamers, while has been demonstrated that these nanomaterials can im-
strong and specific binding between aptamers and exosome prove the sensitivity and specificity of CTC capture devices
surface proteins displaced aptamers from the AuNP sur- and have the potential to facilitate cancer diagnosis and
face, thereby facilitating AuNP aggregation. Due to aggrega- prognosis.
tion, the color of the AuNPs changed from red to blue, In the field of nanobiotechnology, MNPs are mature
indicating that the aptamers were bound to exosomal pro- nanomaterials that can bind to cells and have long been
teins. The intensity presented by the AuNP aggregation used for in vitro separation with the help of an external
(A650/A520) was indicative of the relative abundance ex- magnetic field [62]. Antibody-functionalized MNPs,
hibited by target proteins on the surface of exosomes [46]. namely, immunomagnetic nanoparticles, are frequently ap-
plied in the biomedical field. For CTC detection, immuno-
Nanotechnology for detection of cancer cells magnetic technologies usually specifically target EpCAM-
Detection of circulating tumor cells expressing CTCs with anti-EpCAM functionalized MNPs.
Approximately 90% of deaths from solid tumors are at- To perform single-cell transcriptional profiling of CTCs
tributed to metastasis [47]. In the course of metastatic purified from breast cancer patients, Powell et al .[63] used
dissemination, a cancer cell from the primary tumor first MagSweeper, which is an immunomagnetic enrichment
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 5 of 13

Table 1 Nanotechnology for detection of cancer cells

Nanoparticle Type of affinity probe Specificity ligand Type of cancer Reference
Magnetic nanoparticles Antibody EpCAM Colon/liver /lung/breast [49–51]
Quantum dots Aptamer PTK7 Leukemia [52]
Polymer dots Antibody EpCAM Breast [53]
Gold nanoparticles Aptamer Her2 Breast [54]
Antibody Cd2/cd3 Leukemia [55]
Carbon nanotubes Antibody EpCAM Liver [56]
Upconversion NPs Antibody Her2 Breast [57]
Nanorod arrays DNA aptamer EpCAM Breast [58]
Nanoparticle-coated silicon bead Antibody EpCAM/CD146 Breast [59]
Colorectal
Nanofibers Antibody EpCAM Breast [60]

device that can isolate tumor cells from unfractionated facilitated by hydrophobic interactions, during nanoparticle
blood. MagSweeper serves as a magnetic cell sorting system formation. A few amphiphilic polymers with functional
that uses magnetic rods covered by a sheath to sweep groups for subsequent covalent conjugation of biomole-
across capture wells and attract target cells labeled with cules, such as streptavidin and immunoglobulin G (IgG),
magnetic nanoparticles [64]. It can be used to acquire high- show co-condensation with most semiconducting poly-
purity CTCs from patient blood, while preserving their cap- mers for modification and functionalization of a nanoparti-
acity to initiate tumors and metastasize, facilitating robust cle surface. The PDs bioconjugates were able to label
analysis of single CTCs. Using the system, the authors suc- cellular targets in an effective and specific manner, includ-
cessfully purified CTCs from 70% of patients with primary ing a cell surface marker on human breast cancer cells,
and metastatic breast cancer and performed direct meas- with no need to detect nonspecific binding. The authors
urement of the gene expression in individual CTCs. demonstrated that the fluorescence exhibited by PD-
QDs are characterized by special optical properties, labeled MCF-7 cells was 25 times higher than that of QD-
which enhance their usefulness in cancer cell detection labeled cells and 18 times higher than that of Alexa
[65]. Due to their high quantum yields, QDs are helpful in Fluor-labeled cells, according to flow cytometry analysis
the detection of materials with low abundance. However, [67]. Based on the results, the authors believe that these
to enhance QD electrical characteristics, Pang et al. [52] ultrabright nanoparticles were successfully bioconju-
hybridized ZnO NDs and g-C3N4 QDs to afford higher gated. Thus, this strategy provides a new opportunity
photoelectron transfer and separation efficiency. Due to for applying versatile semiconducting polymers to dif-
the outstanding advantages, ZnO NDs and g-C3N4 QDs ferent fluorescence measurement methods in biomedi-
enjoyed the extended application, and a photocatalyzed cine and modern biology.
renewable self-powered cytosensing device was presented Upconversion nanoparticles (UCNPs) are usually selected
on the basis of ZnO NDs@g-C3N4 QDs. Through conju- for fluorescent labeling considering the ability to excite
gation to the membrane PTK7-specific aptamer Sgc8c, the UCNPs with near-infrared (NIR) light to infrared (IR) light
device was used to detect CCRF-CEM cells (human acute for generation of fluorescence emission in the visible region
lymphoblastic leukemia cells), which express PTK7. The of the spectrum, leading to minimal background noise. Fur-
results showed that the device offers better performance thermore, applying NIR light as the excitation source pre-
in terms of detection range, detection limit, selectivity, vents damage to normal tissues on one hand and allows
and reproducibility. It captured only CCRF-CEM cells deep tissue penetration on the other hand [68].
(500 cell/mL) and no other cell types, such as HL-60, Shen et al. [69] described a simple method to conjugate
K562, and HeLa cells. The authors think that the device multifunctional nanoparticles (MFNPs) assembled by the
could be an effective platform for monitoring the progres- formation of various layers with a monoclonal anti-HER2
sion of leukemia and shows great promise. antibody and confirmed that the MFNPs exhibited the spe-
Polymer nanoparticles derived from various conductive cific detection of breast cancer BT474 cells (biomarker
hydrophobic polymers have been applied to produce nano- HER2 positive) with a high signal-to-noise ratio. The
particles with high quantum yields, photostability and that MFNPs have an obvious core-shell structure of UCNP@-
are nontoxic. Therefore, PD is ideal for CTC detection. Fe3O4@Au coated with anti-HER2 antibody and poly(-
Wu et al. [66] reported a strategy for semiconducting poly- ethylene glycol) (PEG) and exhibited an outstanding
mer dots (PDs) functionalization through entrapment of dispersity in different aqueous solutions and a high signal-
heterogeneous polymer chains into a single dot, which was to-noise ratio. The authors revealed that the photothermal
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 6 of 13

