Elevated in vitro translation of a 25-kDa protein in renal cell carcinoma.
As a first step in understanding the changes in protein synthesis that occur in renal cell carcinoma, we have prepared poly(A)+ RNA from surgically removed tumors and from their normal tissue counterpart. These RNAs were then translated in vitro in the rabbit reticulocyte lysate system and the synthesized labeled polypeptides were separated by one- and two-dimensional gel electrophoresis. A major 25-kDa primary translation product was observed with all renal cell carcinomas. The synthesis of this protein was barely detectable with the RNA from normal tissue adjacent to the tumor. To determine if this protein could be further processed (removal of signal peptide and (or) core glycosylation), canine pancreatic microsomal membranes were added to the system. This addition resulted in the formation of a vertical row of three additional spots, with the same isoelectric point as the primary translation product and with molecular masses ranging from 27 to 31 kDa. The 31-kDa protein was retained on Concanavalin A. After digestion with endoglycosidase H, it was no longer visible on sodium dodecyl sulfate gels and a new 27-kDa band was generated suggesting that the mature protein was indeed a glycoprotein. Future experiments will be aimed at identifying this protein and examining its potential value as a marker of renal cell carcinoma.
Paradis
G
G
Laboratory of Hormonal Bioregulation, Laval University Hospital Research Centre, Ste-Foy, Que., Canada.
Gaudreau
J
J
Frenette
G
G
Thabet
M
M
Tremblay
R R
RR
Dubé
J Y
JY
D002292:Carcinoma, Renal Cell / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D066298:In Vitro Techniques; D007680:Kidney Neoplasms / Q000235:genetics*; D009363:Neoplasm Proteins / Q000378:metabolism*; D014176:Protein Biosynthesis; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism
D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012313:RNA