Journal: PLoS ONE

Article Title: Quantification of Dynamic Morphological Drug Responses in 3D Organotypic Cell Cultures by Automated Image Analysis

doi: 10.1371/journal.pone.0096426

Figure Lengend Snippet: The heatmaps illustrate changes in spheroid growth (A: Area) and general symmetry (B: Roundness) in response to the 19 compound treatments (5031–16415 multicellular structures analysed for each cell line). (A) CCG-1423, KH7, latrunculin A and narciclasine are preferentially cytotoxic and/or antiproliferative compounds across all cell lines, as highlighted by red boxes. Paclitaxel, at a concentration of 5 nM, shows partial cytotoxic/antiproliferative effects only in some of the cell lines. (B) Effects of mainly anti-invasive compounds IPA3 or NSC23766 were reproducible in many of the spontaneously invasive (or branching) cell lines PC-3, PC-3M Pro4, and RWPE-1. (C) Images segmented and analysed with AMIDA. Effective anti-invasive functions of the compounds IPA3, BPIPP and NSC23766 against the most aggressive, motile and invasive cell lines PC-3M, ALVA31, MDA-MB-231 (both SA and parental ATCC), and RWPE1. The extremely invasive PC3-derivative ALVA31 was not affected, however. (D) Blebbistatin and Y-27632 show invasion-inducing function in spheroids formed by the LNCaP and DU145 lines which typically form round spheroids and lack invasive properties.

Article Snippet: All compounds were purchased from Tocris (Bristol, UK), except for Ki-16425 (Cayman Chemical Company, Ann Arbor, MI, USA) and CCG-1423, Rac inhibitor I (#553502) and Rac inhibitor II (#553511) (Merck Millipore, Darmstadt, Germany), and dissolved in appropriate vehicle (DMSO, EtOH or PBS).

Techniques: Concentration Assay