EGFR Transgene Stimulates Spontaneous Formation of MCF7 Breast Cancer Cells Spheroids with Partly Loss of HER3 Receptor

• Published in: International journal of molecular sciences 2021-11, Vol.22 (23), p.12937
• Main Authors: Troitskaya, Olga, Novak, Diana, Nushtaeva, Anna, Savinkova, Maria, Varlamov, Mikhail, Ermakov, Mikhail, Richter, Vladimir, Koval, Olga
• Format: Article
• Language: English
• Subjects: 3D cell culture; Adenocarcinoma; Adhesives; Breast cancer; cancer stem cells; CD24 Antigen - metabolism; CD44 antigen; Cell culture; Cell Culture Techniques, Three Dimensional; Conversion; Drug development; Drug resistance; EGFR; Epidermal growth factor; Epidermal growth factor receptors; ErbB-2 protein; Genes, erbB-1; Growth conditions; Growth factor receptors; Growth factors; HER3; Humans; Hyaluronan Receptors - metabolism; Invasiveness; Kinases; MCF-7 Cells; MCF7; Metastasis; Phenotypes; Population; Receptor, ErbB-3 - metabolism; Rhodamine 123; Spheroids; Spheroids, Cellular; Stem cells; Suspension culture; Transgenes; Tumor Cells, Cultured; Tumors
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms222312937

Multicellular spheroids with 3D cell-cell interactions are a useful model to simulate the growth conditions of cancer. There is evidence that in tumor spheroids, the expression of various essential molecules is changed compared to the adherent form of cell cultures. These changes include growth factor receptors and ABC transporters and result in the enhanced invasiveness of the cells and drug resistance. It is known that breast adenocarcinoma MCF7 cells can spontaneously form 3D spheroids and such spheroids are characterized by high expression of EGFR/HER2, while the natural phenotype of MCF7 cells is EGFR /HER2 . Therefore, it was interesting to reveal if high epidermal growth factor receptor (EGFR) expression is sufficient for the conversion of adherent MCF7 to spheroids. In this study, an MCF7 cell line with high expression of EGFR was engineered using the retroviral transduction method. These MCF7-EGFR cells assembled in spheroids very quickly and grew predominantly as a 3D suspension culture with no special plates, scaffolds, growth supplements, or exogenous matrixes. These spheroids were characterized by a rounded shape with a well-defined external border and 100 µM median diameter. The sphere-forming ability of MCF7-EGFR cells was up to 5 times stronger than in MCF7 cells. Thus, high EGFR expression was the initiation factor of conversion of adherent MCF7 cells to spheroids. MCF7-EGFR spheroids were enriched by the cells with a cancer stem cell (CSC) phenotype CD24 /CD44 in comparison with parental MCF7 cells and MCF7-EGFR adhesive cells. We suppose that these properties of MCF7-EGFR spheroids originate from the typical features of parental MCF7 cells. We showed the decreasing of HER3 receptors in MCF7-EGFR spheroids compared to that in MCF and in adherent MCF7-EGFR cells, and the same decrease was observed in the MCF7 spheroids growing under the growth factors stimulation. To summarize, the expression of EGFR transgene in MCF7 cells stimulates rapid spheroids formation; these spheroids are enriched by CSC-like CD24 /CD44 cells, they partly lose HER3 receptors, and are characterized by a lower potency in drug resistance pomp activation compared to MCF7 . These MCF7-EGFR spheroids are a useful cancer model for the development of anticancer drugs, including EGFR-targeted therapeutics.