Cytoplasm protein GFAP magnetic beads construction and application as cell separation target for brain tumors
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Journal of Nanobiotechnology Open Access
RESEARCH
Cytoplasm protein GFAP magnetic beads construction and application as cell separation target for brain tumors Yang Zhao1†, Feng Jiang1†, Qinhua Wang1†, Baocheng Wang1, Yipeng Han1, Jian Yang1, Jiajia Wang1, Kai Wang2, Junping Ao2, Xunxiang Guo3, Xiaofei Liang2,3* and Jie Ma1*
Abstract Background: It is very important to develop a highly efficient cerebrospinal fluid (CSF) detection system with diagnosis and prediction function, for which the detection of circulating tumor cells (CTCs) in CSF is a good choice. In contrast to the past use of epithelial EpCAM as CTCs separation target, a cytoplasm protein of GFAP antibody was first selected to construct highly-sensitive immunomagnetic liposome beads (IMLs). The validation and efficiency of this system in capturing CTCs for brain tumors were measured both in vitro and in vivo. The associations between the numbers of CTCs in patients with their clinical characteristics were further analyzed. Results: Our data show that CTCs can be successfully isolated from CSF and blood samples from 32 children with brain tumors. The numbers of CTCs in CSF were significantly higher than those in blood. The level of CTCs in CSF was related to the type and location of the tumor rather than its stage. The higher the CTCs number is, the more possibly the patient will suffer from poor prognosis. Genetic testing in GFAP CTC-DNA by sanger sequencing, q-PCR and NGS methods indicated that the isolated CTCs (GFAP+/EGFR+) are the related tumor cell. For example, the high expression of NPR3 gene in CSF CTCs was consistent with that of tumor tissue. Conclusions: The results indicated that GFAP-IML CTCs isolation system, combined with an EGFR immunofluorescence assay of antitumor marker, can serve as a brand-new method for the identification of CTCs for brain tumors. Via lumbar puncture, a minimally invasive procedure, this technique may play a significant role in the clinical diagnosis and drug evaluation of brain tumors. Keywords: Circulating tumor cell, Cytoplasm protein, Liquid biopsy, Tumor diagnosis, Brain tumor
*Correspondence: [email protected]; [email protected] † Yang Zhao, Feng Jiang and Qinhua Wang contributed equally to this work 1 Department of Pediatric Neurosurgery, Shanghai Xin Hua Hospital Affiliated To Shanghai Jiaotong University, School of Medicine, No. 1665 Kongjiang Road, Shanghai 200092, China 2 State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiaotong University School of Medicine, No. 25/Ln 2200 Xie Tu Road, Shanghai 200032, China Full list of author information is available at the end of the article
Background Liquid biopsy is considered as a promising technique to decipher the characteristics of malignant tumors [1, 2]. Apart from blood testing, it can be extended to monitor other body fluids, such as cerebrospinal fluid (CSF) for tumors in the central nervous system (CNS) [3] including astrocytoma, ependymoma, medulloblastoma and so on. Diverse clinical
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