effect exhibited a new high-localization feature at the EpCAM can be used as a cell surface biomarker. Hence,
single-cell level based under a continuous-wave near-IR anti-EpCAM molecules are often applied to screening of
laser. Using these nanoparticles, the authors exerted a CTCs. CTCs undergoing EMT would cause inefficient
photothermal effect at the single-cell level. positive sorting on the basis of EpCAM expression. There-
For differentiating types of cells and cancer states, the au- fore, another approach is to find supplemental or replace-
thors used AuNPs capped with ligands of different hydro- ment markers for EpCAM. Many cell surface markers,
phobicity and coated with green fluorescent protein (GFP). such as vimentin, androgen receptor, glycan, major vault
Because the capping ligands used showed different chemical protein (MVP), and fibroblast activation protein α (FAPα),
structures, each AuNP-GFP complex was related to cancer have been studied for the detection of CTCs. However, a
cells to a different degree, considering cell membrane majority of these markers are only specific to certain cells,
composition differences [70]. Magnetic biotargeting- and many markers do not exist after CTCs experienced
multifunctional nanobioprobes (MBMNs) were used to EMT. More mesenchymal CTCs can be seen in the meta-
detect and isolate a small subset of malignant cells from nor- static stages of cancer, and thus, seeking proper EMT
mal cells. CoFe2O4@BaTiO3 magnetoelectric nanoparticles markers to evaluate prognosis and metastasis in cancer pa-
distinguished different cancer cells from each other and tients is important. Here, we compile the recently identified
from their normal counterparts through a magnetoelectric cell surface protein markers for detection of CTCs in differ-
effect [71]. ent cancer types (Table 2).

Detection through cell surface protein recognition Detection based on mRNA

The main method to detect cancer cells relies on binding In addition to the detection of extracellular nucleic acids,
of nanoparticle probes conjugated with moieties (protein, nanoparticles have also been developed as intracellular
short peptides, antibodies, oligonucleotide aptamers) to nucleic acid sensors. Seferos et al. [85] demonstrated that
surface markers on cancer cells and on those entering it is possible to use novel gold nanoparticle probes modi-
cells and detecting genetic content. For the detection of fied by oligonucleotides hybridized to complements la-
cancer cells, such as CTCs, capture or isolation is the first beled with a fluorophore as transfection agents and
and most important stage. Although the cell physical cellular “nanoflares” to detect mRNA in living cells. Nano-
properties, such as size, deformability, and density, are flares overcome many challenges in the creation of effect-
sometimes used, capture is primarily based on the affinity ive and sensitive intracellular probes and show a large
of cell surface molecules on CTCs detected with materials signal-to-noise ratio and sensitivity to changes in the
such as antibodies or aptamers. Unique surface proteins number of RNA transcripts in cells. Nanoflares, which
on CTCs are the primary targets. show high orientation, dense oligonucleotide coating and
Since many studies have demonstrated that EpCAM is can enter cells without the need for cytotoxic transfection
highly expressed on CTCs from many human malignancies, agent s[86], are useful for detecting intracellular mRNA.

Table 2 Cell surface protein markers for CTC detection

Marker Type of cell Type of cancer Detection method Reference
EpCAM CTC Colorectal CellSearch [72]
Breast
Head and neck
EpCAM and FRα CTC Non-small cell lung cancer CellSearch [73]
Glycan CTC Breast Flow cytometry [74]
Vimentin CTC Gastrointestinal Confocal microscopy [75]
EMT CTC Prostate CellSearch [76]
CTC Sarcoma Flow cytometry [77]
Vimentin+PD-L1 CTC Colorectal, prostate cancer Confocal microscopy [78]
Synaptophysin CTC Castration resistant prostate cancer CellSearch [79]
Major vault protein Mesenchymal and Hepatocellular carcinomas Flow cytometry [80]
intermediate CTCs
Androgen receptor CTC Metastatic breast cancer Epic Sciences CTC platform [81]
p75 Neurotrophin receptor+EpCAM CTC Esophageal squamous cell carcinoma Flow cytometry [82]
Carbonic anhydrase 9 and CD147 CTC Clear cell renal cell carcinoma NanoVelcro [83]
Excision repair cross-complementation group 1 CTC Platinum resistance ovarian cancer Reverse-transcription PCR [84]
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 7 of 13

Meanwhile, researchers have developed nanoflares for sequence-specific manner, and as a result, the signal ex-
simultaneous intracellular detection of various mRNA tran- hibited a spectral shift due to dimer formation. They
scripts. In these multiplexed nanoflare studies, AuNPs demonstrated that their method is powerful and can
functionalized with 2-3 DNA recognition strands and later successfully detect, quantify, and differentiate between
hybridized with short complementary reporter strands were different BRCA1 splice variants with single-copy sensi-
generated as nanoflares. For example, the use of multi- tivity, thereby laying a foundation for quantitative,
plexed nanoflares to detect survivin in addition to actin has single-cell genetic profiling in the future.
been investigated for normalizing nanoflare fluorescence
differences in cellular uptake. Therefore, the technique is Nanotechnology for in vivo imaging
comparable with conventional qRT-PCR for quantification In addition to cancer diagnosis through ex vivo detec-
of intracellular mRNA but can be performed at the single tion of cancer cells and biomarkers in liquid biopsy sam-
live cell level. ples, identifying cancerous tissues in the body has many
In some cases, the nanoflare platform was expanded to advantages in diagnosing and treating cancer. A proper
quantify intracellular RNA and detect spatiotemporal nanoparticle probe for detecting cancer tissue should ex-
localization in living cells [87]. In this work, β-actin- hibit a long circulation time, be specific to tumor tissue
targeting nanoflares were incubated with HeLa cells and and present low toxicity to nearby healthy tissue [7].
presented an obviously different intracellular distribution, Current related studies have focused on nanoparticle
exhibiting strong colocalization with mitochondria, which probe accumulation in tumor tissue for diagnosing can-
has not been previously demonstrated. Further, Smart- cer in animal models, generally mouse models.
Flares were utilized for studying melanoma tumor cell Nanoparticle probes can preferentially accumulate in
heterogeneit y[88]. These Smart-Flares were able to quan- tumor tissues through active or positive targeting, thereby
tify genomic expression at the single-cell level, thus allowing imaging and diagnosis of cancer in vivo [91]. In-
expanding our knowledge of cancer and metastasis. Inves- teractions between nanoparticles and blood proteins, up-
tigating the heterogeneity of cancer cells is crucial for take and clearance by the reticuloendothelial system
identifying novel biomarkers for early cancer diagnosis. (RES), penetration into solid tumors, and optimized active
Halo et al. [89] reported nanoflares, which were applied (vs passive) targeting for diagnosis of cancer constitute the
to capture live circulating breast cancer cells. These nano- main clinical application barriers. Fortunately, many de-
flares could detect target mRNA in model metastatic breast velopments related to these aspects have been achieved.
cancer cell (MBC) lines in human blood and exhibited high
recovery and fidelity reaching 99%. They also used nano- Passive targeting
flares together with later cultured mammospheres to reim- By definition, passive targeting represents the preferen-
plant the retrieved live recurrent breast cancer cells into tial extravasation capacity of 10- to 150-nm nanoparti-
whole human blood. Only 100 live cancer cells could be cles from the bloodstream into tumor tissue. Because
detected per mL of blood. Relying on the NanoFlare the tight junctions between endothelial cells in new
technology, it was possible to simultaneously isolate and blood vessels in tumors do not form properly, nanoparti-
characterize intracellular live cancer cells from whole blood. cles can preferentially accumulate in tumor tissue [92].
The authors demonstrated the ability of nanoflares to This form of passive nanoparticle entry into the tumor
collect CTCs for future culture and study. In addition, microenvironment is called the enhanced permeability
nanoflares contribute to the technology of combining intra- and retention (EPR) effect, which was detected approxi-
cellular markers with cell-surface markers for dually identi- mately 30 years ago by studying macromolecule trans-
fying putative CTCs. The combined method is likely to port into tumor tissues [93].
enhance the function of more platforms to specifically iden- QDs are characterized by obvious photostability, tunable
tify CTCs and subpopulations of CTCs. The authors think emission, and high quantum yield, contributing to their
that nanoflares provide the first gene-based approach to de- wide application in tumor tissue imaging through passive
tect, isolate, and characterize live cancer cells in the blood accumulation dependent on the EPR effect. Hong et al. [94]
and are likely to contribute to cancer diagnosis, prognosis, reported the use of a new NIR-II fluorophore, six-armed
and prediction, as well as personalized treatment. PEG-Ag2S QDs, for imaging of subcutaneous xenograft
Lee et al. reported an approach based on an elegant 4T1 murine tumors. They monitored how the NIR-II signal
plasmonic nanoparticle network structure, generating a was distributed within the mice for a long period of time
plasmon-coupled dimer able to detect single mRNA var- (up to 24-h post-injection (p.i.)) and observed that the NIR-
iants [90]. They applied the method to the detection and II fluorescence of 6PEGAg2S QDs increased stably in the
quantification of BRCA1 mRNA splice variants in vitro tumor region and decreased in the skin and other organs in
and in vivo. Two probes conjugated to nanoparticles the range of 30 min p.i. to 24 h p.i. The in vivo QD
were connected to the BRCA1 mRNA target in a pharmacokinetics suggested extraordinary accumulation of
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 8 of 13

