Atomic Force Microscopy in Cancer Cell Research: Study of Phenotypic Changes Induced by Glycerol-3-phosphate acyltransferase-2 expression.

BIOQ. PRIETO E.D; CATTANEO, ELIZABETH; FASBIANI, M. BELEN; GONZALEZ BARÓ, M. DEL ROSARIO

La Plata, Buenos Aires

Workshop; Imaging Techniques for Biotechnology and Biomedical Applications Workshop; 2016

INIFTA-CINDEFI

Introduction: in mammalian cells, de novo synthesis of glycerolipids begins with the acylation of glycerol-3-phosphate, catalyzed by glycerol-3-phosphate acyltransferase (GPAT). (1) GPAT2 is a mitochondrial isoform primarily expressed in testis under physiological conditions, and overexpressed in several types of cancer and cancer-derived human cell lines, where its expression contributes to the tumor phenotype. (2) Using a gene silencing model and atomic force microscopy (AFM) technology we studied the correlation between GPAT2 expression and cell surface topography, roughness and membrane permeability in MDA-MB-231 cell line. In addition, we analized the glycerolipid composition by gas-liquid chromatography. Results: we observe that GPAT2 expressing cells exhibit a more roughness topography and less membrane damage that GPAT2 silenced cells. In this sense, pore-like structures were only observed in GPAT2 silenced cells, correlating with a higher membrane damage evidenced by lactate dehydrogenase release. These results suggest a role of GPAT2 in the regulation of these cell surface properties.Conclusions: cancers are one of the leading causes of morbidity and mortality worldwide, with approximately 14 million new cases and 8.2 million cancer related deaths in 2012 (http://www.who.int/mediacentre/factsheets/fs297/en/). In the last few years, AFM has been used in biomedical cancer research for early diagnosis of cancer, and for the study of cancer progression and metastasis (3, 4, 5) (Cross SE et al. 2007; Lekka M 2012; Kaul-Ghanekar R et al. 2009)GPAT2 protein expression induces changes in the cell surface that correlates with its contribution to the tumor phenotype, and might be used as a phenotypic differentiation marker. AFM provides the basis for the identification and quantification of these changes, and demonstrate the utility of this technique in the study of cancer cell biology.