Cell-free assay using oncogene-induced apoptosis to identify inhibitors of apoptosis in drug-resistant cells

A method of identifying a compound which inhibits apoptosis in transformed mammalian cells, comprising the steps of:

• a) combining
• 1) a compound to be assessed;
• and
• 2) a cellular extract obtained from 293 cells, wherein the cellular extract comprises apoptotic machinery and a pro-apoptotic signal capable of activating the apoptotic machinery, and wherein the apoptotic machinery and the signal are uncoupled;
• and
• b) maintaining the combination produced in a) under conditions in which the apoptotic machinery is coupled to the signal which allows apoptosis to occur, wherein the conditions do not include addition of an exogenous agent that activates the apoptotic machinery;
• and
• c) detecting inhibition of apoptosis in the presence of the compound to be assessed, wherein inhibition of apoptosis indicates that the compound to be assessed inhibits apoptosis in transformed cells.

A method of identifying a compound which inhibits apoptosis in transformed mammalian cells, comprising the steps of:

• a) combining
• 1) a compound to be assessed;
• and
• 2) a cellular extract obtained from fibroblast cells which express E1A, wherein the cellular extract comprises apoptotic machinery and a pro-apoptotic signal capable of activating the apoptotic machinery, and wherein the apoptotic machinery and the signal are uncoupled;
• and
• b) maintaining the combination produced in a) under conditions in which the apoptotic machinery is coupled to the signal which allows apoptosis to occur, wherein the conditions do not include addition of an exogenous agent that activates the apoptotic machinery;
• and
• c) detecting inhibition of apoptosis in the presence of the compound to be assessed, wherein inhibition of apoptosis indicates that the compound to be assessed inhibits apoptosis in transformed cells.