6PEG-Ag2S QDs in tumors (> 10% ID/gram, where % ID/ nanoparticle surface (opsonization), the in vivo trafficking,
gram denotes the concentration of the probe relative to the uptake, and clearance of nanoparticles are greatly chan-
injected dose (ID) percentage per gram of tissue) through ged. Using PEG to coat a nanoparticle surface reduces
the EPR effect. They assert that imaging with these NIR-II nonspecific adsorption of serum proteins and minimizes
QDs offered deep inner organ registration, dynamic tumor protein corona formation, which increases the circulating
contrast, and rapid tumor detection. time of the nanoparticle. PEGylation of various nanoparti-
Researchers have also applied AuNPs to in vivo tumor cles, such as AuNPs and QDs, results in a longer circula-
imaging through passive targeting. Lai et al. [95] reported tion time in the blood, as well as slow accumulation in the
that mercaptoundecanoic acid-coated AuNPs could iden- liver and spleen [100].
tify and track primary glioma cells at the inoculation sites It is expected that nanotechnology-based imaging can
in mouse brains. Furthermore, these particles detected improve the specificity and sensitivity of cancer diagnosis
tumor-associated microvasculature in detail. In some on the one hand and reduce toxicity on the other hand.
cases, chitosan nanoparticles have been used for in vivo Garrigue et al. [101] recently reported that harnessing
imaging through the EPR effect. Nam et al. [96] reported a nanoparticles and the “enhanced permeation and reten-
tumor-targeting nanoparticle for use as an underlying tion” (EPR) effect helped them develop an innovative
multimodal imaging probe through optical/MR (MR: mag- nanosystem for positron emission tomography (PET) im-
netic resonance) dual imaging based on self-assembled aging. The system adopts a self-assembling amphiphilic
glycol chitosan. Through chemical modification and dendrimer that retains various PET reporting units at ter-
conjugation, they developed stable chitosan nanoparticles minals. This dendrimer was able to self-assemble into
labeled with Cy5.5 and encapsulated by Gd(III) (Cy5.5- small uniform nanomicelles, which accumulated in tumors,
CNP-Gd(III)). The Cy5.5-CNP-Gd(III) were spherical, allowing effective PET imaging. Due to the dendrimeric
with a size of approximately 350 nm. According to cellular multivalence combined with the passive tumor targeting
experiments, Cy5.5-CNP-Gd(III) were taken up in an ef- mediated by EPR, the nanosystem exhibited better imaging
fective manner, and distribution in the cytoplasm was ob- sensitivity and stronger specificity, with PET signal ratios
served. After administration via the tail vein of tumor- that increased by approximately 14-fold in comparison
bearing mice, the nanoparticles localized in large numbers with the clinical gold standard 2-fluorodeoxyglucose ([18F]
in the tumor, which was detected via noninvasive NIR FDG). Moreover, the dendrimer displayed an outstanding
fluorescence together with an MR imaging system. The safety profile and good pharmacokinetics for PET imaging.
authors propose that the unique characteristics of the gly- The authors believe that their study contributes to the
col chitosan nanoparticles, such as blood stability, deform- development of dendrimer nanosystems for effective and
ability, and quick cellular uptake, may greatly affect their promising cancer imaging.
in vivo tumor targeting ability, which relied on the EPR ef-
fect. Their results revealed that Cy5.5-CNP-Gd(III) could Active targeting
potentially be applied as an optical/MR dual imaging agent In addition to tumor imaging with the help of nanoparti-
for detecting and treating cancer. cle accumulation via passive targeting based on the EPR
Nanoparticle size and shape affect the EPR effect. There- effect, scholars have implemented a large number of stud-
fore, these factors should be considered when designing ies on recognition of receptors on the cell surface for ac-
nanoparticle probes for high tumor accumulation. Nano- tive targeting of tumor tissues. Usually, these methods
particles with a size of less than 10 nm can be quickly increase the number of nanoparticles delivered to tumor
eliminated by the kidneys, minimizing their localization in tissue in each unit time, thereby enhancing the sensitivity
tumor tissue [97]. Anisotropic particles exhibit an en- exhibited by in vivo tumor detection methods [102]. For
hancement in circulation time, possibly because aniso- the detection of tumors at an early stage with high con-
tropic nanoparticles are less likely to permeate endothelial trast imaging, active tumor targeting achieves a better re-
gaps in the liver in the range of hundreds of nanometers sult than passive targeting that relies on the EPR effect.
to tens of micrometers. Silica-coated QDs of various thick- Levenson and Nie reported antibody-conjugated QDs to
nesses were used to explore the impact of nanoparticle size target PSMA for active tumor targeting. The in vivo im-
on tumor tissue accumulation [98]. The 12-nm QDs pene- aging results for 3 types of QD surface modifications were
trated the tumor tissue with minimal hindrance, while the examined: (1) COOH groups, (2) PEG groups, and (3)
60-nm QDs extravasated but remained in 10-μm blood PEG plus PSMA Ab (PEG-PSMA Ab). Consistent with
vessels. By contrast, the 120-nm QDs showed no appre- the histological examinations, the COOH probe did not
ciable extravasation. present any tumor signals, and only weak tumor signals
When nanoparticles contact a biological fluid, their sur- were observed with the PEG probe (passive targeting), but
face will be become covered with a “corona” of biological the PEG-PSMA Ab-conjugated probe (active targeting)
macromolecules [99]. As serum proteins adsorb onto a exhibited intense signals. The comparison confirmed the
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 9 of 13

