Current progress of liquid biopsy in early diagnosis of hepatocellular carcinoma / 中华肝胆外科杂志

Early diagnosis of hepatocellular carcinoma (HCC) is of great significance in the management of patients. Liquid biopsy is a promising tool to use for early diagnosis of liver cancer by detecting tumor expressions through analyzing circulating tumor components such as circulating tumor DNA, circulating tumor cells and extracellular vesicles. The advantages of using liquid biopsy include easy collection of specimen samples and its good sensitivity and specificity for HCC detection. In this review, recent research progress on liquid biopsy on HCC is discussed with the aim to provide updated information on early diagnosis of HCC.

Clinical value of circulating tumor cell typing and RAS/RAF gene detection in colorectal cancer / 中国医师杂志

Objective:To investigate the correlation between circulating tumor cell (CTC) detection, RAS/RAF gene mutation and clinicopathological characteristics in patients with colorectal cancer (CRC).Methods:The Amplification Refractory Mutation System (ARMS)-polymerase chain reaction (PCR) were used to detect the gene mutation in the tumor tissues of 138 CRC patients in the Third Affiliated Hospital of Sun Yat-sen University from May 2017 to May 2020. At the same time, the venous blood of 138 patients was collected and enriched for CTC genotyping by mRNA in situ hybridization. The correlation between CTC, RAS/RAF gene mutation and clinicopathological features of CRC patients was analyzed.Results:The mutation rates of KRAS, NRAS and BRAF genes were 48.6%(67/138), 5.1%(7/138) and 1.4%(2/138), respectively; The overall positive rate of CTC was 84.1%(116/138). The positive rates of different CTC types were: 23.1%(32/138) in epithelial type, 71.7%(99/138) in mixed type and 12.3%(17/138) in interstitial type respectively. The positive rate of CTC in CRC patients with clinical stage Ⅲ-Ⅳ, lymph node metastasis (N1-N3) and distant metastasis (M1) was significantly higher than that in CRC patients with stage Ⅰ-Ⅱ, no lymph node metastasis (N0) and no distant metastasis (M0) (all P<0.05). The total number of CTC, mixed CTC and interstitial CTC were positively correlated with clinical stage, lymph node metastasis and distant metastasis (all P<0.05). RAS/RAF gene mutation, gender, age, tumor location and tumor differentiation did not affect the positive rate of CTC (all P>0.05). Conclusions:The results of CTC typing are of great research significance for comprehensive treatment, prognosis assessment and stratified management of CRC, among which the interstitial type of CTC may be a high risk factor for the recurrence and metastasis of CRC.

Clinical value of peripheral blood circulating tumor cells in the diagnosis and treatment of prostate cancer / 中国基层医药

Objective:To investigate the clinical value of peripheral blood circulating tumor cells (CTC) in the diagnosis and treatment of prostate cancer.Methods:Sixty-four patients with prostate cancer who received treatment in Xinjiang Production and Construction Corps Hospital, China between June 2018 and May 2020 were included in the cancer group. An additional 35 patients with benign prostatic lesions who concurrently received treatment in the same hospital were included in the benign disease group. Twenty male patients with non-prostate disease were included in the control group. Cell enrichment, separation, staining and identification together with Gleason score and pathological stage were subjected to one-way analysis of variance.Results:The percentage of patients with CTC count ≥ 3 in the cancer, benign disease and control groups was 73.43% (47/64), 17.14% (6/35) and 10.00% (2/20), respectively. The level of prostate-specific antigen in patients with CTC was significantly higher than that in patients without CTC ( t = 2.89, P < 0.05). There was significant difference in CTC count between different Gleason score groups ( F = 3.25, P < 0.05) and between different pathological stage groups ( F = 3.42, P < 0.05). Conclusion:Peripheral blood CTC measurement can be used as an auxiliary method for the differentiation of benign and malignant prostate diseases. CTC count in patients with prostate cancer is correlated with prostate-specific antigen level, Gleason score, and pathological stage. Therefore, peripheral blood CTC measurement plays an auxiliary role in predicting prognosis in patients with CTC. This study is innovative and scientific.

The combination of mammaglobin A and TWIST-1 increases detection of circulating tumor cells in breast cancer / A combinação de mamaglobina A e TWIST-1 aumenta a detecção de células tumorais circulantes no câncer de mama

ABSTRACT Objectives: Breast cancer cells that are released into the bloodstream are called circulating tumor cells (CTCs). CTCs can express different genes, like TWIST-1 and mammaglobin A (MGA). The aims of this study were to analyze the expression of TWIST-1 and MGA in the blood of breast cancer patients to detect CTCs and to assess the association between the presence of CTCs and prognostic parameters of breast cancer. Methods: Prospective study. Total ribonucleic acid (RNA) from blood mononucleated cells was obtained from breast cancer patients (n = 36; age: 51.5 ± 12.5 years) and healthy donors (n = 14; age: 49.4 ± 9.4 years). Real-time polymerase chain reaction (RT-PCR) was performed to analyze the expression of TWIST-1 and MGA. Results: Patient carcinomas - ductal (86.7%), other types (13.3%). MGA gene expression was not detected in the donors' samples, while it was detected in 14% of the patient samples. Overexpression of TWIST-1 gene was observed in 17% of the patient samples. The combined analysis of both markers allowed the detection of CTCs in 27.8% of the samples, resulting in a significant (p < 0.05) sensitivity increase of detection. No significant associations (p > 0.05) were found between expression of the analyzed genes and the breast cancer prognostic factors. Conclusion: Combined analysis of TWIST-1 and MGA increased the sensitivity of CTCs detection compared to the single analysis of each gene. The detection of CTCs was not associated with known prognostic factors, suggesting that it is able to provide clinical information in addition to routine breast cancer clinicopathological parameters.

