Patent No. 5,955,289

Issued: September 21, 1999

Inventors: Xinfang Ma and Joseph A. Rininger, Ithaca, NY; Brian E. Johnson, Spencer, NY; and Debra S. Whiting, Valois, NY

Assignee: Paracelsian, Inc., Ithaca, NY

 In vivo and in vitro assays for the detection and quantification of substances, which are carcinogenic, but not genotoxic or mutagenic, by measuring a correlative change in cyclin dependent kinases (CDK).

A method and assay to determine whether a test compound or sample is a nongenotoxic carcinogen, wherein the compound or sample to be tested is added to a cyclin dependent kinase (CDK) assay system. The assay system of this invention can be a living organism, a cell culture or a cell lysate, as long as the assay system contains a cyclin dependent kinase (CDK). An increase in the tyrosylphosphorylation level of CDK indicates that the test compound is a nongenotoxic carcinogen, or that the test sample contains a nongenotoxic carcinogen.

This assay also detects nonmutagenic carcinogens and substances having a cell proliferation effect. The nongenotoxic carcinogens that can be identified through the assay include tumor promoters, chlorinated biphenyls, hormones, dioxins and peroxisome proliferators, among others. The assay system can be assembled in the form of a test kit for diagnostic and environmental testing.

The above assay could also be used to quantify the potency of a particular growth factor (peptide hormone). A peptide growth factor would be added to the assay system instead of a xenobiotic (foreign chemical) and otherwise the assay would proceed without modification.

The method and assay of the invention can also be used to determine the potential of a chemical as an antineoplastic agent by reversing the steps outlined above. Starting with a transformed cell or transformed cell lysate, a potential antineoplastic agent would be tested for the capacity of the chemical to put the cells into the G.sub.0 state. This capacity would be determined by quantifying the decrease in tyrosylphosphorylation of the CDK. The only other modification necessary to convert the assay for nongenotoxic carcinogens to one for antineoplastic agents is to grow the neoplastic cells in vitro in a full serum complement (20% serum containing medium)

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