Translation of messenger RNA from a renal tumor into a product with the biological properties of erythropoietin

• Published in: Experimental hematology Vol. 13; no. 1; p. 23
• Main Authors: Saito, T, Saito, K, Trent, D J, Draganac, P S, Andrews, R B, Farkas, W R, Dunn, C D, Etkin, L D, Lange, R D
• Format: Journal Article
• Language: English
• Published: Netherlands 01.01.1985
• Subjects: Adenocarcinoma - genetics; Adenocarcinoma - metabolism; Animals; Cells, Cultured; Erythropoietin - biosynthesis; Erythropoietin - isolation & purification; Female; Fetus; Humans; Kidney Neoplasms - genetics; Kidney Neoplasms - metabolism; Liver; Mice; Mice, Nude; Neoplasm Transplantation; Oocytes - metabolism; Poly A - isolation & purification; Polycythemia - metabolism; Protein Biosynthesis; RNA, Messenger - metabolism; Xenopus laevis
• ISSN: 0301-472X; 1873-2399

The renal tumor RCC-3-JCK, when transplanted into immunodeficient mice, caused an erythrocytic polycythemia. When grown in culture, the tumor cells secreted a substance into the culture medium that chromatographed by size-exclusion high-performance liquid chromatography similarly to purified human erythropoietin (Ep) and was positive when assayed for Ep by its ability to stimulate erythropoiesis in fetal mouse liver cells (the FMLC assay). The poly(A) + RNA was extracted from the tumor cells and injected into Xenopus oocytes, inducing the appearance of Ep(FMLC) in the oocyte culture medium. Both the tumor cells and oocyte culture media were fractionated by size-exclusion high-performance liquid chromatography, and two fractions with Ep(FMLC) activity were found in the tumor-cell culture medium. Three active fractions were found in the medium from the mRNA-injected oocytes. The largest component from both culture media had the same elution time as a human standard (Ep). The poly(A) + RNA was fractionated by sucrose density-gradient centrifugation and the 8S and 10S fractions were found to induce Ep(FMLC) synthesis when they were injected into the oocytes. We conclude that poly(A) + RNA isolated from the Ep-producing tumor RCC-3-JCK included mRNA for Ep and that the Ep was a translational product of Xenopus oocytes injected with this mRNA.