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Isolation and Characterization of Methionine-Independent Clones from Methionine-Dependent Cancer Cells

  • Stacey L. Borrego
  • Da-Wei Lin
  • Peter KaiserEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1866)

Abstract

Unlike normal cells, transformed cells are unable to grow when methionine in the growth media is restricted. Reversion to methionine independence is a rare event in transformed and malignant cells. Methionine-independent revertants provide an excellent system to identify metabolic signatures and molecular characteristics associated with methionine dependency of transformed cells. Revertants maintain the genetic background and general growth behavior of the parental cell line, except that they proliferate under methionine restriction such as in methionine-free media supplemented with homocysteine. Here we describe a general approach to generate methionine-independent revertants using the example of the triple-negative breast cancer cell line MDA-MB-468. To validate and characterize reversion we describe assays to evaluate cell proliferation and anchorage-independent growth in soft agar.

Key words

Methionine Methionine restriction Methionine restriction resistance Methionine independence Homocysteine Anchorage independence 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biological ChemistryUniversity of California, IrvineIrvineUSA

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