RESUMEN Objetivos: Las células de cáncer de mama liberadas al torrente sanguíneo se llaman células tumorales circulantes (CTCs). Las CTCs pueden expresar diferentes genes, como TWIST-1 y mamaglobina A (MGA). Los objetivos de este estudio fueron analizar la expresión de TWIST-1 y MGA en la sangre de pacientes con cáncer de mama (CM) para detectar CTCs y evaluar la asociación entre la presencia de CTCs y los parámetros pronósticos del CM. Métodos: Estudio prospectivo. Se obtuvo el ácido ribonucleico (ARN) de las células mononucleadas en la sangre de pacientes con CM (n = 36, edad: 51,5 ± 12,5 años) y donantes sanas (n = 14; edad: 49,4 ± 9,4 años). Se realizó reacción en cadena de la polimerasa en tiempo real (RT-PCR) para analizar la expresión de TWIST-1 y MGA. Resultados: Carcinoma ductal (86,7%), otros tipos (13,3%). No se detectó la expresión del gen MGA en las muestras de las donantes, pero en el 14% de las muestras de las pacientes. Se observó elevada expresión de TWIST-1 en el 17% de las muestras de pacientes con CM. El análisis combinado de ambos marcadores permitió detección de CTCs en el 27,8% de las muestras, resultando en un aumento significativo (p < 0,05) en la sensibilidad de detección. No se encontraron asociaciones significativas (p > 0,05) entre la expresión de los genes y los factores pronósticos. Conclusión: El análisis combinado de TWIST-1 y MGA aumentó la sensibilidad de detección de CTCs en comparación con el análisis de cada gen. La detección de CTCs no se asoció a factores pronósticos conocidos, sugiriendo que podría ofrecer informaciones clínicas adicionales a los parámetros clínico-patológicos de rutina del CM.

RESUMO Objetivos: As células cancerígenas da mama liberadas na corrente sanguínea são chamadas de células tumorais circulantes (CTCs). As CTCs podem expressar diferentes genes, como TWIST-1 e mamaglobina A (MGA). Os objetivos deste estudo foram analisar a expressão de TWIST-1 e MGA no sangue de pacientes com câncer da mama (CM) para detectar CTCs e avaliar a associação entre a presença de CTCs e os parâmetros prognósticos do CM. Métodos: Estudo prospectivo. O ácido ribonucleico (RNA) das células mononucleadas no sangue foi obtido de pacientes com CM (n = 36, idade: 51,5 ± 12,5 anos) e doadoras saudáveis (n = 14; idade: 49,4 ± 9,4 anos). Reação da cadeia da polimerase em tempo real (RT-PCR) foi realizada para analisar a expressão de TWIST-1 e MGA. Resultados: Carcinoma ductal (86,7%), outros tipos (13,3%). A expressão do gene MGA não foi detectada nas amostras das doadoras, mas foi observada em 14% das amostras das pacientes. Superexpressão de TWIST-1 foi observada em 17% das amostras dos indivíduos com CM. A análise combinada de ambos os marcadores permitiu a detecção de CTCs em 27,8% das amostras, resultando em um aumento significativo (p < 0,05) na sensibilidade da detecção. Associações significativas (p > 0,05) entre a expressão dos genes e os fatores prognósticos não foram encontradas. Conclusão: A análise combinada de TWIST-1 e MGA aumentou a sensibilidade da detecção de CTCs em comparação com a análise de cada gene. A detecção de CTCs não foi associada a fatores prognósticos conhecidos, sugerindo que ela pode fornecer informações clínicas adicionais aos parâmetros clinicopatológicos de rotina do CM.

Proteínas mir-203a-3p e mmp-2 são altamente expressas em células tumorais circulantes de pacientes com carcinoma pancreático / Mir-203a-3p and mmp-2 proteins are highly expressed in circulating tumor cells from patients with pancreatic carcinoma

RESUMO -RACIONAL: O adenocarcinoma ductal do pâncreas é a quarta causa de morte associada ao câncer mais comum no mundo ocidental. A presença de células tumorais circulantes (CTCs) pode ser considerada um potencial fator prognóstico, visto que essas células representam a progressão tumoral, permitindo o monitoramento da eficácia terapêutica. OBJETIVOS: explorar as características morfológicas, moleculares e fenotípicas das células tumorais circulantes (CTCs) do sangue de pacientes com carcinoma pancreático e correlacionar os achados com a resposta ao tratamento, sobrevida livre de progressão, sobrevida global (SG) e trombose venosa profunda (TVP). MÉTODOS: o sangue periférico (10mL) foi analisado antes do início do tratamento e após 60 e 120 dias. As CTCs foram detectadas pelo ISET® e caracterizadas por imunocitoquímica. Para análise de miRNAs, leucócitos periféricos dos mesmos pacientes e indivíduos saudáveis foram coletados em paralelo no início do estudo. A expressão de miRNAs foi avaliada usando TaqMan T Array Human MicroRNA Cards v2.0. RESULTADOS: foram incluídos 9 pacientes. As proteínas MMP2 e TGFß-RI foram altamente expressas (77,7%) nas CTCs no início do estudo. No primeiro acompanhamento, MMP2 era predominante (80%) e no segundo acompanhamento, MMP2 e vimentina eram predominantes (50%). Microêmbolos tumorais circulantes (MTC) foram encontrados em dois pacientes e ambos apresentavam TVP. O miR-203a-3p foi altamente expresso em CTCs. miR-203a-3p está envolvido na estimulação da transição epitelio-mesenquima (TEM) e relacionado a pior SG no câncer pancreático (dados TCGA). CONCLUSÃO: Devido ao baixo número de pacientes e curto seguimento, não observamos correlação entre CTCs e resposta ao tratamento. No entanto, houve uma correlação entre MTC e TVP. Além disso, miR-203a-3p foi altamente expresso em CTCs, corroborando os achados de proteínas EMT. Este estudo abre perspectivas sobre a mudança dinâmica no padrão de proteínas expressas ao longo do tratamento e a utilização de miRNAs como novos alvos no carcinoma pancreático.

