Precise Cancer Diagnosis Research Articles

SWCNTs@GQDs composites as nanocarriers for enzyme-free dual-signal amplification electrochemical immunoassay of cancer biomarker

Carcinoembryonic antigen (CEA) as a typical biomarker plays a remarkable role in various human physiological processes. A sensitive and rapid method should be developed to realize the precise diagnosis of cancer at its early stage. In this work, we successfully synthesized the peroxidase-like graphene quantum dots (GQDs) and utilized it to fabricate an enzyme-free electrochemical immunosensor towards CEA by forming a single-wall carbon nanotubes@GQDs (SWCNTs@GQDs) composite. In addition to the SWCNTs@GQDs, reduced graphene oxide-Au nanoparticles (rGO-Au NPs) with the huge specific surface area and excellent conductivity were employed to modify the electrodes, leading to a dual-signal amplification result. Both the electrochemical impedance spectroscopy and cyclic voltammetry measurements were performed to evaluate the interface properties of the layer-by-layer modified electrodes. The proposed electrochemical immunosensor displayed a good specificity and sensitivity towards the detection of CEA ranging from 50 pg mL−1 to 650 pg mL−1 with a low detection limit of 5.3 pg mL−1. This simple and inexpensive immunosensor shows promising potential for the determination of other biomarkers in clinical translation.

Fluorescent activatable gadofullerene nanoprobes as NIR-MR dual-modal in vivo imaging contrast agent.

Dual mode imaging technology is widely developed to achieve the early-stage precision cancer diagnosis. Here we designed a dual-modal magnetic resonance/near infrared fluorescence optical imaging contrast agent (GdF-SS-NIR783) with the fluorescence activatable and safer gadofullerene. The nanoprobes were fabricated by conjugating the gadofullerene derivatives with a NIR fluorescence imaging agent (NIR783) via the disulfide bond. The obtained nanoprobes showed no fluorescence (OFF), but the fluorescence turned on when incubated within reduction environment such as GSH solution. The clear fluorescence signal in tumor site was observed obviously after their intratumor injection. The nanoprobes also revealed efficient MRI contrast enhancement both in vitro and in vivo. And they showed good biocompatibility and did not demonstrate any tissue toxicity in vivo. This work gave the new possibility in designing more efficient and safer nanoprobes for future medical diagnoses.

The precise pathologic diagnosis of gastric cancer: some significant issues in clinical practice

The precise pathologic diagnosis of gastric cancer: some significant issues in clinical practice

Abstract 5650: Functional proteomics for precise cancer diagnosis and therapy using the MSK Quantitative Cancer Proteomics Atlas

Abstract The advent of molecular biology and molecular profiling in clinical medicine has transformed our understanding of the molecular basis of human cancer. As a result, we are increasingly improving the classification of human tumors based on their specific genetic and molecular mechanisms of pathogenesis. However, currently only a small number of mutant alleles guide treatment decisions, while most observed mutations remain of unknown pathologic and clinical significance. In addition, even for recently approved drugs, such as those targeting activated kinase signaling, clinical efficacy is highly varied, with no currently satisfactory means to identify molecular markers of response and resistance. To address this need, and broadly enable transformative future advances in precision oncology and patient outcomes, we have now developed the Quantitative Cancer Proteomics Atlas (QCPA). This technology permits highly multiplexed, quantitative analysis of the expression and biochemical activity of hundreds of proteins, covering most recurrently mutated and known pathogenic pathways in cancer cells, and designed to be applied to clinically accessible patient specimens. QCPA enables concomitant measurement of cellular concentration and stoichiometry of regulatory post-translational modification of key proteins mediating aberrant survival and proliferation. The activation status of these proteins is then used as a proxy to infer regulatory status of relevant biochemical pathways. These sentinel proteins include 467 known effectors of kinase regulatory signaling, apoptosis, stress response, and proliferation. QCPA measurements can be made using conventional parallel reaction monitoring mass spectrometry, or using an improved method termed accumulated ion monitoring (Cifani & Kentsis 2017). We established analytical metrics of QCPA assays in cancer cell lines, and demonstrated sensitivity and quantification accuracy amenable to measure endogenous chemically modified proteins from microgram specimens. We then demonstrated the ability of QCPA profiling to detect differential biochemical activities in response to defined stimuli in vitro. Next, we deployed QCPA profiling to reveal functional differences between primary human acute myeloid leukemia specimens and CD34+ myeloid progenitors, measuring specific activation of biochemical pathways in cancer cells. Finally, we tested the clinical utility of functional proteomic profiling by determining molecular markers of response and resistance to ERK pathway inhibitors in patient specimens. Thus, QCPA functional proteomic profiling provides a tool to achieve precise diagnosis and rational targeted cancer therapies. Citation Format: Paolo Cifani, Ahmet Dogan, David B. Solit, Alex Kentsis. Functional proteomics for precise cancer diagnosis and therapy using the MSK Quantitative Cancer Proteomics Atlas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5650.

