Identification of conditioned medium proteins from human cancer cells adapted to serum-free conditions.
Recent studies have shown that human cancer cell lines can be adapted to grow in serum-free, unsupplemented RPMI-1640 (RO) medium. We have developed similar techniques to rapidly identify proteins of interest in serum-free conditioned medium (CM) of human lung cancer cell lines. Classic and variant small cell lung cancer (SCLC) lines were adapted to growth in RO medium. CM from each line was concentrated and fractionated on an anion-exchange column of a fast protein liquid chromatography system. Concentrates of each fraction were loaded onto lanes of minigels of an automated electrophoresis system. Analysis of the chromatograms reveals peaks seen only in CM of the classic SCLC lines. Electrophoretic analysis of the fractions containing these peaks reveal protein bands distinguishing between the subtypes of human SCLC. One protein was purified to homogeneity with subsequent reversed-phase chromatography and identified by protein microsequencing as histone H2B. These automated techniques have general use in the rapid identification of CM proteins associated with the differentiation or progression of the many types of neoplastic cells which can be adapted to growth in RO medium.
Doyle
L A
LA
University of Maryland Cancer Center, Baltimore 21201.
Goldstein
L H
LH
Clingroth
C J
CJ
Cuttitta
F
F
D000595:Amino Acid Sequence; D001322:Autoanalysis; D018288:Carcinoma, Small Cell / Q000737:chemistry* / Q000648:ultrastructure; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000737:chemistry; D008175:Lung Neoplasms / Q000737:chemistry* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis*; D012834:Silver; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
D006657:Histones; D009363:Neoplasm Proteins; D012834:Silver