Positive Rate Of Circulating Tumor Cells Research Articles

The Presence of Small-Size Circulating Tumor Cells Predicts Worse Prognosis in Non-Small Cell Lung Cancer Patients.

Most patients with non-small cell lung cancers (NSCLC) are diagnosed at advanced stages. The 5-year survival rate of patients with advanced lung cancer is less than 20%, which makes lung cancer the leading cause of cancer-related deaths worldwide. To identify indicators that can predict the prognosis of lung cancer patients. To determine the correlation between circulating tumor cells (CTCs), circulating tumor-derived endothelial cells (CTECs), and their subtypes and the prognosis of patients with NSCLC, 80 patients with lung cancer were recruited and 48 patients who met the enrollment criteria were selected in this study. Peripheral blood was collected from the enrolled patients before any treatment and analyzed by the subtraction enrichment and immunostaining-fluorescence in situ hybridization technique to determine the correlation between CTCs and CTECs and lung cancer disease progression and to identify prognostic indicators. In all patients, the positive rate of CTCs was 100% and the positive rate of CTECs was 81.3%. The CTEC positivity rate was higher in late-stage patients than in early-stage patients (P = .03). Patients with advanced or lymph node metastases had a higher rate of small-size CTC positivity than those with early or no lymph node metastases. Large-size CTEC positivity was higher in patients with advanced NSCLC than in early-stage patients. Patients with ≥1 small-size CTC had shorter progression-free survival, and it was an independent prognostic factor. Small-size CTCs are a reliable prognostic indicator and a probable predictor of the severity of disease in NSCLC patients.

Prognostic Significance of Excision Repair Cross-Complementation Group 1 on Circulating Tumor Cells for Nasopharyngeal Carcinoma.

Liquid biopsy, including the detection of circulating tumor cells (CTCs), has emerged as a promising tool for cancer diagnosis and monitoring. However, the prognostic value of CTCs in nasopharyngeal carcinoma (NPC) remains unclear due to the lack of phenotypic characterization. The expression of Excision Repair Cross-Complementation Group 1 (ERCC1) and CTCs epithelial-mesenchymal transition (EMT) have been associated with treatment efficacy. In this study, we aimed to evaluate the prognostic significance of ERCC1 expression on CTCs and their EMT subtypes before treatment in NPC. We retrospectively analyzed 108 newly diagnosed locally advanced NPC patients who underwent CanPatrol™ CTC testing between November 2018 and November 2021. CTCs were counted and classified into epithelial, epithelial-mesenchymal hybrid, and mesenchymal subtypes. ERCC1 expression was divided into negative and positive groups. Clinical features and survival outcomes were analyzed. The positive rate of CTCs was 92.6% (100/108), with an ERCC1 positivity rate of 74% (74/100). Further analysis of the subtypes showed that positive ERCC1 on mesenchymal CTCs was associated with a later N stage (P = .01). Positive ERCC1 expression was associated with poor overall survival (OS; P = .039) and disease-free survival (DFS; P = .035). Further analysis of subtypes showed that the positive ERCC1 on mesenchymal-type CTCs was associated with poor OS (P = .012) and metastasis-free survival (MFS; P = .001). Our findings suggest that ERCC1 expression on CTCs may serve as a new prognostic marker for NPC patients. Evaluating CTCs subtypes may become an auxiliary tool for personalized and precise treatment.

Detection of circulating tumor cells in patients with pancreatic cancer by novel nano microfluidic chips and comprehensive nursing intervention.

It was to explore the detection efficiency of novel nano microfluidic chips on circulating tumor cells (CTCs) in pancreatic cancer (PC) patients and the effect of comprehensive nursing intervention on the life indicators of patients after PC surgery. PC, benign pancreas, and healthy volunteers were enrolled. The number and positive rate of CTCs in the subjects, the relationship between the number of CTCs in PC patients and clinical data, the recurrence rate, and the life indicators of patients after intervention were sorted out and analyzed. The positive rate of CTCs was 96.67% in PC group (29/30), 16.67% in benign pancreas group (5/30), and 0% in healthy controls. The positive CTCs in PC patients were correlated with vascular invasion (P<0.05). The recurrence rate after PC resection was 56.67% (17/30). The positive detection rate of nano microfluidic chip was 30/30 (100%), which was higher than against CellSeach 7/30 (23.33%), P<0.05. The physiological function score of intervention group (93.67±1.48) was higher as against controls (60.45±2.03), P<0.05. The social function score of intervention group (91.26±0.44) was higher as against controls (54.92±2.98), P<0.05. The novel nano microfluidic chip has high sensitivity and specificity for the detection of CTCs in PC patients. Postoperative comprehensive nursing intervention can improve the quality of life (QoL) and functional status of PC patients.

