Enrichment and detection of circulating tumor cells in peripheral blood

Metastasis usually presents in the different courses of tumor diseases and associates with poor prognosis.Circulating tumor cells(CTC) are considered to be essential for conforming metastasis and can be detected in tumor patients' peripheral blood.CTC are not easily detected by conventional cytology methods because of their low frequency in peripheral blood.Our article reviews the recent research results on this subject,and also discusses the problems and prospects in this area.

Clinical Value of Peripheral Blood Circulating Tumor Cells and Cell-Free DNA Combined Detection in Triple-Negative Breast Cancer

Objective:To determine the clinical value of combined detection of circulating tumor cells(CTCs)and cell-free DNA(cfDNA)in peripheral blood of patients with triple-negative breast cancer.Method:41 patients with breast cancer admitted to the First Central Hospital of Baoding from January 2020 to December 2021 were selected and recruited into the experimental group,42 patients with benign breast cancer admitted during the same period were recruited into the conditional control group,and 41 healthy patients admitted during the same period were recruited into the blank control group.The positive rate of peripheral blood CTCs,the level of cfDNA,and the diagnostic efficacy of peripheral blood CTCs,cfDNA alone and the combination thereof for breast cancer were analyzed.Result:The positive rates of peripheral blood CTCs in the experimental group,the conditional control group,and the blank control group were 43.90%,11.90%,and 9.74%,respectively,and there was significant difference among the groups.The levels of cfDNA in peripheral blood of the experimental group,the conditional control group,and the blank control group were 0.26±0.08 bp,0.17±0.03 bp,and 0.15±0.04 bp,respectively,which were statistically significant.The detection levels of 100 bp hTERT/ng mT1 and 241 bp hTERT/ng-mT1 in the experimental group were significantly higher than those in the conditional control group and the blank control group.The accuracy of peripheral blood CTCs detection in the three groups was 66.21%,the accuracy of cfDA241 bp/100 hp hTERT detection was 80.41%,and the accuracy of combined detection of peripheral blood CTCs and cfDNA was 94.03%.Conclusion:The clinical application of peripheral blood CTCs combined with cfDNA level detection can increase detection accuracy,provide data support for clinicians,and improve the clinical diagnostic effect of triple-negative breast cancer.

Application of Circulating Tumor Cells in Peripheral Blood in Judging the Prognosis of Patients with Renal Cancer and Related Indexes of Blood Coagulation

Objective: To investigate the value of the number of circulating tumor cells (CTC) in peripheral blood in the prognosis and coagulation-related indicators of patients with renal cancer. Methods: 65 patients with renal cell carcinoma (RCC) confirmed pathologically were divided into CTC positive group and CTC negative group according to the CTC count (5 pcs/3.5 ml). Compare the age, gender, tumor location, TNM (clinical stage), pathological grade, tissue type, lymph node metastasis, distant metastasis, prognosis and prothrombin time (PT), fibrinogen (FIB), partial coagulation of the two groups of patients The correlation between the results of zymogen time (APTT) and D-dimer (DD) and the number of CTC. Results: There were significant differences in TNM, lymph node metastasis, and distant metastasis between the two groups (P < 0.05). The number of CTC in patients was correlated with FIB and D-D levels (P < 0.05). Conclusion: The number of CTC in patients with renal cell carcinoma is correlated with some clinical phenotypes (TNM, lymph node metastasis, distant metastasis) and some coagulation indexes (FIB, D-D), and can jointly predict the prognosis of renal cancer.

Advances in isolation and detection of circulating tumor cells based on microfluidics

Circulating tumor cells(CTCs) are the cancer cells that circulate in the peripheral blood after escaping from the original or metastatic tumors. CTCs could be used as non-invasive source of clinical information in early diagnosis of cancer and evaluation of cancer development. In recent years, CTC research has become a hotspot field wherein many novel CTC detection technologies based on microfluidics have been developed. Great advances have been made that exhibit obvious technical advantages, but cannot yet satisfy the current clinical requirements. In this study, we review the main advances in isolation and detection methods of CTC based on microfluidics research over several years, propose five technical indicators for evaluating these methods, and explore the application prospects. We also discuss the concepts, issues, approaches, advantages, limitations, and challenges with an aim of stimulating a broader interest in developing microfluidics-based CTC detection technology.

Detection of Circulating Tumor Cells in Patients with Breast, Prostate, Pancreatic, Colon and Melanoma Cancer: A Blinded Comparative Study Using Healthy Donors

Cancer is a diverse disease characterized by abnormal cell growth and the ability to invade or spread to other parts of the body. Because the yearly cancer rate is increasing, an important area for cancer researchers is to improve the ability to detect and treat cancer early. The current study analyzes the potential of flow cytometry to be used to detect circulating tumor cells (CTCs) in patients with various cancer types and stages. CTCs are cells that have detached from the primary tumor and entered the blood stream in the process of metastasizing to other organs. To determine the accuracy of flow cytometry in detecting CTCs, a comparative study was performed on healthy donors. In this study, blood samples from patients with breast, prostate, pancreatic, colon and skin cancer were analyzed and compared with healthy donors. The data were collected and analyzed statistically with receiver operating characteristic curve analysis. The results indicate that CTCs can be detected in over 83% of the cancer patients and therefore may be a promising method for diagnosing cancer.

A Correlation Analysis of Postoperative Hypercoagulability and Peripheral Circulating Tumor Cells in Patients with Lung Cancer

Objective:To explore the correlation between peripheral circulating tumor cells and hypercoagulability in patients with lung cancer after surgery.Methods:From January 2017 to December 2021,89 patients with lung cancer who were treated in the Affiliated Hospital of Hebei University were selected as the research subjects,and a retrospective analysis was conducted to analyze and observe the D-dimer(DD),fibrinogen(FIB),and platelet(PLT)levels in peripheral blood,as well as detect peripheral CTC.Results:There were statistical differences in TMN staging,tumor metastasis,and lymph node metastasis in the clinical data,but there were no statistical differences in gender,smoking history,and pathological classification.After retrospective analysis and comparison of the patients,the DD(mg/ml),FIB(g/L),and PLT(×10^(9)/L)levels of the CTC positive group were 3.41±0.58,3.98±0.87,and 367.26±34.98,respectively;the CTC negative group’s DD(mg/ml),FIB(g/L),and PLT(×10^(9)/L)levels were 0.89±0.49,1.06±0.45,and 234.69±35.69,respectively,and the differences were statistically significant.The factors affecting the prognosis of patients included TMN staging and CTC;the number of CTC positives in the death group was significantly higher than that in the survival group,and there was a statistical difference between the groups.Gender,age,smoking history,pathological type,and surgical resection had no effect on the prognosis of patients.Among the enrolled patients,the survival rate was 71.91%.Conclusion:CTC-positive patients have a higher probability of hypercoagulability after surgery and are prone to tumor metastasis;thus,CTC can be used as a judgment index for the prognosis of patients.

Relationship between Single Nucleotide Polymorphism in TNF-α Gene Promoter Region and Inhibitory Effects of Triptolide on TNF-α Production in Peripheral Blood Mononuclear Cells of Healthy Humans

The relationship between tumour necrosis lactose （TNF-α） gene polymorphism and inhibitory effects of triptolide on TNF-α production from peripheral blood mononuclear cells （PBMC） of healthy humans was investigated. Genomic DNA from 41 healthy people was typed for TNF-α- 308 polymorphism by allele-specific polymorphism chain reaction （AS-PCR）. The TNF-α concentration in the supernatant was measured by ELISA. The results showed that the production of TNF-α from TNF-α -308 non-G/G genotype PBMC was higher than that from TNF-α-308 G/G genotype PBMC after stimulated by LPS. Triptolide could lower the production of TNF-α from G/ G genotype PBMC, but had no effect on the level of TNF-α from non-G/G genotype PBMC. It was concluded that TNF-α gene polymorphism was related to the TNF-α production from triptolide-inhibited PBMC culture in healthy humans.

Early detection of circulating tumor DNA and successful treatment with osimertinib in thr790met-positive leptomeningeal metastatic lung cancer:A case report

