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Two‐dimensional electrophoretic analysis of down‐regulated proteins in human fibroblasts immortalized by treatment with either 4‐nitroquinoline 1‐oxide or 60 Co gamma rays
Author(s) -
Kondo Tadashi,
Mihara Koichiro,
Inoue Yusuke,
Iijima Mikio,
Namba Masayoshi
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501601177
Subject(s) - immortalised cell line , cell culture , hela , fibroblast , microbiology and biotechnology , isoelectric focusing , gel electrophoresis , polyacrylamide gel electrophoresis , cell , isoelectric point , biology , 4 nitroquinoline 1 oxide , chemistry , biochemistry , cancer , genetics , carcinogenesis , enzyme
Cellular proteins were examined by two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) in order to determine the mechanisms of cellular aging and immortalization of human fibroblasts. A total of 10 cell lines were studied, four of which were immortalized. OUMS‐24F and SUSM‐1 cells were immortalized by repeated treatment with 4‐nitroquinoline 1‐oxide (4NQO), KMST‐6 cells were immortalized by intermittent exposure to 60 Co‐gamma rays, and IMR‐90/SV40 cells were immortalized with simian virus 40 (SV40). None of these immortalized cells were tumorigenic. Four normal human fibroblast cell lines, OUMS‐24, AD 387, KMS‐6 and IMR‐90, and two human cervical cancer cell lines, HeLa and A‐431, were also studied. Applying 2‐D PAGE, the down‐regulation of the identical proteins was observed in the four immortalized cell lines and in the two human cancer cell lines. Their molecular masses were about 80 kDa and the isoelectric points ranged between 5.5 and 6.3. We could not find any information on our proteins from the recent protein data bases. Further characterization of the proteins indicated that they might be phosphorylated cytoskeletal proteins.