Identification of Circulating Tumor Cell Phenotype in Differentiated Thyroid Carcinoma

Objective: Circulating tumor cells (CTCs) have been considered as the origin of tumor metastasis and recurrence, which always indicate a poor prognosis. There are three phenotypes of CTCs according on different epithelial-to-mesenchymal transition (EMT) markers, including epithelial, mesenchymal, and epithelial/mesenchymal (mixed phenotypic) CTCs. We intended to explore the relationship among CTC phenotypes and the clinicopathological characteristics of patients with differentiated thyroid carcinoma (DTC). Methods: Peripheral blood samples from 58 patients with DTC were collected, and CTCs were isolated by cell sizes. To identify phenotypes of CTCs, branched DNA signal amplification technology was adopted to capture and amplify target sequences, and then multiplex RNA-in situ hybridization (RNA-ISH) assay was used to identify CTC phenotypes depended on epithelial-mesenchymal transition (EMT) markers. Results: The positive rate of CTCs was 77.59% in 58 DTC patients. Totally, 488 CTCs with detective phenotype were found. Among them, there were 121 (24.80%) epithelial CTCs, 67 (13.72%) mesenchymal CTCs, and 300 (61.48%) mixed phenotypic CTCs. An obvious increased epithelial CTCs was observed in male patients compared with female. Notably, CTCs were more prevailing in younger male patients with ETI and bilateral focus. Conclusions: The CTCs are common in DTC patients, and mixed phenotypic is the major phenotype, indicating that EMT is prevalent in DTC even though its prognosis was better than other epithelial tumors. Detection of CTC and its phenotypes might independently predict the prognosis of DTC.

Correlation between circulating tumor cells and different molecular biological characteristics in breast cancer patients.

This study sought to detect the number of circulating tumor cells (CTCs) in breast cancer patients, and examine the relationship between CTCs and molecular biological characteristics. From June 2016 to June 2018, 150 female patients with invasive breast cancer detected by CTCs at the Department of Breast Surgery, Fudan University Cancer Hospital were enrolled in this study. The patients had an average age of 52.6±7.8 (range, 35-77) years. Routine pathological and immunohistochemical examinations were performed on tissues obtained during surgery. In this study, CTCs were detected using the immunomagnetic bead negative enrichment technique (i.e., the Cyttel technique). The measurement data are expressed by x ± s, and were compared by t-tests. A univariate analysis of variance was used to compare differences between groups. The count data are expressed as the absolute value, and the test χ2 or Fisher's exact test were used to compare differences. There were 109 cases of positive CTC (≥3 CTCs/4 mL) (72.7%), and 41 cases of negative CTC (<3 CTCs/4 mL) (27.3%). There were no significant differences in terms of age and menopausal status between the two groups (P>0.05). There was no significant difference in the positive rate of CTC in T1, T2, and T3 and above patients (P>0.05). There was no significant difference in the CTC positive rate between ER positive and negative patients, PR positive and negative patients, and Ki-67 ≥14% and <14% patients (P>0.05). However, there was a statistical difference in the positive rate of CTC between human epidermal growth factor receptor 2 (HER-2) positive and negative patients (P<0.05). There was no significant difference in the CTC positive rate among patients with Luminal A type, B type, HER-2 overexpression type, and triple negative breast cancer (TNBC) (P>0.05). There was no significant difference in the positive rate of CTC among patients with invasive ductal carcinoma, invasive lobular carcinoma, and other types of invasive carcinoma (P>0.05). It can be concluded that there is a relationship between CTC and HER-2 expression, which has certain predictive value for patients with positive HER-2 expression, thus predicting poor prognosis.