BACKGROUND Patients diagnosed with non-small-cell lung cancer with activated epidermal growth factor receptor mutations are more likely to develop leptomeningeal(LM)metastasis than other types of lung cancers and have a poor prognosis.Early diagnosis and effective treatment of leptomeningeal carcinoma can improve the prognosis.CASE SUMMARY A 55-year-old female with a progressive headache and vomiting for one month was admitted to Peking University First Hospital.She was diagnosed with lung adenocarcinoma with osseous metastasis 10 months prior to admittance.epidermal growth factor receptor(EGFR)mutation was detected by genomic examination,so she was first treated with gefitinib for 10 months before acquiring resistance.Cell-free cerebrospinal fluid(CSF)circulating tumor DNA detection by next-generation sequencing was conducted and indicated the EGFR-Thr790Met mutation,while biopsy and cytology from the patient’s CSF and the first enhanced cranial magnetic resonance imaging(MRI)showed no positive findings.A month later,the enhanced MRI showed linear leptomeningeal enhancement,and the cytology and biochemical examination in CSF remained negative.Therefore,osimertinib(80 mg/d)was initiated as a second-line treatment,resulting in a good response within a month.CONCLUSION This report suggests clinical benefit of osimertinib in LM patients with positive detection of the EGFR-Thr790Met mutation in CSF and proposes that the positive findings of CSF circulating tumor DNA as a liquid biopsy technology based on the detection of cancer-associated gene mutations may appear earlier than the imaging and CSF findings and may thus be helpful for therapy.Moreover,the routine screening of chest CT with the novel coronavirus may provide unexpected benefits。

Circulating tumor cells in lung cancer:Detection methods and clinical impact

Circulating tumor cells(CTCs) are tumor cells that enter the blood circulation after detaching from the primary tumor and can migrate to reach distant organs, where they can give rise to aggressive metastasis. Clinical studies have revealed that the presence of CTCs in peripheral blood is correlated with disease progression in lung cancer. However, as CTCs are rare cancer cells released from tumors into the bloodstream, both enrichment and sensitive detection methods are technically challenging. In order to best understand how CTCs are currently being deployed, this review mainly focuses on the different detection methods for CTCs. Furthermore, we will describe the clinical impact of circulating tumor cells in lung cancer and discuss their potential use as biomarker to guide the prognosis.

Effect of Shenqi Fuzheng Injection combined with chemotherapy on peripheral blood cell level, tumor markers and immune function in patients with gastric cancer

Objective:To investigate the effects of Shenqi Fuzheng Injection combined with chemotherapy on peripheral blood cell level, tumor markers and immune function in patients with gastric cancer.Methods: Eighty patients with gastric cancer admitted to our hospital from May 2016 to October 2017 were selected as the research object and randomly divided into observation group and control group, 40 cases in each group. Control group was treated by DCF chemotherapy, while the observation group by Shenqi Fuzheng Injection based on the DCF chemotherapy. The levels of peripheral blood cells, tumor markers and immune function indexes in both groups were detected and compared before and after treatment.Results: There were no significant differences in serum RBC, WBC, PLT, Hb, NSE, CYFRA21-1, CA199, CEA, CD3+, CD4+, CD8+ and CD4+/CD8+ levels before treatment in both groups. Compared with the pretreatment group, the levels of RBC, WBC, PLT and Hb in both groups decreased to some extent after treatment, and the levels in the observation group were significantly higher than those in the control group;Compared with the same group before treatment, the levels of NSE, CYFRA21-1, CA199 and CEA in the two groups were significantly decreased, and the observation group was significantly lower than the control group;CD3+, CD4+, CD4+/CD8+ levels in control group and observation group increased to some extent after treatment, and the levels in observation group were significantly higher than those in control group. The levels of CD8+ in both groups after treatment were significantly lower than those before treatment, and the observation group was significantly lower than the control group after treatment. The above data for statistical analysis had significant differences.Conclusions: Shenqi Fuzheng injection combined with chemotherapy helps to clear and kill cancer cells, enhance immunity and reduce hematological toxicity after chemotherapy, which can be used as an adjuvant chemotherapy for gastric cancer.

Influence of granulocyte-macrophage colonystimulating factor and tumor necrosis factor on anti-hepatoma activities of human dendritic cells

INTRODUCTIONDendritic cells (DCs) play a key regulatory role inantitumor immunity,especially in its immuneaccessory role via MHC-Ⅰ molecules.We haverecently reported that DCs were able to enhance thekilling activity of Lymphokine and PHA activatedkiller (LPAK) cells in vitro.In the presentstudy,we evaluated the effects of GM-CSF andTNF upon antitumor activities of freshly

