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Simultaneous Measurement of RNA, DNA, and Protein Synthesis in Mouse Tumor and Reticuloendothelial Tissue Slices Using Glucose-6-³H as a Common Precursor¹

Surgical Professorial Unit, St. Bartholomew's Hospital, London, E. C. 1., England

The need for an in vitro cytotoxic drug sensitivity test covering a broad metabolic field has led to the development of a method for simultaneous assessment of RNA, DNA, and protein synthesis in vitro by mouse tumor and reticuloendothelial tissue slices using glucose-6-³H as a common precursor. ³H/¹⁴C ratios are given following incubation with both glucose-6-³H and glucose-6-¹⁴C, and the time course of ³H incorporation into protein and the nucleic acids by slices of mouse spleen, thymus, lymph nodes, and tumor is presented. The use of actinomycin D and puromycin shows that incorporation into protein, RNA, and DNA can be separately measured on the same sample. This is further supported by detailed examination of the hydrolysis products of protein and the nucleic acids. Results are discussed in relation to other methods of measuring nucleic acid and protein biosynthesis and to the known pathways of anabolic conversion of glucose to protein and nucleic acids.

¹ Supported by a generous grant from the Board of Governors of St. Bartholomew's Hospital.

Received 4/17/68. Accepted 10/ 7/68.

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