The clinical significance of circulating tumor cells and T lymphocyte subtypes in pancreatic cancer patients

ABSTRACT Circulating tumor cells (CTCs) are sensitive and reliable biomarkers for tracing relapsed and metastatic cancer. Here, we explore the clinical significance of CTCs and T lymphocyte subtypes in patients with pancreatic cancer. A total of 106 patients with the pancreatic cancer were enrolled in this study. The enrichment and identification of CTCs were achieved before treatment by a PatrolCTC detection technique. Flow cytometry (FACS) was used to characterize CD4, CD8, natural killer (NK) cells, and Tregulatory (Treg) lymphocyte subtypes. Interleukin-2 (IL-2), Interleukin-4 (IL-4), Interleukin-17A (IL-17A), Interleukin-10 (IL-10), and Interferon γ (IFN-γ) were measured by meso-scale discovery (MSD) assay. Among these patients, 44 (41.5%) patients with pancreatic ductal adenocarcinoma (PDAC) were female and 62 (58.5%) cases were male. Case numbers with II–IV tumor-node-metastasis (TNM) stages were 32 (30.2%), 50 (47.2%), and 24 (22.6%), respectively. The positive rate of CTCs before surgery was 37.5% (12/32), 88.0% (44/50) and 100% (24/24) in stage II, III, and IV patients, respectively. Total CTCs, mixed CTCs, and mesenchymal CTCs (MCTCs) were strongly relevant to shorter progression-free survival (PFS) of the patients. In addition, total CTCs (≥6) and positive MCTCs were also significantly correlated with recurrence and metastasis. The patients with high CTCs also had low levels of CD4, CD4/CD8 ratio, NK cells, IL-2, and IFNγ. In contrast, Treg cells had significant elevation in PDAC patients. These results indicated that CTCs number in PDAC patients was an independent indicator for worse PFS. High CTCs also had strong correlation with weak cellular immunity functions.

The Correlations among Circulating Tumor Cells, Epstein-Barr Virus Status, and Epidemiology in Patients with Nasopharyngeal Carcinoma.

Studies have shown that circulating tumor cells (CTCs) can be detected in nasopharyngeal carcinoma (NPC). However, the relationship between CTCs and tumor stage is still controversial. This study aims to investigate the correlations among CTCs, Epstein-Barr virus (EBV) status, clinicopathologic features, and epidemiological risk factors in patients with NPC. Three hundred and thirty primary NPC patients with complete clinical data and epidemiology information were collected. Analysis of CTCs was performed using the CTCBIOPSY system. The plasma EBV DNA load was detected by quantitative real-time PCR. Detection of VCA-IgA and EA-IgA antibodies titers was conducted by immunoenzymatic assay. EBNA1-IgA and Zta-IgA were measured using an enzyme-linked immunosorbent assay. The presence of CTCs was associated with high EBV DNA load (p < 0.05). The positive rate of CTCs was correlated with T and M classifications of NPC (T: 13.2% vs. 22.9%; M: 17.9% vs. 34.8%, p < 0.05). Compared with never and former smokers, current smokers exhibited a higher positive rate of EBNA1-IgA (83.3% and 81.0% vs. 92.5%, p < 0.05); the patients with pack-years of smoking ≥ 15 displayed a significantly higher positive rate of EBNA1-IgA than those with pack-years of smoking < 15 (98.0% and 92.5% vs. 81.0%, p < 0.05). CTCs positivity was closely associated with tumor burden and distant metastasis of NPC. Smoking status and smoking cumulative dose of NPC patients might be correlated with EBV activation.

Optimal Strategy for Colorectal Cancer Patients' Diagnosis Based on Circulating Tumor Cells and Circulating Tumor Endothelial Cells by Subtraction Enrichment and Immunostaining-Fluorescence In Situ Hybridization Combining with CEA and CA19-9.

Background Cancerous embryo antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) are commonly used in clinical practice to assist in diagnosing CRC. However, their sensitivity is very low. This study aims to investigate the clinical significance of circulating tumor cells (CTCs) and circulating tumor endothelial cells (CTECs) compared with CEA and CA19-9 in the auxiliary diagnosis of colorectal cancer (CRC) patients. Methods 115 pathologically confirmed CRC patients and 20 healthy controls were enrolled in this study. CTCs and CTECs were enriched and identified by subtraction enrichment and immunostaining-fluorescence in situ hybridization (SE-iFISH). A logistic regression was used to establish a model for the receiver-operating characteristic (ROC) curve analysis, and the diagnostic efficacy of CTCs, CTECs, CEA, CA19-9, and their combinations was analyzed. Results The CTC (P < 0.0001) and CTEC (P=0.0009) level was significantly higher in CRC patients than that in healthy controls. For CRC patients, CTC and CTEC level was significantly correlated with tumor stage and lymph node metastasis status, but not with sex, age, tumor location, and degree of differentiation. The positive rate of CTCs, CTECs, CEA, and CA19-9 in CRC patients was 87.8%, 39.1%, 28.7%, and 26.1%, respectively. To distinguish CRC patients from controls, the area under the curve (AUC) of CTC was 0.889, which was much higher than 0.695 of CTEC, 0.696 of CEA, and 0.695 of CA19-9. Establishing ROC curve by logistic regression algorithm, the highest AUC was 0.935, which combined CTCs with CTEC, CEA, and CA19-9. Conclusions CTCs combined with CTEC, CEA, and CA19-9 are useful to improve the diagnostic efficiency, which has high clinical significance in the diagnosis of colorectal cancer.

