The NCI has developed a model of 60 cell lines to test the proliferation index of cancer cells in response to anticancer agents. Proliferation, though important, can only tell part of the story, as the mechanism for this apparent lack of growth is missing. High content analysis is the ideal platform to investigate mechanism along with proliferation studies as data can be multiplexed within the same well. We have taken 6 cell lines in six different tumor origins to investigate the effects of sixteen different therapeutics on proliferation, apoptosis, necrosis, and cell cycle inhibition. The cell lines were chosen based on their ability to undergo two to three doublings in 72 hours and all were grown in standardized media and serum. These agents were selected for their role in signaling and anti-proliferative effects and compared between and within cellular origin along with some available genetic analysis. These mechanism of anti-proliferation can be investigated by use of nuclear count (proliferation), anti-activated caspase 3 antibodies (apoptosis), anti-phosphohistone H3 (cell cycle), and membrane leaking (necrosis/late apoptosis) within the same well, increasing consistency by enabling the normalization of data within a well. In addition, other cellular parameters such as changes in nuclear size can also be visualized and quantitated to add to the total understanding of the mechanisms of agents and their combinations. In conclusion, High Content Analysis coupled with genetic analysis is a robust and sensitive manner to be able to detail mechanism and sensitivity to chemotherapeutics agents.

98th AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA