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The isolation and characterisation of circulating tumour cells from patients with hepatocellular carcinoma

Lookup NU author(s): Dr Sheba Macheka, Dr David Jamieson, Professor Helen Reeves, Misti McCain

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This is the final published version of a conference proceedings (inc. abstract) that has been published in its final definitive form by International Institute of Anticancer Research, 2016.

For re-use rights please refer to the publisher's terms and conditions.


Abstract

Background: Hepatocellular carcinoma (HCC) is the secondmost common cause of cancer-related mortality. Thecharacterization of circulating tumour cells (CTCs) fromperipheral blood, may provide information that may help predictprognosis or stratify patients for targeted therapies.Asialoglycoprotein receptor-1 (ASGPR1) is predominantlyexpressed by hepatocytes and this may be utilised as abiomarker to detect liver-specific CTCs. Some CTCs undergothe epithelial-to-mesenchymal-transition (EMT) and expressVimentin. This additional biomarker could aid with identifyingCTCs. Aims: To detect CTCs in blood samples from patientswith HCC, by targeting cells expressing ASGPR1 or Vimentinusing imagestreamX flow cytometry. Materials and Methods:Blood samples were obtained from 8 consenting patients withHCC. Red blood cells and leucocytes were depleted, in order toenrich for CTCs. Following this, patient samples were stainedwith a panel of antibodies conjugated with fluorochromes, inorder to identify Vimentin, Cytokeratin or ASGPR1 positiveCTCs using ImagestreamX. Furthermore, CTC detection wasbased on additional parameters such as surface area (>200μm2), the absence of CD45+ expression and a large nuclearcontent. Results: ASGPR1 and Cytokeratin were the mostcommon biomarkers expressed by CTCs detected in 5 out of 8of patients. A greater number of CTCs detected, expressed onlyvimentin compared to cytokeratin. This confirmed that CTCsdown-regulated the expression of epithelial biomarkers such ascytokeratin, due to EMT. Discussion: We designed a novelsensitive panel of antibodies that detected biomarker positiveCTCs which expressed ASGPR1, vimentin and cytokeratinfrom peripheral blood samples, using imagestreamX.


Publication metadata

Author(s): Macheka S, Jamieson D, Reeves H, McCain M

Publication type: Conference Proceedings (inc. Abstract)

Publication status: Published

Conference Name: 6th Christie International Student Cancer Conference

Year of Conference: 2016

Pages: 6717-2718

Online publication date: 01/12/2016

Acceptance date: 20/10/2016

Date deposited: 25/08/2017

ISSN: 1791-7530

Publisher: International Institute of Anticancer Research

URL: http://www.christie.nhs.uk/media/4536/the-christie-international-student-cancer-conference.pdf

Series Title: Anticancer Research


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