确定性侧向位移芯片在儿童横纹肌肉瘤循环肿瘤细胞检测方法的可靠性研究  

Reliability study of deterministic lateral displacement chip for the detection of circulating tumor cells in pediatric rhabdomyosarcoma

作　　者：郎会萍 杨楠麟 周哲蝶 刘宗彬 陈小文[1] 文飞球[1] 袁秀丽[1] LANG Huiping;YANG Nanlin;ZHOU Zhedie;LIU Zongbin;CHEN Xiaowen;WEN Feiqiu;YUAN Xiuli(Department of Hematology and Oncology,Shenzhen Children′s Hospital,Shenzhen 518000,China;Shenzhen Pediatric Clinical Branch,Shantou University Medical College,Shenzhen 518000,China;Shenzhen Ruige Biotechnology Co.Ltd,Shenzhen 518000,China)

机构地区：[1]深圳市儿童医院血液肿瘤科,深圳市518000 [2]汕头大学医学院深圳儿科临床分院 [3]深圳瑞格生物

出　　处：《中国小儿血液与肿瘤杂志》2024年第3期174-180,共7页Journal of China Pediatric Blood and Cancer

摘　　要：目的采用一种高效的循环肿瘤细胞(CTCs)捕获鉴定方式扩展CTCs的液体活检的临床应用。探究CTCs与横纹肌肉瘤(RMS)患儿临床特征的关系。方法将横纹肌肉瘤TE761细胞系混悬到外周血中,使用确定性侧向位移芯片(DLD-ΔChip),依据细胞尺寸快速无损分离富集肿瘤细胞,之后采用免疫荧光标记(DAPI+/CD45-/MyoD1+),在显微镜下鉴定TE761系细胞。检测该项技术对于肿瘤细胞的分离富集效率,以及检测RMS患儿外周血中CTCs数目并发掘与患者临床状况的相关性。结果DLD-ΔChip对于TE761系细胞的捕获率为94.0%,白细胞去除率为99.99%,红细胞去除率为100%。经DLD-ΔChip分离后的TE761系细胞活性为94.0%,与分离前的细胞增殖无统计学差异(t=2.21,P>0.05)。使用DLD-ΔChip测量RMS患儿外周血CTCs,RMS中危组和高危组外周血中的CTCs存在相关关系(U=289.00,P<0.05)。结论DLD-ΔChip分离富集CTCs效率高,分离后的CTCs活性高。RMS患儿中的CTCs在患者临床危险度分级、判断预后方面有良好的应用前景。Objective To expand the clinical application of liquid biopsy of circulating tumor cells(CTCs)with an efficient method for the capture and identification of CTCs.And to explore the relationship between CTCs and the clinical features of children with Rhabdomyosarcoma(RMS).Methods In this study,the rhabdomyosarcoma cell line TE761 was gradient suspended into peripheral blood using Deterministic Lateral Displacement Chip(DLD-ΔChip),and tumor cells were enriched by rapid non-destructive separation according to cell size using DLD-ΔChip,and then immunofluorescent labeling(DAPI+/CD45-/MyoD1+) was used to identify TE761 line cells under a microscope.This study verifies the isolation and enrichment efficiency of this technology for tumor cells,detects the number of CTCs in peripheral blood of children with RMS,explores the correlation of CTCs with the clinical condition of patients.Results DLD-ΔChip captures TE761 lines with 94.0%,leukocyte removal at 99.99%,and erythrocyte removal at 100%.The activity of TE761 line cells isolated by DLD-ΔChip was 94.0%,and there was no significant difference in proliferation compared with cells before isolation(t=2.21,P>0.05).Peripheral blood CTCs was measured in 17 children with RMS using DLD-ΔChip,and the risk grade of RMS was correlated with CTCs in peripheral blood(U=289.00,P<0.05).Conclusions DLD-ΔChip isolation and enrichment of CTCs has high efficiency and high activity of isolated CTCs.CTCs in children with RMS have good application prospects in clinical risk classification and prognosis.

关 键 词：循环肿瘤细胞 确定性侧向位移芯片 分离和检测 横纹肌肉瘤 

分 类 号：R738.7[医药卫生—肿瘤]