Simultaneous in situ detection of protein expression of multiple tumor markers of circulating tumor cells and heteroploid of chromosome 8 in primary lung cancer

BackgroundPreviously published studies have shown that circulating tumor cells (CTC) play an important role in clinical staging, efficacy monitoring and prognostic evaluation of lung cancer. The aim of the present study was to investigate the significance of simultaneous in situ detection of multiple tumor marker protein expression and chromosome 8 aneuploid of CTC in the diagnosis and treatment of primary lung cancer.MethodsWe investigated the expression of cytokeratin 18 (CK18) and Vimentin on the surface of CTC and the aneuploidy of chromosome 8 in the peripheral blood of 24 patients with metastatic primary lung cancer, which were detected by subtraction enrichment (SE) and immunofluorescence in situ hybridization (iFISH). Their correlation with clinicopathological features, curative effect, and prognosis was analyzed.ResultsThe positive rate of CTC was 95.83% (23/24). There was a certain correlation between the positive rate of CTC and smoking, and a correlation between the number of CTC and histopathological type. The number of monoploid, diploid, triploid, and polyploid CTC had no statistical correlation with progression-free survival (PFS) or overall survival (OS). However, a tetraploid CTC count of ≥2 was an unfavorable predictor of response; a tetraploid CTC count ≥1 was an unfavorable prognostic predictor of poor OS. The rate of CK18+ CTC in all 24 patients was 4.35% (1/23). Seven out of these 24 patients were also tested for Vimentin, and the rate of Vimentin+ CTC was 85.71% (6/7). Small-cell (≤5 µm) CTC was found in 6 of these 7 patients and accounted for 11.83% (11/93) of the total CTC. The tumor marker phenotype of small-cell CTC was CK18– and Vimentin+. In addition, circulating tumor microthrombi (CTM) were found in 2 of these 7 patients (28.57%).ConclusionsSE-iFISH has a high detection rate of CTC in the peripheral blood of patients with metastatic primary lung cancer, and it can identify small cell CTC. Tetraploid CTC count ≥2 can predict poor PFS in patients with advanced lung cancer. Tetraploid CTC count ≥1 could predict poor OS in patients with advanced lung cancer. CTM can predict poor prognosis in patients with lung cancer.

Perioperative Circulating Tumor Cells (CTCs), MCTCs, and CTC-White Blood Cells Detected by a Size-Based Platform Predict Prognosis in Renal Cell Carcinoma.

To explore the clinical significance of the perioperative counts of circulating tumor cells (CTCs), mesenchymal CTCs (MCTCs), and CTC- white blood cells (WBCs) in renal cell carcinoma patients. A total of 131 patients with renal cancer who underwent operation excision from our hospital were enrolled. In addition, 20 patients with benign renal diseases were recruited as a control. Blood samples were collected from the 131 patients, before operation and 3 months after surgery. Samples were also obtained simultaneously from the control group. CanPatrol CTC detection technique was used to enrich and identify CTCs, MCTCs, and CTC-WBCs. All enrolled patients were T1-3N0M0. From these, 52 patients with renal cancer underwent radical resection, while other 79 patients underwent nephron-sparing surgery. The positive rate of CTC, MCTC, and CTC-WBC before surgery were 95.4% (125/131), 61.1% (80/131), and 11.5% (15/131), respectively. Preoperative total CTCs, MCTCs, or CTC-WBCs were poorly correlated with patients' parameters. Preoperative CTC, MCTC, or CTC-WBC showed no association with progression-free survival (PFS). In contrast, postoperative total CTCs (≥6), positive MCTCs, and positive CTC-WBCs significantly correlated with recurrence and metastasis. These results remained independent indicators for worse PFS. In addition, the increased CTC and MCTC count after surgery also correlated with unfavorable PFS. The detection of six or more total CTCs, MCTC, or CTC-WBCs in peripheral blood after surgery might help to identify a subset of patients that have higher recurrent risk than the overall population of patients with at different stages of renal cancer.

Detection of circulating tumor cells with chromosomes 7 and 8 polysomy in non-small cell lung cancer and its correlation with epidermal growth factor receptor mutations in cancer tissue

Objective: To investigate the value of chromosomes 7 and 8 polysomy in circulating tumor cells (CTCs) for the diagnosis of non-small cell lung cancer, and the correlation of CTCs with clinical pathological characteristics and epidermal growth factor receptor (EGFR) mutations in cancer tissue. Methods: Fifty-seven patients with non-small cell lung cancer and 21 patients with benign lung diseases were enrolled at Beijing Chaoyang Hospital, Capital Medical University, Beijing, China from November 2017 to October 2020. Negative enrichment combined with immunofluorescence in situ hybridization (imFISH) was used to identify CTCs polysomy on chromosomes 7 and 8. EGFR mutations in 56 lung cancer patients was detected using ARMS-PCR. Results: CTCs were detected in 93.0% (53/57) of non-small cell lung cancers and 28.6% (6/21) benign lung lesions. The difference between lung cancer patients and the control cohort was statistically significant (P<0.01). Receive operator curve (ROC) analyses showed that, when the cut-off value was 1 cell/3.2 mL, Youden index had the highest sensitivity of 93.0% and specificity of 71.4% (AUC=0.906, 95%CI:0.833-0.980, P<0.01). The positive rate of CTCs in stage Ⅲ-Ⅳ cancers was significantly higher than that in stage Ⅰ-Ⅱ (P=0.023). No significant correlation was observed between positive rate of CTCs or chromosome polysomy and age, gender, smoking status, pathologic types and EGFR mutation status. The number of CTCs in EGFR mutated group was higher than that in the non-mutated group (6.5±1.1 vs. 3.7±0.7, P=0.045). The detection rate for CTCs ≥5 in the EGFR mutated group was also higher than the EGFR non-mutated group (52.0% vs. 19.4%,P=0.010). Conclusion: Detection of CTCs with chromosomes 7 and 8 polysomy has potential value in auxiliary diagnosis of non-small cell lung cancer, and the number of CTCs is correlated to TNM stage and EGFR gene mutation status.

Utility of circulating tumor cells in stage II colorectal cancer patients undergoing curative resection.

BackgroundThe aim of this study was to determine whether circulating tumor cells (CTCs) have utility as a prognostic biomarker in stage II colorectal cancer (CRC), as well as a biomarker for the selection of patients for adjuvant chemotherapy.MethodsCTCs were detected in peripheral blood samples obtained from 73 stage II CRC patients, using a negative enrichment and immune-fluorescence in situ hybridization (imFISH) staining method. The follow-up time ranged from 3.5 to 35.9 months, and the clinic-pathologic characteristics and recurrence free survival (RFS) were collected and analyzed.ResultsSeventy-three stage II CRC patients were included in this study. The positive rate of CTCs was 65.8% in all patients, 87.5% in recurrent patients and 59.6% in no recurrence patients. The mean RFS was 30.6 months for all patients, 28.7 months for CTC-positive patients and 34.0 months for CTC-negative patients (P=0.043). The mean RFS of CTC-positive and CTC-negative patients with adjuvant chemotherapy were not reached, and those without adjuvant chemotherapy were 27.7 and 33.4 months, respectively.ConclusionsThe level of CTCs may be an effective prognostic factor to predict RFS in stage II CRC patients, and has potential in selecting stage II CRC patients for adjuvant chemotherapy.

Circulating tumor cells in whole process management of gastrointestinal stromal tumor in a real-life setting.

Background/Aim:Liquid biopsy is changing the diagnosis and treatment strategies of various neoplasms. However, the circulating tumor cells (CTCs) of gastrointestinal stromal tumor (GIST) patients with different disease process are not clear. To better understand the dynamic change of CTCs in GIST patients, we conducted a real-life setting study.Patients and Methods:One-hundred fifty GIST patients were included. The isolation by size of tumor cell (ISET) method was employed to detect the CTCs/circulating tumor microemboli (CTM). Imatinib (IM) plasma concentration was detected by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Multivariate and univariate analysis were used to analyze the effects of clinical characteristics on the positive rate of CTC and the number of CTCs/CTM.Results:The positive rate of CTCs was 72%. The median number of CTCs and CTM was 4 and 0. Logistic multivariate regression analysis showed that tumor diameter was the only independent factor of the positive rate of CTCs (P < 0.05). The numbers of CTCs and CTM had intensive linear correlation (P < 0.001). Tumor diameter, Ki 67 expression and mitotic were related to the number of CTCs (P < 0.05). Patients with higher Ki 67 expression tend to have more CTM (P < 0.05). IM plasma concentration showed no influence to the CTCs/CTM (P > 0.05).Conclusions:In the current study, we assessed the CTCs and CTM of GIST patients in various disease progressions and identified clinicopathological factors influencing the detection of CTCs and CTM. These results are instructive for clinicians to understand CTCs/CTM in GIST patients.

