Expression of a 46 kDa protein in human pancreatic tumors and its possible relationship with the bile salt-dependent lipase.
Bile salt-dependent lipase (BSDL), an enzyme normally found in human pancreatic secretions is a 100 kDa glycoprotein. A BSDL-specific 477 bp cDNA probe was prepared by performing polymerase chain reaction experiments. This cDNA was used to probe mRNAs extracted from human pancreatic tissue and tumoral cell lines. Two mRNAs were detected in normal human pancreas at 2.2 and 1.3 to 1.5 kb. In human pancreatic tumoral cells, mRNAs encoding for the BSDL were detected using in situ hybridization, and proteins with an M(r) of 46,000 to 48,000 were translated into an in vitro system using mRNAs extracted from these cells. Using an immunoprecipitation procedure, we observed here that the specific BSDL polyclonal antibodies recognized three proteins of 100 +/- 5 (p100), 46 +/- 2 (p46) and 22.7 +/- 1.2 (p23) kDa, respectively in the soluble extracts of normal adult human pancreas. The p100 protein was probably the glycosylated product resulting from the translation of the 2.2 kb transcript. The p46 protein, which electrophoresed as a doublet was the main component immunoprecipitated from extract of a differentiated human pancreatic adenocarcinoma as well as from the extracts of two pancreatic cell lines, BxPC-3 and SOJ-6. In addition, the p46 immunoform of the BSDL was detected in cell-free medium from SOJ-6 cell line and its expression was found to be correlated with the secretion of an esterolytic activity on 4-nitrophenyl caproate, whereas the BxPC-3 cell line neither secreted the p46 nor showed any esterolytic activity on this substrate. The p46 may be either a short variant of BSDL resulting from the translation of the 1.3 to 1.5 kb transcript or a protein structurally related to the enzyme. The p46 doublet immunoform was detected in the human pancreatic secretion.
Roudani
S
S
INSERM U 260, Faculté de Médecine-Timone, Marseille/France.
Pasqualini
E
E
Margotat
A
A
Gastaldi
M
M
Sbarra
V
V
Malezet-Desmoulin
C
C
Lombardo
D
D
D001483:Base Sequence; D015152:Blotting, Northern; D004952:Esters; D006801:Humans; D017403:In Situ Hybridization; D008049:Lipase / Q000032:analysis*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis* / Q000096:biosynthesis; D010190:Pancreatic Neoplasms / Q000737:chemistry* / Q000201:enzymology; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D002787:Sterol Esterase; D014407:Tumor Cells, Cultured
D004952:Esters; D009363:Neoplasm Proteins; C054784:bile salt-stimulated lipase; D002787:Sterol Esterase; D008049:Lipase