Chareeporn AkekawatchaiSittiruk RoytrakulNarumon PhaonakropJanthima JaresitthikunchaiSarawut JitrapakdeeMahidol UniversityThammasat UniversityThailand National Center for Genetic Engineering and Biotechnology2020-06-022020-06-022020-01-01Methods in Molecular Biology. Vol.2138, (2020), 185-19319406029106437452-s2.0-85082562019https://repository.li.mahidol.ac.th/handle/123456789/56138© 2020, Springer Science+Business Media, LLC, part of Springer Nature. 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.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyChapterSCOPUS10.1007/978-1-0716-0471-7_11