Correlation between circulating tumor cells and clinicopathological features of early breast cancer

To investigate the correlation between the number of peripheral blood circulating tumor cells (CTCs) and clinicopathological features of early breast cancer. Methods: The clinical and pathological data from 100 patients with early breast cancer treated by a breast cancer treatment team in the Department of Breast Surgery, Second Xiangya Hospital, Central South University, were collected from January 2017 to December 2018. For these patients, their peripheral blood CTCs were detected, enumerated and typed by CanpatrolTM CTC assay. Results: The positive rate of CTCs was 90% in peripheral blood of patients with early breast cancer, and the majority of molecular phenotypes was hybrid CTCs (55.6%). The positive rate of CTCs was only related to the pathological type of tumor (P<0.05), but not to other clinicopathological features. No correlation between clinicopathological features and the total number of CTCs, the number of epithelial CTCs or the number of hybrid CTCs was found. However, the number of mesenchymal CTCs was significantly correlated with the expression of hormone receptors and Ki-67 (r=0.200, P<0.05), and there was a significant correlation between the proportion of mesenchymal CTCs and the expression level of Ki-67 (r=0.213, P<0.05). Conclusion: The number of CTCs is not correlated with all clinicopathological features, but patients with negative hormone receptor and high expression of Ki-67 probably have more hybrid CTCs.

Significance of postoperatively detecting peripheral blood circulating tumor cells in patients with papillary thyroid cancer

Objective To investigate the significance of postoperatively detecting peripheral blood circulating tumor cells (CTCs) in the patients with papillary thyroid cancer (PTC). Methods A total of 143 patients diagnosed as having PTC in our department, receiving radical thyroidectomy were analyzed. The CTCBIOPSY D-100 equipment developed by You Zhiyou Medical Technology Co., Ltd. as well as ISET separation technique, were used to count the number of peripheral blood CTCs, and its relationship with the clinical features and prognosis of the patients were analyzed by the statistical method. Results The positive rate of CTCs in all 143 patients with PTC was 58.74%. Statistical analysis showed that the positive rate of peripheral blood CTCs in PTC patients was significantly correlated with TNM stage, tumor’s invasion of muscle and/or recurrent laryngeal nerve, and pathological lymphatic metastasis (P 0.05). Logistic regression statistical analysis indicated that the factors indicating poor prognosis of PTC such as TNM stage, tumor’s invasion of muscle and/or recurrent laryngeal nerve, lymphatic metastasis were independent influencing factors of CTCs [odds ratio (OR): 2.417, 3.168, 3.101, P<0.05]. Conclusion Positive CTCs in the peripheral blood postoperation may indicate a poor prognosis in patients with PTC. Key words: Papillary thyroid cancer; Circulating tumor cells; Tumor recurrence

Isolation of Circulating Tumor Cells of Ovarian Cancer by Transferrin Immunolipid Magnetic Spheres and Its Preliminary Clinical Application

Circulating tumor cells (CTCs) play an important role in cancer prognosis, treatment monitoring and metastasis diagnosis. However, due to the extremely low concentration of CTC in the peripheral blood, its isolation and enrichment are critical steps for early diagnosis. Herein, we used the transferrin modified lipid magnetic spheres for the isolation of ovarian cancer CTCs, and studied the relationship between the CTCs count and the clinical case parameters, prognosis of ovarian cancer. The result showed that no CTC was found in the peripheral blood of 30 patients with benign cysts, and 34 out of 46 patients with ovarian cancer were positive for CTC, with a positive rate of 73.9%. Analysis of the parameters of the clinical cases showed that the positive rate of CTC was related to the clinical stages, and that it was not significantly related to the age, histopathological types and pathological grades of patients. Of the 34 CTC-positive patients, 18 had progression-free survival, with a survival rate of 52.9%, and of the 11 CTC-negative patients, 9 had progression-free survival, with a survival rate of 81.8%. The results showed that the transferrin lipid magnetic spheres prepared in this study, could effectively isolate the CTCs in the peripheral blood of patients with ovarian cancer, that the level of CTC in ovarian cancer patients was related to its clinical stage, and that the progression-free survival of the patients with a high level of CTCs was relatively short. Therefore, this study shows that the transferrin lipid magnetic sphere can achieve effective isolation of ovarian cancer CTC, which can be used as an auxiliary diagnostic method in comprehensive diagnosis of ovarian cancer.