ABSTRACT - BACKGROUND: Ductal adenocarcinoma of the pancreas is the fourth most common cancer-associated cause of death in the Western world. The presence of circulating tumor cells (CTCs) can be considered a potential prognostic factor, as these cells represent tumor progression, allowing monitoring of therapeutic efficacy. OBJECTIVES: The objectives of this study were to explore the morphological, molecular, and phenotypic characteristics of CTCs from the blood of patients with pancreatic carcinoma and to correlate the findings with response to treatment, progression-free survival, overall survival (OS), and deep vein thrombosis (DVT). METHODS: Peripheral blood (10 mL) was analyzed before the beginning of treatment after 60 and 120 days. CTCs were detected by using ISET® and characterized by immunocytochemistry. For microRNAs (miRNAs) analysis, peripheral leukocytes from the same patients and healthy individuals (controls) were collected in parallel at baseline. The expression of miRNAs was evaluated (in pool) using TaqMan® Array Human MicroRNA Cards v2.0. RESULTS: Only nine patients were included. The proteins, namely, matrix metalloproteinase-2 (MMP2) and TGFβ-RI, were highly expressed (77.7%) in CTCs at baseline; at the first follow-up, MMP2 was predominant (80%) and, at the second follow-up, MMP2 and vimentin were predominant (50%). Circulating tumor microemboli (CTMs) were found in two patients and both presented DVT. The miR-203a-3p was highly expressed in CTCs. The miR-203a-3p is involved in the stimulation of epithelial-to-mesenchymal transition (EMT) and is related to worse OS in pancreatic cancer (TCGA data). CONCLUSION: Due to the low number of patients and short follow-up, we did not observe a correlation between CTCs and response to treatment. However, there was a correlation between CTM and DVT and also miR-203a-3p was highly expressed in CTCs, corroborating the findings of EMT proteins. This study opens the perspectives concerning the dynamic change in the pattern of proteins expressed along with treatment and the use of miRNAs as new targets in pancreatic carcinoma.

Identificação e correlação molecular e dinâmica de proteínas relacionadas à resistência ao tratamento neoadjuvante em células tumorais circulantes de pacientes com câncer de reto localmente avançado / Molecular and dynamic evaluation of proteins related to resistance to neoadjuvant treatment with chemoradiotherapy in circulating tumor cells of patients with locally advanced rectal cancer

INTRODUÇÃO: A quimiorradioterapia neoadjuvante (QRTN) consolidou-se como a principal estratégia para o tratamento do câncer de reto localmente avançado (CRLA). No entanto, respostas heterogêneas são observadas com o tratamento neoadjuvante, com apenas 15-20% dos pacientes com resposta patológica completa (RPC). Diante da necessidade de estratificar os pacientes em respondedores e não respondedores à QRTN antes do seu início, com o objetivo de aprimorar a seleção daqueles com maior probabilidade de obter uma RPC, vários estudos avaliam a identificação de possíveis biomarcadores. O objetivo primário deste estudo prospectivo foi analisar se a ausência da expressão do homólogo B de RAD23 (RAD23B) e da timidilato sintase (TYMS) nas células tumorais circulantes (CTCs) se correlacionaria com a RPC para os pacientes submetidos à QRTN e, assim, identificar possíveis respondedores ao tratamento. Os desfechos secundários foram avaliar a cinética das CTCs antes (C1) e após QRTN (C2), além da correlação da expressão de marcadores de resposta imune, como o Tumor Growth Factor ß Receptor I (TGF-ßRI) e Programmed Death ligand-1 (PD-L1) com a sobrevida livre de doença (SLD) e sobrevida global (SG). MÉTODOS: Entre 2016 e 2020, 63 pacientes com CRLA (cT3/T4 e/ou N+) submetidos a QRTN foram incluídos no estudo. As CTCs foram isoladas por ISET e avaliadas por imunocitoquímica. A expressão de RAD23B, TYMS, PD-L1 e TGF-ßRI foi avaliada nesta ordem de prioridade de acordo com o objetivo primário do estudo em cada momento de coleta e a disponibilidade de células (contagem de CTCs > 0) na amostra. RESULTADOS: Em C1, RAD23B foi detectado em 54,1% dos pacientes sem RPC e sua ausência em 91,7% dos pacientes com RPC (p = 0,014); Na segunda coleta, dos 13 pacientes com RPC, 10 não apresentaram expressão de RAD23B nas CTCs. Para os pacientes que não obtiveram RPC com QRTN, 51,7% apresentavam a expressão de RAD23B em CTC em C2 (p = 0,06). Na análise univariada (OR =0,077;IC 95%, 0,009-0,661; p = 0,019) e multivariada (OR= 0,064;CI 95%, 0,006-0,75; p = 0,029) para RPC, observamos que a expressão de RAD23B foi associado com menor chance de resposta em comparação com os pacientes com a ausência da expressão do RAD23B na C1. A ausência da expressão da TYMS foi observado em 90% dos pacientes com RPC e sua expressão em 51,7% sem RPC (p = 0,057). Na avaliação da cinética da CTCs pacientes com CTC2> CTC1 (cinética desfavorável) tiveram pior SLD (p = 0,00025) e SG (p = 0,0036) em comparação com aqueles com CTC2 ≤CTC1 (cinética favorável). A expressão de TGF-ßRI em qualquer momento das coletas correlacionou-se com pior SLD (p = 0,059). CONCLUSÃO: Demonstramos uma possível correlação entre a ausência de expressão de RAD23B e TYMS nas CTCs com a RPC, sendo um resultado importante para identificar os respondedores ao tratamento neoadjuvante, ajudando individualizar a abordagem terapêutica. Além disso, a cinética desfavorável e a expressão de TGF-ßRI nas CTCs se correlacionaram com pior sobrevida

