Levels of viral glycoprotein provide a dose-dependent measure of tumor cell inhibition

Abstract

A chemosensitivity assay utilizing small replicate Mm5mt/c1 C3H mammary tumor cell cultures was developed to determine whether changes in viral antigen expression and release into culture fluids could be utilized as an in vitro measure of chemotherapeutic drug effect. The 52, 000 MW viral envelope glycoprotein (gp52) of the mouse mammary tumor virus (MMTV) was measured in culture fluids of control and drug-treated cultures while cell density was simultaneously determined by cel staining and OD 664 mu determination. While extracellular gp52 levels and cell density both progressively increased over 72 hours for control cultures, treatments with doxorubicin resulted in dose-dependent declines in both parameters at 24, 48, and 72 hours. Comparison of doxorubicin dosages for 50% reduction (ED 50s) in both parameters (0.68uM and 1.1uM) revealed a similar coordinate reduction in both cell density and MMTVgp52. When gp52 levels were further examined as a general measure of effect for a broad spectrum of 6 drugs with differing mechanisms of action, coordinate declines in cell density and MMTVgp52 provided a time and dose-dependent dual measure of effect for each of the drugs tested. Coordinate declines resulted in the same following hierarchy of concentration-dependent drug potency: methotrezate >62; 5-fluorouracil >62; doxorubicin >62; N [ phosphonacetyl- L aspartic acid] (PALA) >62; cis-platinum >62; cyclophosphamide. The dual measures of therapeutic effect afforded by this assay argue for its use as an in vitro measure of effect for optimizing drug treatments.