conclusion of more efficient and much quicker active tar- changing the CG base pair number. One strand of DNA
geting of tumors with a tumor-specific ligand compared (called the capture strand), besides being a scaffold to
with passive targeting in terms of tumor permeation, re- carry the drug, can be used for capturing the gold NRs
tention, and uptake [103]. after being thiolated, and the complementary strand
A recent study showed the frequent application of (called the targeting strand) can be used for specifically
peptides to active targeting of cancerous tissues in vivo. targeting cells after being preconjugated with ligand.
The RGD peptide is recognized by a receptor (integrin Gold NRs are the model NIR light-to-heat transduc-
αvβ3) on the cell surface involved in cancer angiogenesis ers used for cancer thermotherapy and for denatur-
and metastasis and has been applied to the targeting of ation of double-stranded DNA under NIR irradiation,
tumor tissue in vivo for diagnosis [104]. In one study, an as a result, loaded drugs are released at the target site
iRGD-mediated and enzyme-induced precise targeting for chemotherapy [105].
gold nanoparticle system (iRGD/AuNPs-A&C) was de- For CEA-overexpressing solid tumors, AMG 211 is a
veloped by simply co-administering a tumor-homing potentially interesting new bispecific T-cell engager
penetration peptide iRGD with a legumain-responsive (BiTE) antibody construct. AMG 211 was labeled with
aggregable gold nanoparticle. Intravenously injected zirconium-89 (89Zr) or a fluorescent dye to evaluate its
compounds coupled to iRGD were bound to tumor ves- tumor-targeting properties. 89Zr-AMG211 microPET
sels and then spread to extravascular tumor parenchyma, imaging can be complemented with ex vivo biodistribu-
while traditional RGD peptides only transported cargo tion and tracer integrity analysis. 89Zr-AMG211 showed
into blood vessels. iRGD homes to tumors through three dose-dependent CEA-specific tumor targeting and
steps: the RGD motif shows a mediating effect on the localization in viable tumor tissue. It can be used to clin-
binding to αv integrins on the tumor endothelium, and ically evaluate the in vivo AMG 211 behavior [106]. For
then, a proteolytic cleavage imposes a binding motif for example, 89Zr-AMG211 demonstrates specific tumor up-
neuropilin-1, which regulates penetration into the cells. take in LS174T colorectal carcinoma xenografts, and
Conjugation to iRGD contributed to an obvious im- microPET images revealed tumor uptake of 89Zr-
provement in the sensitivity of the tumor imaging agents AMG211 up to 24 h after injection, whereas the nontu-
and the activity of the anti-tumor drug [34]. mor targeting BiTE antibody construct 89Zr-Mec14 did
Previous studies reported Gd(3+)-DOTA and RGD not show accumulation in LS174T xenografts.
(UCNP-Gd-RGD)-labeled upconversion nanoprobe for Oseledchyk et al. reported a surface-enhanced reson-
glioblastoma dual-modality imaging. To prepare UCNP- ance Raman scattering (SERRS) nanoparticle conjugated
Gd-RGD, the amine-functional upconversion nanoparticle with folate receptor for in vivo imaging of xenograft
core is PEGylated, followed by Gd(3+) DOTA conjugation SKOV-3 ovarian cancer cells transduced with green
and RGD labeling. The colloidal stability of the obtained fluorescent protein and luciferase in a mouse model.
UCNP-Gd-RGD is improved and the cytotoxicity reduced This method was termed Topically Applied Surface-
compared with the UCNP core counterpart. Additionally, Enhanced Resonance Raman Ratiometric Spectroscopy
the UCNP-Gd-RGD presented an intense upconversion (TAS3RS) and employed an effective ratiometric imaging
luminescence in the deep-red region and a 3-fold en- approach with nontargeted SERRS-NP (nt-NP) and anti-
hancement in T1 relaxivity compared with Gd(3+) DOTA. FR-SERRS-NP (αFR-NP) multiplexing, successfully de-
Considering the recognition between integrin αvβ3 recep- tecting tumor lesions in a murine model of human ovar-
tors and UCNP-Gd-RGD, the nanoprobe exhibited spe- ian adenocarcinoma despite the size or localization of
cific binding to U87MG cells under confocal microscopy the tumor. TAS3RS can be used to detect microscopic
and quantification of ICP-MS. Furthermore, according to residual tumors during surgery [107].
the UCNP-Gd-RGD in vivo upconversion fluorescence/
MR imaging experiments together with the ex vivo ana- Clinical trials of nanotechnology-based applications in
lysis, subcutaneous U87MG tumor xenografts presented cancer diagnosis
preferential retention [34]. Nanotechnology for cancer diagnosis or detection has
One study reported a platform based on DNA that can been extensively studied in the laboratory; however, clin-
be self-assembled into NIR-responsive NPs for cancer ical application is the ultimate destination of these studies.
treatment. The platform has 3 different functional compo- Currently, multiple nanotechnology-based cancer diagno-
nents: (1) complementary DNA strands, (2) gold nanorods sis approaches are in clinical trials (Table 3). For example,
(NRs) (50 nm × 10 nm), and (3) a polyethylene glycol researchers at MSKCC and Cornell University have devel-
(PEG) layer. The complementary DNA strands have oped silica-hybrid nanoparticles (C-dots) for PET imaging
sequential CG base pairs and offer some loading sites for of patients with metastatic melanoma or malignant brain
doxorubicin (Dox), which is a model chemotherapeutic tumors. These nanoparticles coupled with 124I-labeled cy-
drug. The drug loading could be precisely tuned by clo-[Arg-Gly-Asp-Tyr] (cRGDY) peptides that are able to
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 10 of 13