INTRODUCTION: Neoadjuvant chemoradiotherapy (NCRT) has established itself as the main strategy for the treatment of locally advanced rectal cancer (LARC). However, heterogeneous responses are observed with neoadjuvant treatment, with only 15-20% of patients with complete pathological response (pCR). Given the need to stratify patients into responders and non-responders to NCRT prior to its initiation, in order to improve the selection of those most likely to obtain a pCR, several studies have assessed the identification of potential biomarkers capable of stratifying and monitoring the patient's response. The primary objective of this prospective study was to analyze whether the absence of RAD23 homolog B expression (RAD23B) and thymidylate synthase (TYMS) in circulating tumor cells (CTCs) would correlate with pCR for patients undergoing NCRT and thus identify possible responders to treatment. The secondary outcomes were to evaluate the kinetics of CTCs before (C1) and after NCRT (C2), in addition to the correlation of the expression of immune response markers, such as Tumor Growth Factor ß Receptor I (TGF-ßRI) and Programmed Death ligand- 1 (PD-L1) with clinical outcomes such as disease-free survival (DFS) and overall survival (OS). METHODS: Between 2016 and 2020, 63 patients (pts) with LARC (cT3 / T4 or N +) submitted to NCRT were included in the study. CTCs were isolated by ISET and evaluated by immunocytochemistry (protein expression). The expression of RAD23B, TYMS, PD-L1 and TGF-ßRI was evaluated in this order of priority according to the primary objective of the study at each time of collection (C1, C2 and C3) and the availability of cells (CTC count> 0) in the sample. RESULTS: In C1, RAD23B was detected in 54.1% of patients without pCR and its absence in 91.7% of patients with pCR (p = 0.014). In the second collection, of the 13 patients with pCR, 10 did not show RAD23B expression in the CTCs. For patients who did not obtain pCR with NCRT, 51.7% had RAD23B expression in CTC in C2 (p = 0.06). In the univariate (OR = 0.077; 95% CI, 0.009-0.661; p = 0.019) and multivariate (OR = 0.064; 95% CI, 0.006-0.75; p = 0.029) logistic regression models for pCR, we observed that the expression of RAD23B was associated with a lower chance of response compared to patients with the absence of RAD23B expression in C1. The absence of TYMS expression was observed in 90% of patients with pCR and its expression in 51.7% without pCR (p = 0.057). In the evaluation of CTCs kinetics patients with CTC2> CTC1 (unfavorable kinetics) had worse DFS (p = 0.00025) and overall survival (OS) (p = 0.0036) compared with those with CTC2 ≤CTC1 (favorable kinetics). TGF-ßRI expression at any time of the collections was correlated with worse DFS (p = 0.059). CONCLUSION: We demonstrated a possible correlation between the absence of RAD23B and TYMS expression in CTCs with pCR, being an important result to identify respondents to neoadjuvant treatment, helping to individualize the therapeutic approach. In addition, the unfavorable kinetics and expression of TGF-ßRI in CTCs correlated with worse survival.

Progress of circulating tumor cells in primary bone tumor / 肿瘤研究与临床

Some primary bone tumors are prone to hematogenous metastasis and after that, the therapeutic effect is not that good and prognosis is poor. Circulating tumor cells (CTC) shed from the tumor cells of primary or metastatic focus and then enter into blood circulation. CTC may appear in the early stage of the tumor, which can implant in distant organs to form metastatic sites and self-implant in the primary sites leading to the tumor recurrence; CTC are closely related with the prognosis of patients with tumors. In most primary bone tumors, CTC are heterogeneous compared with primary tumor cells. Studying CTC from various aspects can provide a basis for the early diagnosis and treatment of primary bone tumors. This review summarizes the current researches of CTC in common primary bone tumors, and expects the future of research direction and application practice in clinic.

