Expression of beta-actin during progression of mouse skin tumors.
Recombinant DNA techniques have been employed to isolate sequences that are over-expressed during multi-stage carcinogenesis in the mouse skin. One of these six cDNA sequences, mal-4, detected 1.9-kb transcripts that were expressed at levels 10-fold higher in squamous cell carcinomas (SCCs) in comparison to normal epidermis. A full-length cDNA for mal-4 was obtained from a lambda gt10 cDNA library made from an SCC-producing cell line, PDVC57. DNA sequencing of 1300 out of 1900 bp of the mal-4 cDNA and searching of Genbank showed greater than 95% DNA sequence similarity to mouse beta-actin over the entire cDNA and greater than 98% similarity over the 3' untranslated region, which is unique to the various isoforms of actins. Southern analysis showed no amplification or rearrangement of the beta-actin gene had occurred during tumor progression. The RNA:RNA hybrid protection assay was utilized to screen for the expression of mutated beta-actin(s) in mouse skin tumors. No evidence for mutation was obtained in any of the tumors examined. Fluorescence microscopy of tumor sections stained with rhodamine-conjugated phalloidin showed a peripheral pattern of F-actin localization with no gross differences between papillomas and carcinomas. Approximately equal amounts of beta-actin were observed following two-dimensional gel electrophoresis of proteins extracted from normal epidermis, papillomas or SCCs, indicating that the over-expression of beta-actin RNA in SCCs did not result in an increased steady-state level of beta-actin protein. These results suggest that alterations in actin metabolism accompany tumor progression.
Ostrowski
L E
LE
Radiation Oncology Department, University of Arizona Health Sciences Center, Tucson 85724.
Krieg
P
P
Finch
J
J
Cress
A E
AE
Nagle
R B
RB
Bowden
G T
GT
D000199:Actins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002273:Carcinogens; D002294:Carcinoma, Squamous Cell / Q000139:chemically induced / Q000235:genetics*; D003001:Cloning, Molecular; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D012867:Skin / Q000378:metabolism; D012878:Skin Neoplasms / Q000139:chemically induced / Q000235:genetics*
D000199:Actins; D002273:Carcinogens