Detecting Carcinoma Cells in Peripheral Blood of Patients With Hepatocellular Carcinoma by Immunomagnetic Beads and RT-PCR

Guo, Junming; Yao, Feng; Lou, Yan-Lu; Xu, Chang-feng; Xiao, Bingxiu; Zhou, Wenhong; Chen, Jian; Hu, Yaoren; Liu, Zhong

doi:10.1097/01.mcg.0000247996.19710.f2

Cited by 27 publications

(20 citation statements)

References 28 publications

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“…Therefore, the CellSearch system is not appropriate for detection of CTCs in HCC patients. Given that there are currently no antibodies specific for HCC cell surface antigens, few reports have been published on the isolation of CTCs in HCC patients (10)(11)(12). a-Fetoprotein (AFP), a well-known marker for HCC, is expressed in the cytoplasm of HCC cells.…”

Section: Introductionmentioning

confidence: 99%

Isolation of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Using a Novel Cell Separation Strategy

Cao²,

Chen³

et al. 2011

Clinical Cancer Research

143

139

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Purpose: To establish a sensitive and specific isolation and enumeration system for circulating tumor cells (CTC) in patients with hepatocellular carcinoma (HCC).Experimental Design: HCC cells were bound by biotinylated asialofetuin, a ligand of asialoglycoprotein receptor, and subsequently magnetically labeled by antibiotin antibody-coated magnetic beads, followed by magnetic separation. Isolated HCC cells were identified by immunofluorescence staining using Hep Par 1 antibody. The system was used to detect CTCs in 5 mL blood. Blood samples spiked with Hep3B cells (ranging from 10 to 810 cells) were used to determine recovery and sensitivity. Prevalence of CTCs was examined in samples from HCC patients, healthy volunteers, and patients with benign liver diseases or non-HCC cancers. CTC samples were also analyzed by FISH.Results: The average recovery was 61% or more at each spiking level. No healthy, benign liver disease or non-HCC cancer subjects had CTCs detected. CTCs were identified in 69 of 85 (81%) HCC patients, with an average of 19 AE 24 CTCs per 5 mL. Both the positivity rate and the number of CTCs were significantly correlated with tumor size, portal vein tumor thrombus, differentiation status, and the disease extent as classified by the TNM (tumor-node-metastasis) classification and the Milan criteria. HER-2 gene amplification and TP53 gene deletion were detected in CTCs.Conclusion: Our system provides a new tool allowing for highly sensitive and specific detection and genetic analysis of CTCs in HCC patients. It is likely clinically useful in diagnosis and monitoring of HCC and may have a role in clinical decision making.

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Section: Introductionmentioning

confidence: 99%

Isolation of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Using a Novel Cell Separation Strategy

Cao²,

Chen³

et al. 2011

Clinical Cancer Research

143

139

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“…In addition to the methods discussed above which directly detect CTCs by cell surface antigens, researchers also determine specific mRNA expressed in cancer cells by RT-PCR in peripheral blood samples or after an initial enrichment [4,27]. If the results are positive, they indirectly suggest the presence of CTCs.…”

Section: Detection Of Tumor-specific Mrna Mirna and Tumor Cell-free Dnamentioning

confidence: 99%

“…in the CTC studies is that an extremely small number of CTCs exists in the bloodstream [4] and common serological, imaging and pathological approaches are not sensitive enough to effectively capture CTCs. Approximately less than 10 CTCs may be found among one billion blood cells; therefore highly sensitive methods are required to detect and isolate these cells from the bloodstream [1].…”

Section: How To Capture Ctcsmentioning

confidence: 99%

“…Whether tumor cells will be detected with this method depends on the number of cells to be analyzed. Given the fact that the number of CTCs in the peripheral blood of patients with liver cancer is often less than 10 per ml [4] or 2-8 per 10 9 blood cells [16], this device has a limited application in the capture of CTCs in a large volume of blood. However, it is a very useful research tool to isolate a highly pure pool of CTCs with specific surface markers, such as EpCAM or CD133 from samples that have been pre-enriched by microfiltration, gradient centrifugation or elutriation [16,17].…”

Section: Fluorescence-activated Cell Sortingmentioning

confidence: 99%

“…According to this principle, target cells in the patient's blood samples are separated by centrifugation with cell separation medium. The cell gradient medium often used for CTC separation is Ficoll [4][5][6]. Whole blood with anticoagulants is added directly into the centrifuge tubes.…”

Section: Isolation By Cell Densitymentioning

confidence: 99%

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Capturing circulating tumor cells of hepatocellular carcinoma

Wu¹,

Pan²,

Pei³

et al. 2012

Cancer Letters

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a b s t r a c tEarly metastases of hepatocellular carcinoma (HCC) may be detected by the isolation of circulating tumor cells (CTCs) in the bloodstream. During the course of therapeutic attempts, monitoring CTC changes in patients with HCC is helpful for the efficacy assessment. Nevertheless, the markers used for the detection, such as a-feto protein, asialoglycoprotein receptor or epithelial cell adhesion molecule, CD133 or CD90, are not specific for HCC CTCs. In spite of these limitations, a timely determination of the existence of CTCs will be beneficial for the monitoring of distant metastases, the evaluation of therapeutic attempts, and the prediction of prognosis.

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Aptamer‐Enabled Tumor Cell Isolation

Zhang

Fan

2016

Circulating Tumor Cells

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Detecting Carcinoma Cells in Peripheral Blood of Patients With Hepatocellular Carcinoma by Immunomagnetic Beads and RT-PCR

Abstract: Combining negative and positive immunomagnetic beads with RT-PCR technique may improve the sensitivity and specificity of detecting circulating HCC cells.

Cited by 27 publications

References 28 publications

Isolation of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Using a Novel Cell Separation Strategy

Isolation of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Using a Novel Cell Separation Strategy

Capturing circulating tumor cells of hepatocellular carcinoma

Aptamer‐Enabled Tumor Cell Isolation

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