Efficacy of folate-coupled quantum dots immunomagnetic beads method for detecting circulating tumor cells in epithelial ovarian cancer and the association of circulating tumor cells with clinicopathological features / 肿瘤研究与临床

Objective:To explore the effectiveness of folate-coupled quantum dots (FA-QD) immunomagnetic beads method for detecting circulating tumor cells (CTC) in epithelial ovarian cancer and the association of CTC with clinicopathological features of tumor patients.Methods:A total of 67 ovarian cancer patients in Shanxi Provincial People's Hospital from August 2019 to January 2020 were selected. Ovarian cancer SKOV-3 cells were divided into 5 cell number gradients (0, 100, 150, 200, 300), the detection rates of CTC were compared by using epithelial cell adhesion molecule (EpCAM) immunomagnetic beads (single standard method) and FA-QD immunomagnetic beads method (double standard method). The number of CTC in peripheral blood of ovarian cancer patients was detected by using FA-QD immunomagnetic beads method, and those with positive CTC under fluorescence microscope were treated as CTC positive patients. The association of CTC with clinicopathological factors and tumor markers of tumor patients was analyzed.Results:The average capture efficiency rate of CTC in SKOV-3 cells detected by FA-QD immunomagnetic beads method (83.4%) was higher than that by EpCAM immunomagnetic beads method (70.3%). Among 67 patients of ovarian cancer, the proportion of CTC positive patients was 30.0% (3/10) in stage Ⅰ-Ⅱ, 91.9% (34/37) in stage Ⅲ, 95.0% (19/20) in stage Ⅳ, and the difference was statistically significant ( P < 0.05). The proportion of CTC positive patients with lymph node metastasis was higher than that of patients without lymph node metastasis [97.1% (33/34) vs. 69.7% (23/33)], and the difference was statistically significant ( P < 0.05). The proportion of CTC positive patients with human epididymis protein 4 (HE4)>110 pmol/L was lower than that of patients with HE4 ≤ 110 pmol/L [58.8% (10/17) vs. 92.0% (46/50)], and the difference was statistically significant ( P = 0.005). There were no statistically significant differences in the proportion of CTC positive patients stratified by age, menopause, pathological differentiation, distant metastasis, carbohydrate antigen (CA) 125, CA199, carcino-embryonic antigen (CEA) (all P > 0.05). Conclusions:FA-QD immunomagnetic beads method can effectively detect CTC in peripheral blood of patients with epithelial ovarian cancer. The level of CTC in patients with epithelial ovarian cancer is related to lymph node metastasis, clinical TNM stage and HE4 level.

Correlation between the number of circulating tumor cells in peripheral blood and clinicopathological features of patients with breast cancer / 肿瘤研究与临床

Objective:To investigate the correlation between the number of circulating tumor cells (CTC) in peripheral blood and clinicopathological features of patients with breast cancer.Methods:The clinical data of 104 breast cancer patients at Guangzhou Panyu Central Hospital between January 2017 and May 2020 were retrospectively analyzed. The number of CTC in peripheral blood, the levels of serum tumor markers [alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen (CA)125, CA153] were detected. In blood samples, the number of CTC ≥ 2/ml was defined as CTC positive. Immunohistochemistry was used to analyze the protein expression of Ki-67 in tumor tissues. The association of CTC with clinicopathological features, serum tumor markers and Ki-67 protein expression was also analyzed.Results:The CTC positive rate was 80.77% (84/104). There were statistically significant differences in composition of whether there was vascular tumor thrombus (χ 2 = 0.860, P = 0.009), axillary lymph node metastasis (χ 2 = 12.382, P<0.01), N staging ( P = 0.002) and TNM staging (χ 2 = 7.698, P = 0.006) between patients with CTC positive and negative. However, there were no statistically significant differences in composition of age ( t = 0.634, P = 0.528), tumor quadrant (χ 2 = 6.523, P = 0.163), molecular subtyping (χ 2 = 4.164, P = 0.384), histological grade (χ 2 = 1.901, P = 0.387), T staging ( P = 0.099) and whether there was nerve invasion (χ 2 = 0.092, P = 0.761). The levels of serum CEA and CA125 in CTC positive patients were higher than those in CTC negative patients [median ( P25, P75): 2.50 ng/ml (2.21 ng/ml, 2.92 ng/ml) vs. 1.89 ng/ml (1.61 ng/ml, 2.35 ng/ml); 13.81 U/ml (11.79 U/ml, 16.28 U/ml) vs. 11.17 U/ml (8.91 U/ml, 12.80 U/ml); all P < 0.05], and CTC was positively correlated with serum CEA and CA153 levels ( r = 0.520, P<0.01; r = 0.497, P<0.01); CTC was not related to Ki-67 protein expression (χ 2 = 0.512, P = 0.474). Conclusion:The number of CTC in peripheral blood is closely related to clinical staging, lymph node or hematogenous metastasis, tumor markers CEA and CA153 levels of breast cancer. The increased number of CTC may cause tumor progression and metastasis.

Application progress of liquid biopsy in the diagnosis and treatment of biliary tract cancer / 中华肝胆外科杂志

With the development of precision medicine and individualized treatment, tissue biopsy in cancer patients diagnosis and therapy has been broadly used. However, because it’s hard to collect enough samples for biliary tract tumors, liquid biopsy was broadly applied for the diagnosis. In liquid biopsy, circulating tumor cells, circulating tumor DNA, and tumor-derived exosomes carrying tumor-specific information are released from tumor tissue into blood, bile, and other body fluids, which makes tumor biopsy samples easily to be obtained in a non-invasive way. At the same time, through a series of morphological and molecular measurements as well as genetic characterization, liquid biopsy can be used to look for the new early diagnostic markers, and therapeutic targets, monitoring progression and prognosis of diseases. This article outlined the current technology used to detect circulating tumor cells, circulating tumor DNA, and tumor-derived exosomes, and summarizes the latest advances in the clinical application of liquid biopsy in biliary tract cancers.