The value of circulating tumor cells detected by chromosome centromere probe identification in diagnosis of non-small cell lung cancer

Objective: This study explored the value of circulating tumor cells (CTC) detected by chromosome 8 centromere probe in diagnosis of non-small cell lung cancer (NSCLC) as well as correlation between CTC counts and clinical pathological characteristics. Methods: We collected 136 patients with newly diagnosed NSCLC (101 males and 35 females, age ranged from 34 to 79 years), 149 patients with non-malignant pulmonary diseases (103 males and 46 females, age ranged from 24 to 80 years) and 32 healthy volunteers (5 males and 27 females, age ranged from 24 to 42 years). Detection was performed using an epithelial cell adhesion molecule-independent strategy that combined immunocytochemistry staining (ICC) of CD(45) and fluorescence in situ hybridization detection (FISH) with chromosome 8 centromere probe (CEP8). CTC was defined as 4',6-diamidino-2-phenylindole (DAPI) positive, CD(45) negative and CEP8 more than 2 positive points. Quantitative data were reported as x±s and Mann Whitney test was used to compare them. Measurement data were analyzed as contingency tables and Pearson chi-squared test was used. P values of less than 0.05 were considered statistically significant. Results: CTCs were detected in 114 patients (83.8%) with NSCLC, 35 patients (23.5%) with non-malignant pulmonary diseases (P<0.000 1) and 5 volunteers (15.6%). CTC counts in NSCLC patients were (5.98±0.64) per 3.2 ml and (0.60±0.13) per 3.2 ml (P<0.000 1). A receiver operating characteristic curve (ROC) analysis showed similar capability of CTC count, with CEA, in discriminating NSCLC and non-malignant diseases with an area under ROC curve of 0.854 (95% confidence interval: 0.808-0.900, P<0.001). Cutoff of 2 circulating tumor cells per 3.2 ml peripheral blood gave the highest Youden Index of 0.614 in diagnosis of NSCLC with sensitivity of 72.1% (98/136) and specificity of 89.3% (133/149). When patients with CTC≥2/3.2 ml peripheral blood or serum CEA≥5 ng/ml were considered as having NSCLC, sensitivity and specificity of this combined test were 87.5% (119/136) and 85.9% (128/149), with a higher Youden Index of 0.734. No correlation was observed between positive rate of CTC (rate of patients with CTC≥2/3.2 ml peripheral blood) and age, gender, smoking status, pathologic types, and clinical stages. Conclusions: CTC counts detected by CD(45)-FISH is higher in NSCLC patients than in non-malignant disease patients. CTC≥2/3.2 ml peripheral blood is valuable in discriminating NSCLC from nonmalignant diseases, which can be more accurate when combined with CEA.

Abstract 1577: Correlation of exosomes and CTCs in non-small cell lung cancer patients with disease stage