A two-photon ratiometric probe for detection of hNQO1 enzyme activity in human colon tissue

Human colonic carcinomas are one of the most common cancers in all Western countries, but there is no reliable method for quantitative and rapid analysis of colon cancer. Herein, we report an hNQO1 enzyme-selective and two-photon emission ratiometric probe (SHC) for quantitative analysis of hNQO1 activity in human colon tissue. This probe shows a blue-to-yellow emission color change in response to hNQO1, high sensitivity and selectivity for hNQO1 over other bio-metabolites, and two-photon microscopy (TPM) imaging capability, thereby allowing precise monitoring of the activity of hNQO1 in situ. Ratiometric TPM imaging with SHC in human colon tissues revealed that hNQO1 activities gradually increased during cancer progression from normal to adenoma, and to adenocarcinoma colon tissues. These findings demonstrated that SHC labeling can detect the differential activities of hNQO1 in the distinct stages of colon neoplasm. Thus, this approach, enzyme-selective TP probe labeling, will likely provide a reliable and fast method for precise diagnosis of colon cancer.

Intrinsically Zirconium-89-Labeled Manganese Oxide Nanoparticles for In Vivo Dual-Modality Positron Emission Tomography and Magnetic Resonance Imaging.

Manganese-based nanoparticles (NPs) have recently attracted much attention in the field of biomedical imaging due to their impressive enhanced T1 contrast ability. Although the reported manganese-based NPs have exhibited good imaging capabilities as contrast agents, it is still urgent to develop novel multifunctional manganese-based imaging probes for future biomedical imaging, especially PET/MRI probes. Herein, we present chelator-free zirconium-89 (89Zr, t1/2: 78.4 h) labeling of manganese oxide NPs (Mn3O4@PEG) with ∼78% labeling yield and good stability. Serial positron emission tomography (PET) and magnetic resonance imaging (MRI) studies non-invasively assessed the biodistribution patterns of the NPs and the feasibility of in vivo dual-modality imaging and lymph-node mapping. Since Mn3O4 NPs exhibited desirable properties for enhanced T1 imaging and the simplicity of chelator-free radiolabeling, [89Zr]Mn3O4@PEG NPs offer a novel, simple, safe and accurate nanoplatforms for future precise cancer imaging and diagnosis.

On-off-on fluorescent nanosensor for Fe3+ detection and cancer/normal cell differentiation via silicon-doped carbon quantum dots

In this study, an on-off-on fluorescent nanosensor based on carbon quantum dots (or carbon dots, CDs) was successfully fabricated for Fe3+ detection and selective imaging of cancerous cells in vitro and in vivo. The multifunctional CDs were prepared by a one-pot solvothermal treatment of glycerol and a silane molecule (N-[3-(trimethoxysilyl)propyl]ethylenediamine, DAMO). The as-prepared CDs exhibited excellent fluorescence (FL) properties and favorable biocompatibility, and could realize wash-free cell imaging both in vitro (e.g., for bacterial and fungal cells) and in vivo (e.g., for zebrafish embryos). On the other hand, the fluorescence of CDs can respond to Fe3+ selectively and sensitively, with a very low detection limit of 16 nM. Besides, detection of Fe3+ in living cells and zebrafish was also successfully realized by the CDs. Moreover, it was found that glutathione (GSH) could enhance the fluorescence of the mixed solution of CDs and Fe3+ (CDs/Fe3+), and such a property could be utilized to efficiently distinguish cancerous cells from normal ones based on the difference in the content of GSH of the two types of cells. More importantly, GSH also realized the enhanced fluorescence signals of CDs/Fe3+ in tumor site in vivo after intravenous injection, indicating the potential of CDs/Fe3+ for imaging-guided precision cancer diagnosis.

