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Studies on the Proteins of Rhabdomyosarcoma and Normal Muscle of Mice

I. Gross Composition, Extractions with 0.5 M KCl, and Fractionations by Differential Centrifugation and Dialysis

(From the Institute for Cancer Research and the Lankenau Hospital Research Institute, Philadelphia 11, Pennsylvania)

The production of a rhabdomyosarcoma in mice and its preparation for chemical analysis have been described.

Preliminary studies of gross composition of rhabdomyosarcoma and normal muscle revealed that the moisture content of rhabdomyosarcoma was significantly higher than that of normal muscle, while the reverse held true for the nitrogen contents. The phosphorus and ash contents of tumor tissue were higher than those of normal tissues when calculated on the basis of dry tissue, but no differences were observed on a wet basis. The significance of these findings was discussed.

Extraction studies made with a salt solution containing 0.45 M KCl, 0.05 M KHCO₃, and 0.001 M Na₄P₂O₇ showed that the nitrogen of the tumor was more readily and completely extracted than that of the normal tissue. Other differences in behavior of the tissues during extraction were noted.

Fractionation of extracts by differential centrifugation yielded appreciable amounts of "heavy" material from tumor but very little from normal muscle. Tests for ribose nucleic acid and desoxyribose nucleic acid contents indicated that the heavy material obtained from a first extraction of tumor tissue represented largely cytoplasmic nucleoprotein, while heavy material from subsequent extracts represented primarily nuclear nucleoprotein.

Reduction of salt concentration of the extracts by means of dialysis, which yielded myosin from normal muscle, yielded also a myosin-like substance from tumor. The amount of this material obtained from tumor was approximately one-seventh that obtained from muscle.

Received 11/ 1/49.