Related Experiment Video
Updated: Dec 25, 2025
Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
Published on: September 19, 2018
Proteomic Analysis of the Anoikis-Resistant Human Breast Cancer Cell Lines
Chareeporn Akekawatchai1, Sittiruk Roytrakul2, Narumon Phaonakrop2
1Faculty of Allied Health Sciences, Department of Medical Technology, Thammasat University, Pathumtani, Thailand.
Abstract:
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.
Insights
Cancer cells acquire anoikis resistance to metastasize. This study details a method to create anoikis-resistant cell cultures and analyzes their protein profiles to understand underlying mechanisms of cancer cell survival and invasion.
Area of Science:
- Oncology
- Cell Biology
- Proteomics
Background:
- Anoikis resistance is crucial for cancer metastasis and invasion, leading causes of cancer mortality.
- The molecular mechanisms driving anoikis resistance in cancer cells remain poorly understood.
Purpose of the Study:
- To develop a protocol for generating anoikis-resistant cell cultures from nonmetastatic (MCF-7) and metastatic (MDA-MB-231) breast cancer cell lines.
- To investigate the molecular changes associated with anoikis resistance using proteomic analysis.
Main Methods:
- Cells were cultured on poly-2-hydroxyethyl methacrylate coated plates for 24 hours to induce anoikis resistance.
- Cell viability was assessed using trypan blue staining and annexin V assays.
- Proteomic analysis was performed on both cells and conditioned media to identify proteins involved in anoikis resistance.
Main Results:
- A protocol for preparing anoikis-resistant MCF-7 and MDA-MB-231 cell lines was successfully established.
- Proteomic analysis identified specific protein profiles associated with anoikis resistance in both cell lines and their surrounding media.
Conclusions:
- The developed protocol enables the study of anoikis resistance in cancer cells.
- Proteomic insights provide a foundation for understanding the molecular basis of cancer cell survival and potential therapeutic targets.
More Related Videos
08:08Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors
Published on: February 27, 2015
10:55Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract ECME-based Three-dimensional System
Published on: January 8, 2018