Precise Diagnosis of Prostate Cancer and More

Precise Diagnosis of Prostate Cancer and More

A perylene diimide zwitterionic polymer for photoacoustic imaging guided photothermal/photodynamic synergistic therapy with single near-infrared irradiation.

Phototherapy has great promise for precise cancer diagnosis and effective therapy, but the development of one multifunctional nanoplatform for synergistic photodynamic therapy (PDT) and photothermal therapy (PTT) at a single excitation wavelength remains a challenge. In this work, a perylene diimide zwitterionic polymer PDS-PDI was synthesized via atom transfer radical polymerization (ATRP). This polymer was designed for photoacoustic imaging (PAI) guided synergistic PDT and PTT with single 660 nm near-infrared (NIR) light irradiation. The prepared PDS-PDI polymer presents high photothermal conversion efficiency (η≈ 40%) and efficient singlet oxygen quantum yield (ΦΔ≈ 16.7%) under 660 nm laser irradiation. Polymer PDS-PDI also acts as a contrast agent for PAI, offering real-time monitoring in tumor sites. Additionally, in vitro and in vivo assays indicate that polymer PDS-PDI has good biocompatibility and effective tumor destruction ability under 660 nm laser irradiation. In brief, polymer PDS-PDI prepared in this study could be applied as a dual-mode phototherapeutic agent under single laser irradiation.

Functionalized Cu3BiS3 nanoparticles for dual-modal imaging and targeted photothermal/photodynamic therapy.

Multifunctional nano-biomaterials with the integration of diagnostic and therapeutic functions have shown great promise in improving the efficacy of cancer therapy. Herein, a new nanoplatform based on functionalized Cu3BiS3 nanoparticles (NPs) is fabricated for tumour-targeted combination phototherapy. The as-synthesized hydrophobic Cu3BiS3 NPs are modified with DSPE-PEG/DSPE-PEG-NH2, followed by the conjugation of the photosensitizer chlorin e6 (Ce6) and the target ligand folic acid (FA). The introduced Ce6 can further form a chelate complex with Gd3+. The rationally designed Cu3BiS3-PEG-(Ce6-Gd3+)-FA NPs, which have high physiological stability and good biocompatibility, can specifically target FA-receptor over-expressed tumour cells. The Cu3BiS3-PEG-(Ce6-Gd3+)-FA NPs exhibit effective dual-modal CT and MR imaging in the xenografted HeLa tumours. Importantly, excellent in vivo anti-tumour effects have been achieved by synergistic photothermal/photodynamic therapy using the multifunctional NPs. We expect that this versatile nanoplatform will play a role in exploring precise cancer diagnosis and therapy.

ATP-Activatable Photosensitizer Enables Dual Fluorescence Imaging and Targeted Photodynamic Therapy of Tumor.

Targeted delivery of intracellular stimuli-activatable photosensitizers (PSs) into tumor cells to achieve selective imaging and on-demand photodynamic therapy (PDT) of tumors has provided a vital opportunity for precise cancer diagnosis and therapy. In this paper, we report a tumor targeting and adenosine triphosphate (ATP)-activatable nanophotosensitizer Apt-HyNP/BHQ2 by modifying hybrid micellar nanoparticles with both nucleolin-targeting aptamer AS1411 and quencher BHQ2-labeled ATP-binding aptamer BHQ2-ATP-apt. We demonstrated that both of the fluorescence emissions at 555 and 627 nm were quenched by BHQ2 in Apt-HyNP/BHQ2, resulting in low PDT capacity. After selective entry into tumor cells through nucleolin-mediated endocytosis, the high concentration of intracellular ATP could bind to BHQ2-ATP-apt and trigger Apt-HyNP/BHQ2 dissociation, leading to turning "on" both fluorescence and PDT. The "off-on" fluorescence emissions at both 555 and 627 nm were successfully applied for dual color fluorescence imaging of endogenous ATP levels and real-time monitoring of intracellular activation of Apt-HyNP/BHQ2 in tumor cells. Moreover, imaging-guided precise PDT of tumors in living mice was also demonstrated, allowing for selective ablation of tumors without obvious side effects. This study highlights the potential of using a combination of tumor-targeting and ATP-binding aptamers to design ATP-activatable PSs for both fluorescence imaging and imaging-guided PDT of tumors in vivo.

Targeted bisulfite sequencing identified a panel of DNA methylation-based biomarkers for esophageal squamous cell carcinoma (ESCC)

BackgroundDNA methylation has been implicated as a promising biomarker for precise cancer diagnosis. However, limited DNA methylation-based biomarkers have been described in esophageal squamous cell carcinoma (ESCC).MethodsA high-throughput DNA methylation dataset (100 samples) of ESCC from The Cancer Genome Atlas (TCGA) project was analyzed and validated along with another independent dataset (12 samples) from the Gene Expression Omnibus (GEO) database. The methylation status of peripheral blood mononuclear cells and peripheral blood leukocytes from healthy controls was also utilized for biomarker selection. The candidate CpG sites as well as their adjacent regions were further validated in 94 pairs of ESCC tumor and adjacent normal tissues from the Chinese Han population using the targeted bisulfite sequencing method. Logistic regression and several machine learning methods were applied for evaluation of the diagnostic ability of our panel.ResultsIn the discovery stage, five hyper-methylated CpG sites were selected as candidate biomarkers for further analysis as shown below: cg15830431, P = 2.20 × 10−4; cg19396867, P = 3.60 × 10−4; cg20655070, P = 3.60 × 10−4; cg26671652, P = 5.77 × 10−4; and cg27062795, P = 3.60 × 10−4. In the validation stage, the methylation status of both the five CpG sites and their adjacent genomic regions were tested. The diagnostic model based on the combination of these five genomic regions yielded a robust performance (sensitivity = 0.75, specificity = 0.88, AUC = 0.85). Eight statistical models along with five-fold cross-validation were further applied, in which the SVM model reached the best accuracy in both training and test dataset (accuracy = 0.82 and 0.80, respectively). In addition, subgroup analyses revealed a significant difference in diagnostic performance between the alcohol use and non-alcohol use subgroups.ConclusionsMethylation profiles of the five genomic regions covering cg15830431 (STK3), cg19396867, cg20655070, cg26671652 (ZNF418), and cg27062795 (ZNF542) can be used for effective methylation-based testing for ESCC diagnosis.

Radiolabeled, Antibody-Conjugated Manganese Oxide Nanoparticles for Tumor Vasculature Targeted Positron Emission Tomography and Magnetic Resonance Imaging.

Manganese oxide nanoparticles (Mn3O4 NPs) have attracted a great deal of attention in the field of biomedical imaging because of their ability to create an enhanced imaging signal in MRI as novel potent T1 contrast agents. In this study, we present tumor vasculature-targeted imaging in mice using Mn3O4 NPs through conjugation to the anti-CD105 antibody TRC105 and radionuclide copper-64 (64Cu, t1/2: 12.7 h). The Mn3O4 conjugated NPs, 64Cu-NOTA-Mn3O4@PEG-TRC105, exhibited sufficient stability in vitro and in vivo. Serial positron emission tomography (PET) and magnetic resonance imaging (MRI) studies evaluated the pharmacokinetics and demonstrated targeting of 64Cu-NOTA-Mn3O4@PEG-TRC105 to 4T1 murine breast tumors in vivo, compared to 64Cu-NOTA-Mn3O4@PEG. The specificity of 64Cu-NOTA-Mn3O4@PEG-TRC105 for the vascular marker CD105 was confirmed through in vivo, in vitro, and ex vivo experiments. Since Mn3O4 conjugated NPs exhibited desirable properties for T1 enhanced imaging and low toxicity, the tumor-specific Mn3O4 conjugated NPs reported in this study may serve as promising multifunctional nanoplatforms for precise cancer imaging and diagnosis.

Bioinformatics-Based Identification of Methylated-Differentially Expressed Genes and Related Pathways in Gastric Cancer.

