Amplification of Ki-ras and elevation of MAP kinase activity during mammary tumor progression in C3(1)/SV40 Tag transgenic mice.
We have previously documented that transgenic mice expressing SV40 Tag regulated by the rat prostatic steroid-binding protein C3(1) 5'-flanking region display multistage mammary tumorigenesis. To delineate genetic changes associated with mammary tumor progression, comparative genomic hybridization (CGH) was performed. CGH revealed a consistent gain of the telomeric region of chromosome 6. This region contains the Ki-ras proto-oncogene. Analyses of genomic DNA by Southern blot demonstrated up to 40-fold amplification of the Ki-ras gene. Ki-ras amplification was detected in 12, 46 and 68% of tumors from 4, 5 and 6 month old mice, respectively, whereas no amplifications were found in any preneoplastic mammary tissues. Tumors bearing Ki-ras gene amplification exhibited high levels of Ki-ras RNA and protein. The over-expressed Ki-Ras protein in these tumors appeared functionally active as indicated by the elevated MAP kinase activity. These data demonstrate that while Ki-ras amplification might not be an early event, there is a strong association between Ki-ras amplification and over-expression and mammary tumor progression in this model. This study also shows that CGH is a powerful and useful technique for identifying chromosomal copy number changes during tumor progression, and that this model may provide a predictable in vivo system for studying gene amplification.
Liu
M L
ML
Laboratory of Cell Regulation and Carcinogenesis, Division of Basic Science, National Cancer Institute, Bethesda, Maryland 20892, USA.
Von Lintig
F C
FC
Liyanage
M
M
Shibata
M A
MA
Jorcyk
C L
CL
Ried
T
T
Boss
G R
GR
Green
J E
JE
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D018450:Disease Progression; D005784:Gene Amplification; D011905:Genes, ras / Q000235:genetics*; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D015674:Mammary Neoplasms, Animal / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D012333:RNA, Messenger / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D018631:ras Proteins / Q000378:metabolism
D017871:Calcium-Calmodulin-Dependent Protein Kinases; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D018631:ras Proteins