Table 3 List of clinical trials of nanotechnology-based applications in cancer diagnosis

Title Disease Nanotechnology Purpose Phase NCT number
Carbon Nanoparticles as lymph node tracer in Rectal cancer CNP (carbon Lymph node Not NCT03550001
rectal cancer after neoadjuvant radiochemotherapy nanoparticle) tracer applicable
Ferumoxytol-iron oxide nanoparticle magnetic Head and neck Iron oxide Imaging Early Phase NCT01895829
resonance dynamic contrast-enhanced MRI cancer nanoparticle 1
Application of carbon nanoparticles in laparoscopic Colorectal tumor CNP Tumor localization Not NCT03350945
colorectal surgery and lymph node applicable
mapping
Sentinel lymph node mapping in endometrial Endometrial neoplasms CNP Lymph node Not NCT03778255
cancer mapping applicable
Sentinel lymph node mapping in cervical cancer Uterine cervical CNP Lymph node Not NCT03778268
neoplasms mapping applicable
Targeted silica nanoparticles for real-time Head and neck Fluorescent cRGDY- Imaging/ Phase 1/ NCT02106598
image-guided intraoperative mapping of cancer Melanoma PEG-Cy5.5-C dots Localization Phase 2
nodal metastases Breast cancer
Colorectal cancer
Nanochip technology in monitoring treatment Lymphoma Immuno-tethered Monitoring/ Not NCT03656835
response and detecting relapse in participants lipoplex nanoparticle detection applicable
with diffuse large B-cell lymphoma
Diagnosis of gastric lesions with Na-nose Stomach diseases/ Nanosensors Diagnosis NCT01420588
Gastric cancer
PET imaging of patients with melanoma and Metastatic melanoma/ 124I-cRGDY-PEG- Localization Not NCT01266096
malignant brain tumors using a 124I-labeled malignant brain dots applicable
cRGDY silica nanomolecular particle tracer: a tumors
microdosing study
Imaging of patients with malignant brain Brain cancer 89Zr-DFO-cRGDY- PET imaging Phase 1 NCT03465618
tumors using 89Zr-cRGDY ultrasmall silica PEG-Cy5-C' dots
particle tracers: a phase 1 microdosing study

selectively bind to integrins can be used to probe be extensively investigated in large clinical sample pools
tumor cells. In addition, another group of researchers before NP-based assays can reach the clinical
(also from MSKCC) have developed fluorescent application.
cRGDY C-dots (cRGDY-PEG-Cy5.5-C dots) for lymph The second challenge lies in large-scale production of
node mapping, which can be used during surgery to nanoprobes that are highly sensitive, highly reprodu-
visualize lymph nodes with cancer. As the research cible, and have long-term storage stability at an accept-
progresses, it is predictable that more and more able cost [110]. The production of most of the current
nanotechnology-based cancer diagnostic methods will nanoprobes is performed under highly optimized con-
progress into the clinic. ditions in labs; however, it is still a big challenge to pro-
duce these probes in batches. Because the shape, size,
Challenges in clinical translation composition, charge, and surface coating of nanoprobes
Although there has been much promising progress in vary, the detection results also greatly vary. To
nanotechnology-based cancer diagnosis, only a few minimize batch-to-batch changes, the synthesis steps
examples have advanced to clinical trials [108]. To ac- and nanoprobe functionalization must be simplified. In
celerate the translation of nanotechnology into clinical addition, nanoprobes may tend to aggregate during
applications, many challenges need to be addressed. storage. Moreover, the cost-effectiveness of developing
The first challenge in nanotechnology-based cancer a nanotechnology-based platform must be taken into
diagnosis is reliability. To be applied in the clinic, it is consideration.
essential to obtain reliable and quantitative detection re- The third challenge is to develop NP-based devices
sults. Many factors can affect NP-based detection sig- with high sensitivity and that are easy to handle and
nals, including nonspecific binding of NP probes, cost-efficient. Most NP-based assays were prepared in
aggregation and unfit detection conditions [109]. Fluctu- academic laboratories, and many assays are unrealistic
ations in the signals can also be attributed to compli- for clinical translation. For example, complicated con-
cated body fluid compositions. From a clinical validation focal Raman microscopes were used to implement
perspective, assay reliability and reproducibility need to most studies based on SERS but are rarely present in
Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 11 of 13

