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

Otília Menyhárt, Hajnalka Harami-Papp, Saraswati Sukumar, Reinhold Schäfer, Luca Magnani, Oriol de Barrios, B. Györffy

Research output: Contribution to journalReview article

29 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)300-319
Number of pages20
JournalBiochimica et Biophysica Acta - Reviews on Cancer
Volume1866
Issue number2
DOIs
Publication statusPublished - Dec 1 2016

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Guidelines
Research Personnel
Neoplasms
RNA
Genomic Instability
Transcriptome
Fluorescence Microscopy
Immunoprecipitation
Energy Metabolism
Genes
Flow Cytometry
Cell Death
Immunohistochemistry
Cell Proliferation
Neoplasm Metastasis
Costs and Cost Analysis
Equipment and Supplies
Polymerase Chain Reaction

Keywords

  • Cancer
  • Cell culture
  • Flow cytometry
  • Fluorescence microscopy
  • Functional assays
  • Gene chips
  • Hallmark
  • Immunohistochemistry
  • In vitro methods
  • Multiplexing
  • Next generation sequencing
  • PCR

ASJC Scopus subject areas

  • Oncology
  • Genetics
  • Cancer Research

Cite this

Guidelines for the selection of functional assays to evaluate the hallmarks of cancer. / Menyhárt, Otília; Harami-Papp, Hajnalka; Sukumar, Saraswati; Schäfer, Reinhold; Magnani, Luca; de Barrios, Oriol; Györffy, B.

In: Biochimica et Biophysica Acta - Reviews on Cancer, Vol. 1866, No. 2, 01.12.2016, p. 300-319.

Research output: Contribution to journalReview article

Menyhárt, Otília ; Harami-Papp, Hajnalka ; Sukumar, Saraswati ; Schäfer, Reinhold ; Magnani, Luca ; de Barrios, Oriol ; Györffy, B. / Guidelines for the selection of functional assays to evaluate the hallmarks of cancer. In: Biochimica et Biophysica Acta - Reviews on Cancer. 2016 ; Vol. 1866, No. 2. pp. 300-319.
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