Values of serum neuron-specific enolase, circulating tumor cells and lactate dehydrogenase levels in the diagnosis and treatment of small cell lung cancer / 中国基层医药

Objective:To investigate the values of serum neuron specific enolase (NSE), circulating tumor cells (CTC) and lactate dehydrogenase (LDH) levels in the diagnosis and treatment of small cell lung cancer (SCLC).Methods:Ninety patients with SCLC who received treatment in the Second Affiliated Hospital of Zhejiang Chinese Medical University, China between December 2017 and December 2019 were retrospectively included in the observation group. Ninety healthy subjects who concurrently received lung examination in the same hospital were included in the healthy control group. An additional 90 patients with benign lung disease were included in the benign lung disease group. Serum NSE, CTC and LDH levels were determined in each group. The values of serum NSE, CTC and LDH levels in the diagnosis of SCLC were analyzed. Serum NSE, CTC and LDH levels were compared between before and after chemotherapy and their values in the treatment of SCLC were analyzed.Results:There were significant differences in serum NSE, CTC and LDH levels between three groups ( F = 359.789, 188.873 and 768.704, all P < 0.001). Serum NSE, CTC and LDH levels in the benign lung disease group were significantly greater than those in the healthy control group and significantly lower than those in the observation group. The receiver operating characteristic curve (ROC curve) analysis showed that the AUC values of serum NSE, CTC and LDH levels in the diagnosis of SCLC were 0.995, 0.953 and 0.987, respectively. The diagnostic accuracy was very high. The value at the maximum tangent point of Youden's index of serum NSE, CTC and LDH levels at the left-upper corner of the ROC curve was taken as the most appropriate cut-off value. The sensitivity and specificity of the most appropriate cut-off value of serum NSE, CTC and LDH levels in the prediction of SCLC were 100.0%/94.4%/91.1% and 94.4%/88.3%/100.0%, respectively. Therefore, serum NSE, CTC and LDH levels were of high values in the predication of SCLC. After chemotherapy, serum NSE, CTC and LDH levels in patients with SCLC were significantly lower than those before chemotherapy [NSE: (12.26 ± 3.26) μg/L vs. (18.36 ± 4.64) μg/L; CTC: (3.54 ± 1.08) counts/5 mL vs. (7.34 ± 1.30) counts/5 mL; LDH: (24.61 ± 9.66) U/L vs. (50.29 ± 16.29) U/L, t = 10.205, 12.864, 21.330, all P < 0.001). Serum NSE, CTC and LDH levels in SCLC patients in whom treatment was effective were significantly lower than those in SCLC patients in which treatment was not effective ( t = 8.111, 7.347, 10.731, all P < 0.001). Spearman correlation results showed that serum NSE, CTC and LDH levels were significantly negatively correlated with curative effects ( r = -0.562, -0.562, -0.758, all P < 0.05). Conclusion:Serum NSE, CTC and LDH levels are highly expressed in SCLC patients, which can be used as markers for early clinical diagnosis and treatment of SCLC.

Short-term efficacy of pemetrexed combined with cisplatin in the treatment of malignant pleural effusion and its effect on serum carbohydrate antigen 199 level and circulating tumor cells / 中国基层医药

Objective:To investigate the short-term efficacy of pemetrexed combined with cisplatin in the treatment of malignant pleural effusion and its effect on serum carbohydrate antigen 199 level and circulating tumor cells.Methods:Sixty patients with advanced lung cancer complicated by malignant pleural effusion who received treatment in Healthcare Group of Cixi Third People's Hospital, China from January 2017 to January 2020 were included in this study. They were randomly assigned to receive intrapleural injection of cisplatin (cisplatin alone group, n = 30) or intrapleural injection of cisplatin combined with intravenous injection of pemetrexed (cisplatin + pemetrexed group, n = 30) after thoracic drainage. Before and 1 month after treatment, pleural effusion was measured to evaluate clinical efficacy and improvement in quality of life. Serum carcinoembryonic antigen level, serum carbohydrate antigen 199 level and circulating tumor cells were determined. Adverse reactions during the treatment were recorded. Results:Total effective rate and the rate of improvement in quality of life in the cisplatin + pemetrexed group were 66.67% (20/30) and 70.00% (21/30), respectively, which were significantly higher than those in the cisplatin alone group [40.00% (12/30) and 43.33% (13/30), χ2 = 4.286, 4.344, both P < 0.05]. After treatment, serum carcinoembryonic antigen and serum carbohydrate antigen 199 levels in each group were significantly decreased compared with before treatment (both P < 0.05). Serum carcinoembryonic antigen and serum carbohydrate antigen 199 level in the cisplatin + pemetrexed group were (22.26 ± 5.13) ng/mL and (20.12 ± 4.35) U/mL, respectively, which were significantly lower than those in the cisplatin alone group [(31.64 ± 6.46) ng/mL, (28.07 ± 5.61) U/mL, t = 6.228, 3.134, both P < 0.05). In the cisplatin alone group, there was no significant difference in the proportion of circulating tumor cells before and after treatment ( P > 0.05). In the cisplatin + pemetrexed group, the proportion of circulating tumor cells after treatment was significantly lower than that before treatment ( χ2 = 4.286, P < 0.05). After treatment, there was no significant difference in the proportion of circulating tumor cells between the two groups ( P > 0.05). During the treatment, there were no significant differences in the incidences of rash, nausea and vomiting, leukopenia, and anemia between the two groups (all P > 0.05). Conclusion:Pemetrexed combined with cisplatin in the treatment of malignant pleural effusion exhibits better short-term efficacy than cisplatin alone. The combined therapy is more conducive to relieving clinical symptoms and improving the quality of life with higher safety than monotherapy.