The aim of the study was to identify methylated-differentially expressed genes (MDEGs) in gastric cancer and investigate their potential pathways. Expression profiling (GSE13911 and GSE29272) and methylation profiling (GSE25869 and GSE30601) data were obtained from GEO DataSets. Differentially expressed genes and differentially methylated genes were identified using GEO2R. Gene ontology and pathway enrichment analyses were performed for the MDEGs. Protein-protein interaction (PPI) networks were established by STRING and Cytoscape. Analysis of modules in the PPI networks was performed using MCODE. Further, the hub genes derived from the PPI networks were verified by The Cancer Genome Atlas (TCGA) database and human tissues, with methylation-specific PCR for genes methylation and real-time qPCR for genes expression. A total of 445 genes were identified as hypermethylated, lowly expressed genes (Hyper-LGs), which were enriched in the regulation of system process and channel activity. A total of 129 genes were identified as hypomethylated, highly expressed genes (Hypo-HGs), which were involved in cell adhesion, cell proliferation, and protein binding. Pathway analysis showed that Hyper-LGs were associated with neuroactive ligand-receptor interaction and calcium signaling pathway, while Hypo-HGs were enriched in pathways in cancer. In the PPI networks, after verification by TCGA analysis and human tissue detection, CASR, CXCL12, and SST were identified as significantly different hub genes. MDEG analysis helps to understand the epigenetic regulation mechanisms involved in the development and progression of gastric cancer. The hub genes have predictive and prognostic value as methylation-based biomarkers for the precise diagnosis and treatment of gastric cancer.

Down-regulation of traditional oncomiRs in plasma of breast cancer patients.

Deregulated expression of microRNAs has the oncogenic or tumor suppressor function in cancer. Since miRNAs in plasma are highly stable, their quantification could contribute to more precise cancer diagnosis, prognosis and therapy prediction. We have quantified expression of seven oncomiRs, namely miR-17/92 cluster (miR-17, miR-18a, miR-19a and miR-20a), miR-21, miR-27a and miR-155, in plasma of 137 breast cancer (BC) patients. We detected down-regulation of six miRNAs in patients with invasive BC compared to controls; however, only miR-20a and miR-27a down-regulations were statistically significant. Comparing miRNA expression between early and advanced stages of BC, we observed statistically significant decrease of miR-17 and miR-19a. We identified down-regulation of miR-17 and miR-20a in patients with clinical parameters of advanced BC (lymph node metastasis, tumor grade 3, circulating tumor cells, higher Ki-67-related proliferation, hormone receptor negativity and HER2 amplification), when compared to controls. Moreover, decreased level of miR-17 was found from low to high grade. Therefore, miR-17 could represent an indicator of advanced BC. Down-regulated miR-27a expression levels were observed in all clinical categories regardless of tumor progression. Hence, miR-27a could be used as a potential diagnostic marker for BC. Our data indicates that any changes in miRNA expression levels in BC patients in comparison to controls could be highly useful for cancer-associated pathology discrimination. Moreover, dynamics of miRNA expression changes could be used for BC progression monitoring.

Multi-Förster Resonance Energy Transfer-Based Fluorescent Probe for Spatiotemporal Matrix Metalloproteinase-2 and Caspase-3 Imaging.

A novel single-molecular fluorescent probe was developed for spatiotemporal matrix metalloproteinase-2 (MMP-2) and caspase-3 imaging with distinct fluorescence signals. Due to the multi-Förster resonance energy transfer (FRET) processes, the probe could respond to MMP-2 and caspase-3 independently with high signal-to-noise ratio. Moreover, the overexpression of MMP-2 in cancer cell lines and the cisplatin induced cell apoptosis were spatiotemporal imaged with distinct fluorescence emissions. Because of the independent process of the probe for MMP-2 and caspase-3 imaging, the probe could meet the demands for precise disease diagnosis and cancer theranostic applications, which could extensively simplify the processes for precise cancer diagnosis and imaging.

Targeted nanomedicine for cancer therapeutics: Towards precision medicine overcoming drug resistance.