Squamous cell carcinoma of the oral cavity and circulating tumour cells

Due to a lack of substantial improvement in the outcome of patients suffering from oral squamous cell carcinoma(OSCC) during the past decades, current staging methods need to be revised. This disease is associated with poor survival rates despite considerable advances in diagnosis and treatment. The early detection of metastases is an important indicator of survival, prognosis and relapse. Therefore, a better understanding of the mechanisms underlying metastasis is crucial. Exploring alternative measures apart from common procedures is needed to identify new prognostic markers. Similar to previous findings predominantly for other solid tumours, recently published studies demonstrate that circulating tumour cells(CTCs) and disseminated tumour cells(DTCs) might serve as prognostic markers and could supplement routine staging in OSCC. Thus, the detection of CTCs/DTCs is a promising tool todetermine the individual need for therapeutic intervention. Encouraging results and new approaches point to the future use of targeted therapies for OSCC, an exceedingly heterogeneous subgroup of head and neck cancer. This review focuses on summarising technologies currently used to detect CTCs/DTCs. The translational relevance for OSCC is highlighted. The inherent challenges in detecting CTCs/DTCs will be emphasised.

Lactobacilli,bifi dobacteria and E.coli nissle induce pro-and anti-inflammatory cytokines in peripheral blood mononuclear cells

AIM： To investigate whether the stimulation of peripheral blood mononuclear cells （PBMNC） with the cell debris and cell extraction of different probiotic strains is similar or species specific. METHODS： Three strains of bifidobacteria, 4 strains of lactobacilli, and E. coli nissle were sonicated and centrifuged in order to divide them into cell extract and cell debris. PBMNC were separated by density gradient and incubated for 36 h with either the cell debris or the cell extract of single strains of probiotic bacteria in doses from 10^2 to 10^8 CFU/mL. Cell supernatants were taken and interleukin （IL）-10, IL-1β, and tumor necosis factor （TNF）-α were determined by ELISA. RESULTS： Depending on the species super-family, the strains had different stimulation patterns. Except for both L. casei strains, the cell extract of bitTdobacteria and/actobacilli had less stimulating capacity than cell debris, whereas the cell extract of E. coli nissle had similar stimulating properties to that of the cell debris of the strain and significantly more stimulating capacity than that of bifidobacteria and lactobacilli. The cell debris of bifidobacteria stimulated more cytokine release than the cell debris of lactobacilli. The cell debris of lactobacilli did not have a stimulating capacity when lower concentrations were used. Neither cell extraction nor cell debris had an inhibitory effect on the production of the tested cytokines by stimulated PBMNC. CONCLUSION： The incubation of probiotic strains, which have been used in clinical trials for inflammatory diseases, with immunocompetent cells leads to different species specific reactions. High IL-10 response to cell debris of bifidobacteria and E. coli nissle can be found. This corresponds to positive effects of bihdobacteria and E. coli nissle in clinical trials for inflammatory bowel disease compared to negative outcomes obtained with lactobacilli.

Cultured circulating tumor cells and their derived xenografts for personalized oncology

Recent cancer research has demonstrated the existence of circulating tumor cells(CTCs)in cancer patient’s blood.Once identified,CTC biomarkers will be invaluable tools for clinical diagnosis,prognosis and treatment.In this review,we propose ex vivo culture as a rational strategy for large scale amplification of the limited numbers of CTCs from a patient sample,to derive enough CTCs for accurate and reproducible characterization of the biophysical,biochemical,gene expressional and behavioral properties of the harvested cells.Because of tumor cell heterogeneity,it is important to amplify all the CTCs in a blood sample for a comprehensive understanding of their role in cancer metastasis.By analyzing critical steps and technical issues in ex vivo CTC culture,we developed a cost-effective and reproducible protocol directly culturing whole peripheral blood mononuclear cells,relying on an assumed survival advantage in CTCs and CTC-like cells over the normal cells to amplify this specified cluster of cancer cells.

