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PMID: 8055871 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Electrophoretic analysis of proteins associated with tumor cell invasion.

Electrophoresis ·Vol. 15 ·No. 3-4 ·1994-00-00 ·Pages 454-62

Seftor RE

Abstract

Polyacrylamide gel electrophoresis is an extremely powerful tool for separating and analyzing protein associated with different diseases and has been invaluable in the identification and analysis of proteins associated with characteristics unique to tumor cells. This study presents data demonstrating the application of conventional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and substrate-incorporated SDS-polyacrylamide gel electrophoresis (zymography) to obtain information about the proteins and catalytically active (or activatable) proteases associated with the process of tumor cell invasion using established human melanoma and breast carcinoma cell lines. Conventional SDS-polyacrylamide gel electrophoresis was used to show that cells sequentially selected from a low invasive human melanoma cell line on the basis of their ability to invade in vitro have an increase and/or addition of six unique proteins on their cell surface. In a different application of SDS-polyacrylamide gel electrophoresis, zymography was used to demonstrate that there is an increase in the levels of gelatinase A in the conditioned medium from three differently invasive human melanoma cell lines coincident with their ability to invade in vitro. Furthermore, the conditioned medium from the most invasive melanoma cell line demonstrated the greatest amount of gelatinase B activity. While the conditioned medium from three human breast carcinoma cell lines contained low levels of both gelatinase A and B, one breast cell line also contained activity associated with stromelysin(s) not seen in the melanoma cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH Terms
Autoradiography/methods Cell Line Electrophoresis, Polyacrylamide Gel/methods Endopeptidases/biosynthesis,isolation & purification Gelatinases/biosynthesis,isolation & purification Humans Matrix Metalloproteinase 3 Melanoma/metabolism,pathology Membrane Proteins/biosynthesis,isolation & purification Metalloendopeptidases/biosynthesis,isolation & purification Methionine/metabolism Molecular Weight Neoplasm Invasiveness Neoplasm Proteins/biosynthesis,isolation & purification Plasminogen Activators/biosynthesis,isolation & purification Sulfur Radioisotopes Tumor Cells, Cultured
Chemicals
Membrane Proteins Neoplasm Proteins Sulfur Radioisotopes Methionine Endopeptidases Plasminogen Activators Gelatinases Metalloendopeptidases Matrix Metalloproteinase 3
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Seftor R E
Pediatric Research Institute, Cardinal Glennon Children's Hospital, St. Louis University School of Medicine, MO 63110.
Article Info
Journal
Electrophoresis
Abbr.
Electrophoresis
ISSN
0173-0835
Published
1994-00-00
Pages
454-62
Language
English
Region
Germany
NLM ID
8204476
Subset
IM
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