Abstract Lung cancer is the most common cancer, which also leading cause of global cancer mortality. Metastasis is a complex multi-step process that represents the most deadly aspect of cancer. One key step in metastasis is the entry of circulating tumor cells (CTCs) into secondary or distant organ sites to become disseminated tumor cells for subsequent metastasis. Exosomes, a subset of extracellular vesicles (EVs), function as a mode of intercellular communication and molecular transfer. Tumor cells are avid producers of exosomes, and tumour-derived exosomes uptaken by organ-specific cells prepare the pre-metastatic niche. The aim of this pilot study is to investigate the relationship between the count of circulating tumor cells in peripheral blood and the clinical stage of lung cancer, and the difference between normal (healthy) person exosomes and cancer patients exosomes in the rate of exosomes release and the content (most notably proteins). A total of 140 patients participated in the project and twelve blood samples from healthy volunteers were also collected in the Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. After obtaining the informed consent, blood samples (5 ml) were stored in the ACD anticoagulant tube. CTCs was counted by immunomagnetic beads negative enrichment technique combined with immunofluorescence staining of chromosome in situ hybridization. The correlation between the count of CTCs and the clinical stage was analyzed. Exosomes were isolated by Density Gradient (DG) centrifugation. Exosomes concentration and size was analyzed by Nanoparticle Tracking Analysis (NTA), and protein was analyzed by HPLC-CHIP-MS/MS. In 114 patients, the positive rate of CTCs was 93.86% (in 3.2mL blood CTCs≥1, 107/114). The count of CTCs was negatively correlated with clinical stage (rs=-0.279, p=0.003), which suggest that CTCs is highly expressed in early lung cancer patients. Exosomes test results showed that there was a significant difference between lung cancer patients and healthy person. The size of exosome in healthy person was significantly smaller and the concentration was significantly higher when compared with lung cancer patients (p&amp;lt;0.05). In addition, the mass spectrometry results showed that the control group had 135 proteins in the exosomes, while the patient group contained 167 proteins, which 89 proteins were shared with each other. These results suggest that CTCs plays an important role in the pathogenesis of lung cancer and should be a target for early postoperative metastasis. The analysis of exosomes in NSCLC could represent a noninvasive test for assess disease progression. Citation Format: Zujun Que, Yi Jiang, Fangfang Qian, Zhi Qiao, Hua Xiao, Lingshuang Liu, Hegen Li, Jianhui Tian. Correlation of exosomes and CTCs in non-small cell lung cancer patients with disease stage [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 1577.

Relationships between peripheral circulating tumor cells in patients with gallbladder carcinoma detected by a novel nano microfluidic chip with clinicopathology and prognosis

Objective To establish a novel method using a nano microfluidic chip to detect circulating tumor cells (CTCs) in peripheral blood in gallbladder carcinoma, and to study the relationship between CTCs with clinicopathology and prognosis in these patients. Methods The peripheral blood samples of 51 patients with gallbladder carcinoma were collected from June 2014 to January 2017. The CTCs from peripheral blood samples were detected by a novel nano microfluidic chip. This study aimed to study the correlation between CTCs with the clinical and pathological features. The significance of CTCs on prognosis in patients with gallbladder carcinoma was also analyzed. Results The positive rate of CTCs in the peripheral blood of gallbladder carcinoma patients was 43.1% (22/51). There were significant correlations between CTCs with liver metastasis (P 0.05). Conclusions Peripheral blood CTCs in patients with gallbladder carcinoma were efficaciously detected by a novel nano microfluidic chip. Peripheral blood CTCs was closely related to the Nevin staging and liver metastasis. CTCs could serve as a potential prognostic indicator in patients with gallbladder carcinoma. Key words: Gallbladder carcinoma; Microfluidic chip, nano; Circulating tumor cells (CTCs); Prognosis

Correlation between sodium-iodide symporter expression and circulating tumor cell positivity in differentiated thyroid carcinoma*

Abstract Objective We investigated the correlation between the expression of the sodium-iodide symporter (NIS) and the detection of circulating tumor cells (CTCs) in differentiated thyroid carcinoma (DTC). Methods NIS expression in differentiated thyroid and the positive rate of CTCs in the peripheral blood were determined by immunohistochemistry S-P and flow cytometry from the records of 172 cases of differentiated thyroid carcinoma. Results Seventy-six cases (44.2%) expressed NIS in the differentiated thyroid and 63 cases (36.6%) were positive for CTCs in the peripheral blood. There was a significant difference between N0 and N1 in the expression of NIS (χ2 = 6.015, P = 0.014) and the positive rate of CTCs (χ2 = 14.035, P = 0.001). N0 and N1 also differed significantly in the expression of NIS (r = -0.383, -0.610, P = 0.002, &lt; 0.001). The differences in the NIS expression, but not in the positive rate of CTCs, were significant among the different pathological subtypes (χ2 = 7.897, P = 0.005; χ2 = 1.455, P = 0.228, respectively). There was a significant negative correlation between the highly differentiated type and intermediate differentiation type both in the expression of NIS and positive rate of CTCs (r = -0.591, -0.443, P &lt; 0.001, P = 0.002). Conclusion There was a significant negative correlation between the expression of tissue NIS and positive rate of CTCs in the peripheral blood in DTC. The malignancy level and lymph node metastasis in differentiated thyroid carcinoma were negatively correlated with NIS expression and positively correlated with the positive rate of CTC.