Application of thymidine kinase 1 and circulating tumor cell in malignant solid tumor / 中国医师杂志

Thymidine kinase 1 (TK1) is associated with the occurrence of early malignant tumor,tumor metastasis,recurrence,and the changes of TK1 in the treatment can also reflect the sensitivity of tumor to treatment.Circulating tumor cells (CTC) are removed from the primary tumor or metastatic tumor to blood.The appearance of CTC in malignant tumors is usually related to the poor prognosis,such as metastasis and recurrence,short survival time and so on.The combination of TK1 and CTC can also be used as an independent predictor of survival.Now we analyse and summarize the role of TK1 and CTC in tumorigenesis and development.

Circulating tumor cell detection approaching to clinical application / 中华检验医学杂志

As "seeds" of tumor metastasis, circulating tumor cell (CTC) has important clinical application value in early diagnosis, immunotherapy and prognosis evaluation of tumors. With a deep understanding of CTC, its applied research has switched from cell enumeration to the molecular typing and single-cell sequencing. However, the standardization of CTC detection is still at a primary stage, opportunities and challenges coexist. This paper will review the current status and challenges in clinical applications of CTC detection, and make some suggestions for future development.

Diagnostic value of folate receptor-positive circulating tumor cell detection in subcentimeter pulmonary nodules / 肿瘤研究与临床

Objective@#To investigate the value of the folate receptor (FR)-positive circulating tumor cell (CTC) detection in the diagnosis of benign and malignant subcentimeter pulmonary nodules(the maximum diameter ≤10 mm).@*Methods@#Thirty-seven patients with subcentimeter pulmonary nodules (the chest CT showed the maximum diameter was ≤10 mm) in the Xuanwu Hospital of Capital Medical University from July to December 2018 were collected. Among them, 22 cases were diagnosed with early stage lung adenocarcinoma by postoperative pathological diagnosis and another 15 cases were benign lung lesion. Venous blood samples from these patients were collected before surgery and then utilized to detect FR+ CTC level (defined unit as FU/3 ml) by novel ligand-targeted polymerase chain reaction (LT-PCR), and the enzyme-linked immunosorbent assay was used to detect the levels of tumor markers, including carcinoembryonic antigen (CEA), neuron-specific enolase(NSE), cytokeratin 19 fragment CYFRA21-1, carbohydrate antigen 125 (CA125), CA199, pro-gastrin releasing peptide (pro-GRP), etc. The t-test was used to compare the measurement values between the groups. The CTC value 8.70 FU/3 ml described in the detection kit instruction was used as the threshold. The binary logistic regression was used to analyze the risk factors of malignant pulmonary nodules. The kappa consistency test was used to identify the consistency of the diagnosis results obtained by the FR+ CTC level and the pathological results of surgically resected specimens. The receiver operating characteristic curve (ROC) was drawn to evaluate the efficiency of each index for the diagnosis of benign and malignant subcentimeter pulmonary nodules.@*Results@#The level of FR+ CTC in patients with early stage lung cancer was higher than that in patients with benign lung lesion, and the difference was statistically significant [(11.0±3.0) FU/3 ml vs. (7.0±3.7) FU/3 ml, t=-3.327, P = 0.001]. The level of FR+ CTC was not related to the age, gender and smoking history of patients (all P>0.05). Logistic regression analysis indicated that high-level FR+ CTC was one of the risk factors for malignant pulmonary nodules (OR = 37.333, 95% CI 3.994-349.010, P = 0.002). The kappa consistency test indicated that the level of FR+ CTC used for the diagnosis of lung subcentimeter nodules presented a certain accuracy (κ = 0.627, P < 0.01). ROC illustrated that the FR+ CTC was better than CEA, NSE and CYFRA21-1 when it was used as an indicator for the diagnosis of malignant pulmonary nodules. The area under the curve(AUC) of FR+ CTC was 0.830 (95% CI 0.639-0.968), and the diagnostic sensitivity and specificity were 72.7% (95% CI 49.6%-88.4%) and 93.3% (95% CI 66.0%-99.7%), respectively. When FR+ CTC, CEA, NSE and CYFRA21-1 were combined for lung cancer diagnosis, the AUC, sensitivity and specificity were 0.776 (95% CI 0.614-0.938), 86.4% and 73.3%, respectively.@*Conclusion@#The detection of FR+ CTC has a high value in the diagnosis of benign and malignant subcentimeter pulmonary nodules.

Application of thymidine kinase 1 and circulating tumor cell in malignant solid tumor / 中国医师杂志

Thymidine kinase 1 (TK1) is associated with the occurrence of early malignant tumor, tumor metastasis, recurrence, and the changes of TK1 in the treatment can also reflect the sensitivity of tumor to treatment. Circulating tumor cells (CTC) are removed from the primary tumor or metastatic tumor to blood. The appearance of CTC in malignant tumors is usually related to the poor prognosis, such as metastasis and recurrence, short survival time and so on. The combination of TK1 and CTC can also be used as an independent predictor of survival. Now we analyse and summarize the role of TK1 and CTC in tumorigenesis and development .

