Improvement of sensitive and specific detection of circulating tumor cells using negative enrichment and immunostaining-FISH

Li, Yang; Ma, Guojun; Zhao, Peige; Fu, Rao; Gao, Lei; Jiang, Xiaohong; Hu, Ping; Ren, Tianying; Wu, Yaping; Wang, Zhongye; Cui, Zhaoqing; Yang, Dawei

doi:10.1016/j.cca.2018.06.034

Cited by 31 publications

(34 citation statements)

References 44 publications

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“…The present study aimed to explore the role of EMT in the immune escape of gastric CTCs. We used Cyttel‐CTC and im‐FISH (immune‐fluorescence in situ hybridization) methods to detect CTCs and to investigate the expression of EMT markers and ULBP1 on CTCs . Moreover, we investigated the relationship between the expression of EMT markers and ULBP1 in vitro in gastric cancer cell lines.…”

Section: Introductionmentioning

confidence: 99%

Epithelial‐mesenchymal transition may be involved in the immune evasion of circulating gastric tumor cells via downregulation of ULBP1

Tian

et al. 2020

Cancer Medicine

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Background:Increasing numbers of studies have demonstrated that circulating tumor cells (CTCs) undergo a phenotypic change termed epithelial-mesenchymal transition (EMT), and researchers have proposed that EMT might provide CTCs with increased potential to survive in the different microenvironments encountered during metastasis through various ways, such as by increasing cell survival and early colonization. However, the exact role of EMT in CTCs remains unclear. Methods: In this study, we identified CTCs of 41 patients with gastric cancer using Cyttel-CTC and im-FISH (immune-fluorescence in situ hybridization) methods, and tested the expression of EMT markers and ULBP1 (a major member of the NKG2Dnatural killer [NK] group 2 member D-ligand family) on CTCs. Moreover, we investigated the relationship between the expression of EMT markers and ULBP1 on CTCs and gastric cancer cell lines. Results: Our results showed that the CTCs of gastric cancer patients exhibited three EMT marker subtypes, and that the expression of ULBP1 was significantly lower on mesenchymal phenotypic CTCs (M + CTCs) than on epithelial phenotypic CTCs (E + CTCs). EMT induced by TGF-β in vitro produced a similar phenomenon, and we therefore proposed that EMT might be involved in the immune evasion of CTCs from NK cells by altering the expression of ULBP1. Conclusions: Our study indicated that EMT might play a vital role in the immune invasion of CTCs by regulating the expression of ULBP1 on CTCs. These findings could provide potential strategies for targeting the immune evasion capacity of CTCs.

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Section: Introductionmentioning

confidence: 99%

Epithelial‐mesenchymal transition may be involved in the immune evasion of circulating gastric tumor cells via downregulation of ULBP1

Tian

et al. 2020

Cancer Medicine

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“…In this study, an EpCAM‐independent enrichment strategy and FISH were used to detect CTCs in lung cancer patients . The relationship between CTC numbers, the levels of particular tumor markers (CEA, CA 125, CYFRA 21‐1, and SCC), and the clinicopathological factors in these patients were also analyzed.…”

Section: Introductionmentioning

confidence: 99%

“…9,[20][21][22][23][24][25][26] In this study, an EpCAM-independent enrichment strategy and FISH were used to detect CTCs in lung cancer patients. 27 The relationship between CTC numbers, the levels of particular tumor markers (CEA, CA 125, CYFRA 21-1, and SCC), and the clinicopathological factors in these patients were also analyzed. Moreover, the potential clinical use of the combination of CTCs and tumor markers for the diagnosis of lung cancer was explored as well.…”

Section: Introductionmentioning

confidence: 99%

Clinical significance of circulating tumor cells and tumor markers in the diagnosis of lung cancer