Intrinsic anticancer drug resistance appearing prior to chemotherapy as well as acquired resistance due to drug treatment, remain the dominant impediments towards curative cancer therapy. Hence, novel targeted strategies to overcome cancer drug resistance constitute a key aim of cancer research. In this respect, targeted nanomedicine offers innovative therapeutic strategies to overcome the various limitations of conventional chemotherapy, enabling enhanced selectivity, early and more precise cancer diagnosis, individualized treatment as well as overcoming of drug resistance, including multidrug resistance (MDR). Delivery systems based on nanoparticles (NPs) include diverse platforms enabling a plethora of rationally designed therapeutic nanomedicines. Here we review NPs designed to enhance antitumor drug uptake and selective intracellular accumulation using strategies including passive and active targeting, stimuli-responsive drug activation or target-activated release, triggered solely in the cancer cell or in specific organelles, cutting edge theranostic multifunctional NPs delivering drug combinations for synergistic therapy, while facilitating diagnostics, and personalization of therapeutic regimens. In the current paper we review the recent findings of the past four years and discuss the advantages and limitations of the various novel NPs-based drug delivery systems. Special emphasis is put on in vivo study-based evidences supporting significant therapeutic impact in chemoresistant cancers. A future perspective is proposed for further research and development of complex targeted, multi-stage responsive nanomedical drug delivery systems for personalized cancer diagnosis and efficacious therapy.

Artificial Intelligence Techniques for Cancer Detection and Classification: Review Study

Cancer is the general name for a group of more than 100 diseases. Although cancer includes different types of diseases, they all start because abnormal cells grow out of control. Without treatment, cancer can cause serious health problems and even loss of life. Early detection of cancer may reduce mortality and morbidity. This paper presents a review of the detection methods for lung, breast, and brain cancers. These methods used for diagnosis include artificial intelligence techniques, such as support vector machine neural network, artificial neural network, fuzzy logic, and adaptive neuro-fuzzy inference system, with medical imaging like X-ray, ultrasound, magnetic resonance imaging, and computed tomography scan images. Imaging techniques are the most important approach for precise diagnosis of human cancer. We investigated all these techniques to identify a method that can provide superior accuracy and determine the best medical images for use in each type of cancer.

Novel iodinated gold nanoclusters for precise diagnosis of thyroid cancer.

As the most common endocrine malignancy with a high incidence, thyroid cancer lacks a dual-modal imaging method for precise diagnosis. An accurate and multimodal imaging system is pivotal to solve this problem. Herein, dual-modality fluorescence/Computed Tomography (CT) iodinated gold nanoclusters for malignant thyroid cancer visualization have been recently fabricated. In this study, innovative iodinated gold nanoclusters (AuNCs@BSA-I) were synthesized via Bovine serum albumin (BSA) and chloramine-T. AuNCs@BSA-I not only possess an ultra-small size and brilliant biocompatibility but also exhibit excellent fluorescence/CT imaging properties. Particularly with regard to CT imaging properties, AuNCs@BSA-I rival the clinical CT contrast medium. And the fluorescence emission spectrum of AuNCs@BSA-I falls in the near infrared region (NIR). For further translational application in medicine, we established an orthotopic human thyroid cancer patient tissue derived xenograft (PDX) mouse model, highly close to the actual human thyroid cancer. The results unveil that AuNCs@BSA-I exert sensitive and accurate diagnosis characteristics. To be more specific, the AuNCs@BSA-I fluorescent/CT signals in the thyroid tumor represent characteristics of 'slow in fast out', compared to those in the normal thyroid. Moreover, AuNCs@BSA-I could distinguish minimal thyroid cancer, as small as 2 mm3. Therefore, AuNCs@BSA-I appear to be a promising nanoprobe which could be applied to preclinical medicine.

Isolation and retrieval of circulating tumor cells on a microchip with double parallel layers of herringbone structure

Isolation of the extremely rare circulating tumor cells (CTCs) from peripheral blood has become an effective tool for monitoring and staging tumor development, based on which the therapeutic efficacy can be evaluated. Many techniques have been developed for CTC isolation and enumeration. However, isolation and detection of CTCs with high sensitivity remains a challenging work, and there still exists an urgent demand to retrieve tumor cells after isolation for further molecular and cellular analyses. Herein, we report a dual-functional microchip with double parallel layers of herringbone structure. CTC isolation and retrieving can be simply achieved based on the reversible conjugation with anti-EpCAM modified on the interior channel surface. Specifically, two major strategies were engaged to improve the CTC isolation based on the synergistic effect of significantly increased functional surface area and local microvortex mixing. This approach could achieve CTC isolation efficiency of 75% on average and the detection limit down to 10 cells mL−1. Furthermore, CTC retrieving rate was found at 73.4%. Therefore, the present method has a good potential for quantitation and retrieving CTCs with high efficiency for precise cancer diagnosis and subsequent cellular and molecular analyses.