Circulating cytokeratin-positive cells and tumor budding in colorectal cancer

AIM To investigate whether circulating cytokeratin-positive(CK^+) cells in the mesenteric blood of resected colorectal specimens are prognostic and correlate with tumor budding.METHODS Fifty-six colorectal specimens were collected between 9/2007 and 7/2008.Blood from the mesenteric vein was drawn immediately after receiving the fresh and unfixed specimens in the pathology department.After separation of the mononuclear cells by Ficoll-Hypaquedensity-gradient centrifugation,cytological smears were immunocytochemically stained for CK18.Tumor budding was evaluated on slides stained for pan-cytokeratin.The identification of ≥ 30 buds/1.3 mm2 was defined as high grade budding.RESULTS CK^+ cells and clusters were identified in 29(48%) and 14(25%) of the samples,respectively.Two cells were identified in one of three non-malignant cases.Clusters were found exclusively in malignant cases.The occurrence of CK^+ cells or clusters was not associated with any of the evaluated clinicopathological factors,including surgical technique and tumor budding.Moreover,the occurrence of CK^+ cells or clusters had no influence on the cancerspecific survival [75 mo(CI:61;88) vs 83 mo(CI:72;95) and 80 mo(CI:63;98) vs 79 mo(CI:69;89),respectively].CONCLUSION CK^+ cells and showed neither prognostic significance nor an association with tumor budding.It is very likely that CK18-staining is not specific enough to identify the relevant cells.

On-chip circulating tumor cells isolation based on membrane filtration and immuno-magnetic bead clump capture

Isolating rare circulating tumor cells(CTCs)from blood is critical for the downstream analysis that is important in cancer-related research,diagnosis,and medicine,and efforts are ongoing to increase the efficiency and purity of CTC isolation in microfluidics.Reported in this paper is a two-stage integrated microfluidic chip for coarse-to-fine CTC isolation from whole blood.First,blood cells are removed by filtration using a micropore-array membrane,then CTCs and other cells that are trapped in the micropores are peeled off the membrane by a novel release method based on air–liquid interfacial tension,which significantly increases the recovery rate of CTCs.The second stage is CTC capture based on an on-chip dense immuno-magnetic-bead clump,which offers high capture efficiency and purity.Both the micropore filtration and immuno-magnetic-bead capture are validated and optimized experimentally.Overall,the integrated microfluidic chip can realize a recovery rate of 85.5%and a purity of 37.8%for rare cancer cells spiked in whole blood.

Establishment of an optimized CTC detection model consisting of EpCAM,MUC1 and WT1 in epithelial ovarian cancer and its correlation with clinical characteristics

Objective:Emerging studies have demonstrated the promising clinical value of circulating tumor cells(CTCs)for diagnosis,disease assessment,treatment monitoring and prognosis in epithelial ovarian cancer.However,the clinical application of CTC remains restricted due to diverse detection techniques with variable sensitivity and specificity and a lack of common standards.Methods:We enrolled 160 patients with epithelial ovarian cancer as the experimental group,and 90 patients including 50 patients with benign ovarian tumor and 40 healthy females as the control group.We enriched CTCs with immunomagnetic beads targeting two epithelial cell surface antigens(EpCAM and MUC1),and used multiple reverse transcription-polymerase chain reaction(RT-PCR)detecting three markers(EpCAM,MUC1 and WT1)for quantification.And then we used a binary logistic regression analysis and focused on EpCAM,MUC1 and WT1 to establish an optimized CTC detection model.Results:The sensitivity and specificity of the optimized model is 79.4%and 92.2%,respectively.The specificity of the CTC detection model is significantly higher than CA125(92.2%vs.82.2%,P=0.044),and the detection rate of CTCs was higher than the positive rate of CA125(74.5%vs.58.2%,P=0.069)in early-stage patients(stage I and II).The detection rate of CTCs was significantly higher in patients with ascitic volume≥500 mL,suboptimal cytoreductive surgery and elevated serum CA125 level after 2 courses of chemotherapy(P<0.05).The detection rate of CTC;and CTC;was significantly higher in chemo-resistant patients(26.3%vs.11.9%;26.4%vs.13.4%,P<0.05).The median progression-free survival time for CTC;patients trended to be longer than CTC;patients,and overall survival was shorter in CTC;patients(P=0.043).Conclusions:Our study presents an optimized detection model for CTCs,which consists of the expression levels of three markers(EpCAM,MUC1 and WT1).In comparison with CA125,our model has high specificity and demonstrates better diagnostic values,especially for early-stage ovarian cancer.Detection of CTC;and CTC;had predictive value for chemotherapy resistance,and the detection of CTC;suggested poor prognosis.