et al. 2019

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Background Lung cancer has the highest fatality rate of all cancer types. To improve patients’ survival and life quality, it is therefore very important to screen for and detect it at an early stage. Methods A negative enrichment–fluorescence in situ hybridization (NE‐FISH) approach was used to detect circulating tumor cells (CTCs) in lung cancer patients, and levels of lung cancer‐associated serum markers were also measured in the peripheral blood of these same patients. The correlation between CTCs, serum cancer markers (carcinoembryonic antigen [CEA], CA 125, CYFRA 21‐1, and SCC), and clinicopathological characteristics was then investigated. Moreover, the potential clinical use of the combination of CTCs and tumor markers for the diagnosis of lung cancer, especially at early stages, was also explored. Results CTC frequencies in lung cancer patients were significantly higher than in healthy control volunteers or patients with benign lung disease, and the area under the receiver operating characteristics curve for the control group was 0.846 (95% CI 0.796‐0.887, P < 0.001). The rate of CTC positivity in lung cancer patients was 68.29% when the CTC cutoff value was 2, and the sensitivity of this means of lung cancer detection rose to 82.93% by combining CTC‐based detection with measurements of serum tumor markers. Similarly, the diagnostic sensitivity of this approach in early‐stage lung cancer patients (I‐II) was improved from 63.93% to 78.69%. Detection of CTCs can thus assist with the identification of benign and malignant pulmonary nodules. Conclusions It is potentially helpful and effective to employ a combination of CTCs and serum tumor markers for the clinical diagnosis of lung cancer.

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“…Most CellSearch studies set the standard at 5 CTCs/7.5 mL for BC ever since Cristofanilli's group demonstrated that the baseline level of ≥ 5 CTCs/7.5 mL was associated with shorter PFS and OS [40,41]. Reported standards for other methods include ≥ 1, ≥ 2, and ≥ 3 CTCs in 5-7.5 mL of blood sample [20,40,42]. It is reasonable to define different positive criteria of CTCs for diverse methods since the compositions of captured CTCs are not the same depending on the enrichment principle.…”

Section: Discussionmentioning

confidence: 99%

Metabolic classification of circulating tumor cells as a biomarker for metastasis and prognosis in breast cancer

Chen

Dong

et al. 2020

J Transl Med

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Background: Circulating tumor cells (CTCs) has been demonstrated as a promising liquid biopsy marker for breast cancer (BC). However, the intra-patient heterogeneity of CTCs remains a challenge to clinical application. We aim at profiling aggressive CTCs subpopulation in BC utilizing the distinctive metabolic reprogramming which is a hallmark of metastatic tumor cells. Methods: Oncomine, TCGA and Kaplan-Meier plotter databases were utilized to analyze expression and survival relevance of the previously screened metastasis-promoting metabolic markers (PGK1/G6PD) in BC patients. CTCs detection and metabolic classification were performed through micro-filtration and multiple RNA in situ hybridization using CD45 and PGK1/G6PD probes. Blood samples were collected from 64 BC patients before treatment for CTCs analysis. Patient characteristics were recorded to evaluate clinical applications of CTCs metabolic subtypes, as well as morphological EMT subtypes classified by epithelial (EpCAM/CKs) and mesenchymal (Vimentin/Twist) markers. Results: PGK1 and G6PD expressions were up-regulated in invasive BC tissues compared with normal mammary tissues. Increased tissue expressions of PGK1 or G6PD indicated shortened overall and relapse-free survival of BC patients (P < 0.001). Blood GM + CTCs (DAPI + CD45 − PGK1/G6PD +) was detectable (range 0-54 cells/5 mL) in 61.8% of tCTCs > 0 patients. Increased GM + CTCs number and positive rate were correlated with tumor metastasis and progression (P < 0.05). The GM + CTCs ≥ 2/5 mL level presented superior AUC of ROC at 0.854 (95% CI 0.741-0.968) in the diagnosis of BC metastasis (sensitivity/specificity: 66.7%/91.3%), compared with that of tCTCs (0.779) and CTCs-EMT subtypes (E-CTCs 0.645, H-CTCs 0.727 and M-CTCs 0.697). Moreover, GM + CTCs + group had inferior survival with decreased 2 years-PFS proportion (18.5%) than GM + CTCs − group (87.9%; P = 0.001). Conclusions: This work establishes a PGK1/G6PD-based method for CTCs metabolic classification to identify the aggressive CTCs subpopulation. Metabolically active GM + CTCs subtype is suggested a favorable biomarker of distant metastasis and prognosis in BC patients.

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Improvement of sensitive and specific detection of circulating tumor cells using negative enrichment and immunostaining-FISH

Cited by 31 publications

References 44 publications

Epithelial‐mesenchymal transition may be involved in the immune evasion of circulating gastric tumor cells via downregulation of ULBP1

Epithelial‐mesenchymal transition may be involved in the immune evasion of circulating gastric tumor cells via downregulation of ULBP1

Clinical significance of circulating tumor cells and tumor markers in the diagnosis of lung cancer

Metabolic classification of circulating tumor cells as a biomarker for metastasis and prognosis in breast cancer

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