Circulating Tumor Cell Number Is Associated with Primary Tumor Volume in Patients with Lung Adenocarcinoma / 결핵

Evaluating mmp-2 and tgfß-ri expression in circulating tumor cells of pancreatic cancer patients and their correlation with clinical evolution / Avaliação da expressão de mmp-2 e tgfß-ri em células tumorais circulantes de pacientes com câncer de pâncreas e sua correlação com evolução clínica

ABSTRACT Background: Metastasis is common in the diagnosis of pancreatic cancer, and the presence of epithelial-mesenchymal transition markers in circulating tumor cells may suggest worse prognosis. Aim: To correlate the number of circulating tumor cells (CTCs) in the peripheral blood of patients with a locally advanced or metastatic pancreatic tumor and the protein expression involved in epithelial-mesenchymal transition (EMT) in CTCs with clinical characteristics, progression-free survival (PFS) and overall survival (OS). Method: This was a prospective study conducted using peripheral blood samples collected at three different times. CTCs were quantified by the ISET test and analyzed by immunocytochemistry. Proteins involved in EMT (vimentin, TGFß-RI and MMP2) were analyzed in all CTCs. Results: Twenty-one patients were included. Median CTCs detected were 22, 20 and 8 CTCs/8 ml blood at baseline, first and second follow-up, respectively. No statistically significant correlation was found in correlating the number of CTCs and the evaluated clinical characteristics, PFS, or OS. There was no difference in PFS and OS among the EMT markers in the groups with and without markers. Conclusion: CTC analysis was not relevant in this sample for comparing clinical findings, PFS and OS in patients with pancreatic cancer. However, marker analysis in CTCs could be useful for the MMP-2 and/or TGFß-RI expression, as observed by the separate PFS curve.

RESUMO Racional: A metástase é comum no diagnóstico de câncer de pâncreas; presença de marcadores de transição epitélio-mesenquimal nas células tumorais circulantes (CTCs) podem sugerir pior prognóstico. Objetivo: Correlacionar o número de CTCs no sangue periférico de pacientes com tumor de pâncreas localmente avançado ou metastático e expressão de proteínas envolvidas na transição epitélio-mesenquimal (TEM) nas CTCs com características clínicas, sobrevida livre de progressão (SLP) e global (SG). Método: Estudo prospectivo realizado por meio de coletas de sangue periférico em três tempos distintos. As CTCs foram quantificadas pelo sistema ISET e analisadas por imunocitoquímica. Proteínas envolvidas na TEM (vimentina, TGFß-RI e MMP2) foram analisadas em todas as CTCs. Resultados: Foram incluídos 21 pacientes. A mediana de CTCs detectadas foi de 22, 20 e 8 CTCs/8 ml de sangue no baseline, primeiro e segundo seguimentos, respectivamente. Na correlação entre número de CTCs e as características clínicas levantadas, SLP, SG não houve correlação estatisticamente significante. Nos marcadores de TEM não houve diferença de SLP e SG entre os grupos que apresentaram e não apresentaram marcação. Conclusão: As CTCs não se mostraram relevantes na comparação dos achados clínicos, SLP e SG em pacientes com câncer de pâncreas. No entretanto, pode ser que para a análise de marcador seja útil, como observado pelas curvas separadas de expressão de MMP-2 e TGFß-RI nas CTCs.

Circulating tumor cells in pancreatic cancer patients: progression in the detection methods and clinical application / 中华外科杂志

Circulating tumor cells (CTC) disseminate from primary tumors by undergoing epithelial mesenchymal transition that allow their entry into the circulation to drive metastatic formation in pancreatic cancer patients.Technological advances in detection and characterization of CTC are conducive to the early diagnosis, differential diagnosis, monitoring disease progression and predicating the probability of canceration or the chemotherapeutic efficacy. Nowadays, detection methods of CTC can be based on immunomagnetic beads technique, cell filtration or microfluidic chips technology, but there are great differences in the sample throughput, CTC recovery rate, purity, and CTC viability among them.Owing to the dilemma in detection methods, the intrinsic relevance between the biological characteristics of CTC and clinical manifestations is still not exactly elucidated. By the improved methodology, next generation sequencing technology and exploring the technique for culturing CTC in vitro and establishing xenotransplanted tumor model in nude mice, more and more biological information will be revealed, and finally, individualized treatment is achieved.

Progress of circulating tumor cells in the diagnosis and treatment of esophageal cancer / 肿瘤研究与临床

Esophageal cancer is one of the malignant tumors with the highest morbidity and mortality in China. Its early clinical symptoms are not typical, and the patients are already in the middle and late stage when they go to see the doctor. The 5-year survival rate of these patients is low and the prognosis is poor. At present, the precision medicine is gradually being applied to the field of clinical oncology. The detection of circulating tumor cells (CTC) has the advantages of non-invasive, easy to obtain specimens, reflect the overall state of the tumor, real-time monitoring, etc. The CTC detection has gradually become a hot research direction. CTC is tumor cell detached from the primary or metastatic tumors and released into the peripheral blood circulation. CTC detection can dynamically reflect the status of tumors. Combined with the latest reports at home and abroad, this paper mainly reviews the application and limitations of CTC in the field of esophageal cancer.