Conversion of high grade lymphoma tumor cell line to intermediate grade with TPA and bryostatin 1 as determined by polypeptide analysis on 2D gel electrophoresis.
A high-resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used for total cellular polypeptide mapping of two established lymphoma cell lines; MANCA, a Burkitt's line representing a high grade lymphoma (HGL) and WSU-NHL (nodular histiocytic lymphoma) representing an intermediate grade lymphoma (IGL). Gels were digitized and analysed with an image scanning computer using the Elsie 4 system. Polypeptide mapping revealed striking similarities in 1100 polypeptide spots in both types. However, three polypeptides were unique to HGL (Molecular mass/isoelectric point (Mr/PI): 39/4.4, 35/5.6, 33/4.8), and one to IGL (95/4.7). In order to investigate the kinetics of expression of these polypeptides, the two cell lines were treated with two protein kinase C (PKC) activators, tumour promoting 12-O tetradecanoylphorbol 13-acetate (TPA) and bryostatin 1. 2D-PAGE of the treated cells revealed that the HGL line loses its unique polypeptides and expresses a new one. The new polypeptide has the same Mr and PI as that unique to the untreated IGL (95/4.7). TPA or bryostatin 1 treatment of the IGL line for 72 h induced no significant changes. Our data show a unidirectional change from HGL to IGL, supporting the clinical notion that HGL is less differentiated than IGL. It also shows the similarity in the mode of action of bryostatin 1 and TPA in inducing these polypeptide changes.
al-Katib
A
A
Division of Hematology and Oncology, Wayne State University, Detroit, MI 48202.
Mohammad
R M
RM
Mohamed
A N
AN
Pettit
G R
GR
Sensenbrenner
L L
LL
D054713:Bryostatins; D002454:Cell Differentiation; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007783:Lactones / Q000494:pharmacology*; D008223:Lymphoma / Q000032:analysis* / Q000235:genetics / Q000473:pathology; D018942:Macrolides; D010455:Peptides / Q000032:analysis*; D010641:Phenotype; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D054713:Bryostatins; D007783:Lactones; D018942:Macrolides; D010455:Peptides; C046785:bryostatin 1; D013755:Tetradecanoylphorbol Acetate