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Acquisition of anoikis resistance is a prerequisite for cancer metastasis and invasion, which are major causes of death from cancer. The molecular mechanisms underlying antianoikis properties in cancer cells are still largely unclear. Here, we describe a protocol for preparation of anoikis-resistant cultured nonmetastatic MCF-7 and metastatic MDA-MB-231 cell lines. The anoikis-resistant cultures were prepared by plating cells in the poly-2-hydroxyethyl methacrylate coated plates and cultured for 24 h. The viability of cells in the cultures was determined using trypan blue staining and annexin V cell death assay, while protein profiles associated with anoikis-resistance in both cells and conditioned media were analyzed by proteomics.

Citation

Chareeporn Akekawatchai, Sittiruk Roytrakul, Narumon Phaonakrop, Janthima Jaresitthikunchai, Sarawut Jitrapakdee. Proteomic Analysis of the Anoikis-Resistant Human Breast Cancer Cell Lines. Methods in molecular biology (Clifton, N.J.). 2020;2138:185-193

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PMID: 32219748

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