Detecting circulating tumor material and digital pathology imaging during pancreatic cancer progression

Pancreatic cancer(PC) is a leading cause of cancerrelated death worldwide. Clinical symptoms typically present late when treatment options are limited and survival expectancy is very short. Metastatic mutations are heterogeneous and can accumulate up to twenty years before PC diagnosis. Given such genetic diversity, detecting and managing the complex states of disease progression may be limited to imaging modalities and markers present in circulation. Recent developments in digital pathology imaging show potential for early PC detection, making a differential diagnosis, and predicting treatment sensitivity leading to long-term survival in advanced stage patients. Despite large research efforts, the only serum marker currently approved for clinical use is CA 19-9. Utility of CA 19-9 has been shown to improve when it is used in combination with PC-specific markers. Efforts are being made to develop early-screening assays that can detect tumor-derived material, present in circulation, before metastasis takes a significant course. Detection of markers that identify circulating tumor cells and tumor-derived extracellular vesicles(EVs) in biofluid samples offers a promising non-invasive method for this purpose. Circulating tumor cells exhibit varying expression of epithelial and mesenchymal markers depending on the state of tumor differentiation. This offers a possibility for monitoring disease progression using minimally invasive procedures. EVs also offer the benefit of detecting molecular cargo of tumor origin and add the potential to detect circulating vesicle markers from tumors that lack invasive properties. This review integrates recent genetic insights of PC progression with developments in digitalpathology and early detection of tumor-derived circulating material.

Establishment of a humanized mouse model using steady-state peripheral blood-derived hematopoietic stem and progenitor cells facilitates screening of cancer-targeted T-cell repertoires

Background:Cancer-targeted T-cell receptor T(TCR-T)cells hold promise in treating cancers such as hematological malignancies and breast cancers.However,approaches to obtain cancer-reactive TCR-T cells have been unsuccessful.Methods:Here,we developed a novel strategy to screen for cancer-targeted TCR-T cells using a special humanized mouse model with person-specific immune fingerprints.Rare steady-state circulating hematopoietic stem and progenitor cells were expanded via three-dimensional culture of steady-state peripheral blood mononuclear cells,and then the expanded cells were applied to establish humanized mice.The human immune system was evaluated according to the kinetics of dendritic cells,monocytes,T-cell subsets,and cytokines.To fully stimulate the immune response and to obtain B-cell precursor NAML-6-and triple-negative breast cancer MDA-MB-231-targeted TCR-T cells,we used the inactivated cells above to treat humanized mice twice a day every 7 days.Then,human T cells were processed for TCRβ-chain(TRB)sequencing analysis.After the repertoires had been constructed,features such as the fraction,diversity,and immune signature were investigated.Results:The results demonstrated an increase in diversity and clonality of T cells after treatment.The preferential usage and features of TRBV,TRBJ,and the V–J combination were also changed.The stress also induced highly clonal Science and Technology,Grant/Award Number:2021C03010;Zhejiang Provincial Natural Science Foundation of China,Grant/Award Numbers:LTGY24H080003,LY21H080004 expansion.Tumor burden and survival analysis demonstrated that stress induction could significantly inhibit the growth of subsequently transfused live tumor cells and prolong the survival of the humanized mice.Conclusions:We constructed a personalized humanized mouse model to screen cancer-targeted TCR-T pools.Our platform provides an effective source of cancer-targeted TCR-T cells and allows for the design of patient-specific engineered T cells.It therefore has the potential to greatly benefit cancer treatment.

Antibody-engineered red blood cell interface for high-performance capture and release of circulating tumor cells

Circulating tumor cells(CTCs),as important liquid biopsy target,can provide valuable information for cancer progress monitoring and individualized treatment.However,current isolation platforms incapable of balancing capture efficiency,specificity,cell viability,and gentle release have restricted the clinical applications of CTCs.Herein,inspired by the structure and functional merits of natural membrane interfaces,we established an antibody-engineered red blood cell(RBC-Ab)affinity interface on microfluidic chip for high-performance isolation and release of CTCs.The lateral fluidity,pliability,and anti-adhesion property of the RBC microfluidic interface enabled efficient CTCs capture(96.5%),high CTCs viability(96.1%),and high CTCs purity(average 4.2-log depletion of leukocytes).More importantly,selective lysis of RBCs by simply changing the salt concentration was utilized to destroy the affinity interface for efficient and gentle release of CTCs without nucleic acid contamination.Using this chip,CTCs were successfully detected in colon cancer samples with 90%sensitivity and 100%specificity(20 patients and 10 healthy individuals).After the release process,KRAS gene mutations of CTCs were identified from all the 5 cancer samples,which was consistent with the results of tissue biopsy.We expect this RBC interface strategy will inspire further biomimetic interface construction for rare cell analysis.