hospitals or clinical laboratories. Successful develop- currently available cancer diagnostics in the clinic, a
ment of NP-based POC (point of care) devices will variety of NP-based assays showed improvement in
greatly facilitate clinical application of nanotechnology terms of selectivity and sensitivity or offered entirely
in cancer diagnosis. new capacities that could not be achieved with trad-
The fourth challenge is the possible toxicity of nano- itional approaches. These advances will improve the
particles induced by their systemic administration. This survival rate of cancer patients by enabling early de-
challenge is mostly related to NP-based imaging in vivo. tection. In addition, these advances could be used to
To apply novel nanoparticle probes to in vivo imaging, monitor cancer progress in response to treatment,
the possible toxicity of these nanoparticles should be which may contribute to the development of better
assessed. The properties of nanoparticles (such as shape, strategies for cancer treatment.
size, charge, surface chemistry, targeting ligands, and Over the last decade, great progress has been made in
composition) can influence their toxicity. In addition, the field of nanotechnology-based cancer diagnosis, and
the biodistribution, biodegradability, and pharmacoki- our understanding in this field has greatly improved. Al-
netic properties of nanoparticles should be considered. though only a few NP-based assays have advanced to clin-
ical trials, with close collaboration among researchers,
Conclusion engineers, and clinicians, nanotechnology-based cancer
The recent progress in nanotechnology-based applica- diagnosis is poised to move into the clinic in the near fu-
tion in cancer diagnosis has been summarized in this ture. With its high sensitivity, specificity, and multiplexed
review (Fig. 2). In the past 10 years, many efforts measurement capacity, nanotechnology provides great op-
have been made to develop assays for cancer diagno- portunities to improve cancer diagnosis, which will ultim-
sis based on nanotechnology. Compared with the ately lead to an improved cancer patient survival rate.

Fig. 2 Schematic illustration of nanotechnology applications in cancer diagnosis

Zhang et al. Journal of Hematology & Oncology (2019) 12:137 Page 12 of 13

Abbreviations 17. Ma, H.; Liu, J.; Ali, M. M.; Mahmood, M. A.; Labanieh, L.; Lu, M.; Iqbal, S. M.;
AuNPs: Gold nanoparticles: BiTE: Bispecific T-cell engagerBLI: Bioluminescence Zhang, Q.; Zhao, W.; Wan, Y., Chem Soc Rev, (2015) 44, 1240.
imaging: CA 19-9: Carbohydrate antigen 19-9CEA: Carcinoembryonic 18. Joseph, L., Circulating tumor cells and nucleic acids for tumor diagnosis,
antigenCT: Computed tomographyCTCs: Circulating tumor cellsctDNA: (2013).
Circulating tumor DNADox: DoxorubicinDSN: Duplex-specific nucleaseEPR: 19. Ponomaryova A, Rykova E, Cherdyntseva N, Morozkin E, Zaporozhchenko I,
Enhanced permeation and retentionEvs: Extracellular vesiclesMRI: Magnetic Skvortsova T, Dobrodeev A, Zav Yalov A, Tuzikov S, Vlassov V. Ejc
resonance imagingNC: NanoclusterNPs: NanoparticlesNSE: Neuron-specific Supplements. 2015;13:43.
enolasePDAC: Pancreatic ductal adenocarcinomaPDs: polymer dotsPEG: 20. Hull LC, Farrell D, Grodzinski P. Biotechnol Adv. 2014;32:666.
Polyethylene glycolPET: Positron emission tomographyPL: 21. Sharifi M, Avadi MR, Attar F, Dashtestani F, Ghorchian H, Rezayat SM,
photoluminescencePSMA: Prostate-specific membrane antigenQDs: Quantum Saboury AA, Falahati M. Biosens Bioelectron. 2018;126:773.
dotsSERRS: Surface-enhanced resonance Raman scatteringsPLA-2: Secreted 22. Doria, G.; Conde, J.; Veigas, B.; Giestas, L.; Almeida, C.; Assuncao, M.; Rosa, J.;
phospholipase A2ssDNA: Single-stranded DNASWV: Square wave stripping Baptista, P. V., Sensors (Basel), (2012) 12, 1657.
voltammetryTAS3RS: Topically Applied Surface-Enhanced Resonance Raman 23. Harun, N. A.; Benning, M. J.; Horrocks, B. R.; Fulton, D. A., Nanoscale, (2013) 5,
Ratiometric SpectroscopyUCNPs: Upconverting nanophosphors, 3817.
Upconversion nanoparticlesZnO: Zinc oxide 24. Zhang, H.; Lv, J.; Jia, Z., Sensors-Basel, (2017) 17, 1078.
25. Wu, T. L.; Sun, Y. C.; Chang, P. Y.; Tsao, K. C.; Sun, C. F.; Wu, J. T., J CLIN LAB
ANAL, (2010) 17, -.
Acknowledgements
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YC and TL designed the study and provided critical suggestions. YZ, MYL, 27. Freeman, R.; Willner, I., CHEM SOC REV, (2012) 41, 4067.
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Foundation of China (81372904, 81570537, 81974074 and 81902858); the 32. Yun, W.; Zhang, X.; Xiong, Z.; Zhen, C.; Fisher, D. R.; Shuang, L.; Gambhir, S. S.;
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1
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