Profiling of human normal and cancer cell lines using phenotype microarray analysis (613.5)

Abstract

The aim of this research project was to study how women’s cancer cell lines respond to anti‐cancer agents and whether the mTOR pathway was being targeted in them. Five cancer cell lines were used‐ two ovarian (OV90, TOV112D), two uterine (MES‐SA, KLE), one breast (SKBR) and one non‐cancerous kidney (HEK293). BIOLOG Phenotype Microarray anti‐cancer panels were used to profile these cell lines. Cells were seeded on a variety of BIOLOG panels consisting of 96 different anti‐cancer agents for 24‐36h. Cell growth was examined using a MTT cell proliferation assay. Six reagents‐ berberine chloride, azathioprine, celastrol, gossypol, miltefosine, and etoposide, had the largest growth impact on the cell lines. Cell lines were then treated with 5µM of these reagents and then subjected to the InstantOne Elisa assay to determine activation status of a protein‐pS6 kinase in the mTOR pathway. Cancer cells from different tissues responded differently to the same anti‐cancer agent; HEK293 cells showed maximum resistance to treatment with these compounds. The reagents were then used at doses varying from 0.05‐5μM to conduct a dose response profile. Growth inhibition was observed at a concentration of 0.5μM and above, in a cell line specific manner. Appropriate controls were used in all experiments. Our collective findings indicate that growth inhibition observed in some cell lines is due to the inhibition of the mTOR pathway.Grant Funding Source: Supported by the California State University Program for Education and Research in Biotechnology