Guidelines for the selection of functional assays to evaluate the hallmarks of cancer

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Title: Guidelines for the selection of functional assays to evaluate the hallmarks of cancer
Authors: Menyhárt, O
Harami-Papp, H
Sukumar, S
Schäfer, R
Magnani, L
De Barrios, O
Győrffy, B
Item Type: Journal Article
Abstract: The hallmarks of cancer capture the most essential phenotypic characteristics of malignant transformation and progression. Although numerous factors involved in this multi-step process are still unknown to date, an ever-increasing number of mutated/altered candidate genes are being identified within large-scale cancer genomic projects. Therefore, investigators need to be aware of available and appropriate techniques capable of determining characteristic features of each hallmark. We review the methods tailored to experimental cancer researchers to evaluate cell proliferation, programmed cell death, replicative immortality, induction of angiogenesis, invasion and metastasis, genome instability, and reprogramming of energy metabolism. Selecting the ideal method is based on the investigator's goals, available equipment and also on financial constraints. Multiplexing strategies enable a more in-depth data collection from a single experiment — obtaining several results from a single procedure reduces variability and saves time and relative cost, leading to more robust conclusions compared to a single end point measurement. Each hallmark possesses characteristics that can be analyzed by immunoblot, RT-PCR, immunocytochemistry, immunoprecipitation, RNA microarray or RNA-seq. In general, flow cytometry, fluorescence microscopy, and multiwell readers are extremely versatile tools and, with proper sample preparation, allow the detection of a vast number of hallmark features. Finally, we also provide a list of hallmark-specific genes to be measured in transcriptome-level studies. Although our list is not exhaustive, we provide a snapshot of the most widely used methods, with an emphasis on methods enabling the simultaneous evaluation of multiple hallmark features.
Issue Date: 11-Oct-2016
Date of Acceptance: 8-Oct-2016
ISSN: 0304-419X
Publisher: Elsevier
Start Page: 300
End Page: 319
Journal / Book Title: Biochimica et Biophysica Acta-Reviews on Cancer
Volume: 1866
Issue: 2
Copyright Statement: © 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (
Keywords: Cancer
Cell culture
Flow cytometry
Fluorescence microscopy
Functional assays
Gene chips
In vitro methods
Next generation sequencing
Oncology & Carcinogenesis
06 Biological Sciences
02 Physical Sciences
1112 Oncology And Carcinogenesis
Publication Status: Published
Conference Place: Netherlands
Appears in Collections:Division of Surgery
Division of Cancer

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