The Relationship Between Circulating Tumor Cell And The Diagnosis、 Treatment Response Monitoring And Prognosis In Neuroblastoma

Objective: By dynamically monitoring changes in circulating tumor cell(CTC),to investigate the relationship between CTC and clinicopathological features of neuroblastoma(NB),and to evaluate the value of CTC in the early diagnosis,treatment response monitoring,and prognosis evaluation of NB.Methods: Seventy-three patients diagnosed with NB in the Shenzhen Children’s Hospital were enrolled in the study between April 2019 and August 2022.During the same period,15 healthy children were selected as the control group.We collected 2m L of peripheral venous blood from each patient at the initial diagnosis and after the first to sixth cycles of chemotherapy respectively.CTC was isolated and enriched by Cascaded filter deterministic lateral displacement microchips and was identified by immunofluorescence staining.Basic information,clinical and pathological data,and follow-up data were collected.The chi-square test,non-parametric test,receiver operating characteristic(ROC)curve,survival analysis,and other statistical methods were used for statistical analysis.The relationship between CTC at diagnosis and the clinicopathological features of NB was analyzed.The value of CTC in the diagnosis of metastasis was assessed.The correlation between clinical biological markers and CTC was observed.The relationship between CTC and the efficacy and prognosis of NB was analyzed.Results:1 CTC counts were detected in 54 NB children at diagnosis,among which 43 were positive.The positive detection rate was 79.6% and the median CTC was 5 per 2m L.One of the 15 healthy controls was positive,the count was 1per 2m L.The positive rate of CTC and the count of CTC in NB were significantly higher than those in healthy controls(P<0.001;P<0.001).2 There was no significant difference in CTC count between genders,ages,and MYCN gene status(P=0.721;P=0.499;P=0.176).There were statistically significant differences in CTC count among different primary sites,different INPC tissue types,and different INSS and INRG stages(P=0.005;P=0.001;P=0.001;P<0.001).The CTC count at diagnosis was 1 per 2m L in low-risk NB patients,3 per 2m L in intermediate-risk NB patients,and 12 per 2m L in high-risk patients.The higher the risk classification,the more the CTC count in peripheral blood(P=0.001).Patients with bone marrow metastasis,bone metastasis,and other site metastasis had significantly higher CTC count at diagnosis than those without metastasis(P<0.001;P<0.001;P=0.002).3 The positive rate of CTC was significantly different among different primary sites,INPC tissue types,and different INSS and INRG stages(P=0.009;P=0.006;P=0.006;P=0.002).The higher the risk classification,the higher the CTC positive rate at diagnosis(P=0.013).The CTC positive rate was significantly higher in NB patients with bone marrow metastasis and other site metastasis than those without metastasis(P=0.001;P=0.022).4 After two cycles of chemotherapy,the CTC count decreased significantly compared with the CTC count at diagnosis(P=0.013).5 The median number of CTC in NB patients with metastasis at diagnosis was14 per 2m L,which was significantly higher than 1.5 per 2m L in nonmetastatic patients(P < 0.001).NSE and CTC were included in the logistic regression equation,and the OR value of CTC was 14.684.CTC was an independent risk factor for metastasis.The area under the ROC curve(AUC)was 0.824.The optimal cut-off value was 6 per 2m L,with a sensitivity of68.8% and a specificity of 94.4%.The AUC of the combined diagnosis of NSE and CTC was 0.958.The sensitivity and specificity were 91.2% and94.4%,respectively.6 CTC count was positively correlated with bone marrow flow cytometry,NSE,and LDH(P<0.001).7 CTC count increased before clinical diagnosis of NB progression or recurrence,earlier than NSE and imaging changes.8 There was no statistical difference in PFS and OS between CTC-positive NB patients and CTC-negative NB patients at diagnosis(P=0.589;P=0.692).conclusion:1 Cascaded filter deterministic lateral displacement microchips have high sensitivity and specificity for detecting CTC in the peripheral blood of NB patients at diagnosis.2 The positive rate of CTC and the count of CTC at diagnosis were related to the clinical and pathological features of NB.3 The count of CTC decreased significantly after the second course of chemotherapy.CTC count is an independent risk factor for NB metastasis.Metastasis is more likely when CTC counts over 6 per 2m L at diagnosis.Combining CTC with NSE can improve diagnostic performance.4 CTC is helpful for early diagnosis of NB and monitoring the progression in NB.