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Potdar et al. J Cancer Metastasis Treat 2017;3:6-15 Journal of
DOI: 10.20517/2394-4722.2016.53
Cancer Metastasis and Treatment

www.jcmtjournal.com
Topic: Circulating Tumor Cells: Diagnostics and Clinical Applications Open Access

Profiling of circulating tumor cells in liquid

biopsies from metastatic cancer patients

Pravin D. Potdar, Keerti Sen

Department of Molecular Medicine and Biology, Jaslok Hospital and Research Centre, Mumbai 400026, Maharashtra, India.
Correspondence to: Dr. Pravin D. Potdar, Department of Molecular Medicine and Biology, Jaslok Hospital and Research Centre, 15 Dr. G.
Deshmukh Marg, Mumbai 400026, Maharashtra, India. E-mail: ppotdar@jaslokhospital.net; ppravin012@gmail.com
How to cite this article: Potdar PD, Sen K. Profiling of circulating tumor cells in liquid biopsies from metastatic cancer patients. J Cancer Metastasis
Treat 2017;3:6-15.
Dr. Pravin D. Potdar’s present interest is to study molecular profiling of circulating tumor cells (CTC), circulating
tumor DNA, cancer associated fibroblasts and cancer stem cells involved in metastatic process of cancers,
and to see how this process can be reverted back to normal by using innovated technologies which include
nanotechnology and nanomedicine.

ABSTRACT
Article history: Aim: Circulating tumor cells (CTCs) are crucial to tumor metastasis and valuable for
Received: 02-09-2016 prediction of clinical outcome in patients with solid tumors. Here, the authors aimed to
Accepted: 13-01-2017 establish a method for enumeration and characterization of CTCs from liquid biopsies.
Published: 23-01-2017 Methods: Peripheral blood mononuclear cells (PBMCs) were separated from blood samples
from patients with metastatic cancer using Ficoll-Hypaque gradients and cultured to isolate
Key words: and enumerate CTCs. Cultured CTCs were morphologically characterized by light and
Circulating tumor cells, phase contrast microscopy. The tumorigenicity of Ficoll-Hypaque-separated PBMCs was
liquids biopsies, examined, in addition to their expression of mRNA metastasis markers. Results: CTCs
molecular markers, were isolated in culture and enumerated by counting under phase contrast microscopy,
soft agar assay, demonstrating that 0.01-0.04% of total PBMCs were CTCs. CTCs were dormant, with
metastasis, large, oval-shaped, spiky morphology. PBMCs obtained from liquid biopsies exhibited
metastatic genes, anchorage-independent growth, forming numerous colonies in soft agar assays. Molecular
epithelial mesenchymal transition, profiling demonstrated expression of several metastatic genes, but not of cadherin 1
tumorigenic (encoding the adhesion protein), in all patients. Conclusion: The authors successfully
isolated, enumerated, and characterized CTCs from liquid biopsies of metastatic cancer
patients. This study has potential to facilitate the development of new diagnostic and
therapeutic methods using liquid biopsies, for application in metastatic cancers.

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