Defective Myb Function Ablates Cyclin E1 Expression and Perturbs Intestinal Carcinogenesis.
Cyclin E1 is essential for the reentry of quiescent cells into the cell cycle. When hypomorphic mutant Myb mice (Myb(Plt4)) were examined, it was noted that Cyclin E1 (Ccne1) expression was reduced. Furthermore, the induction of Ccne1 in recovering intestinal epithelia following radiation-induced damage was ablated in Myb-mutant mice. These data prompted us to investigate whether Myb directly regulated Ccne1 and to examine whether elevated Myb in colorectal cancer is responsible for Cyclin E1-driven tumor growth. Here, it was found that Myb/MYB and Ccne1/CCNE1 expressions were coupled in both mouse and human adenomas. In addition, the low molecular weight Cyclin E1 was the predominant form in intestinal crypts and adenomatous polyposis coli (Apc)-mutant adenomas. Chromatin immunoprecipitation (ChIP) analysis confirmed that Myb bound directly to the Ccne1 promoter and regulated its endogenous expression. In contrast, Myb(Plt4) served as a dominant-negative factor that inhibited wild-type Myb and this was not apparently compensated for by the transcription factor E2F1 in intestinal epithelial cells. Myb(Plt4/Plt4) mice died prematurely on an Apc(Min/) (+) background associated with hematopoietic defects, including a myelodysplasia; nevertheless, Apc(Min/) (+) mice were protected from intestinal tumorigenesis when crossed to Myb(Plt4/) (+) mice. Knockdown of CCNE1 transcript in murine colorectal cancer cells stabilized chromosome ploidy and decreased tumor formation. These data suggest that Cyclin E1 expression is Myb dependent in normal and transformed intestinal epithelial cells, consistent with a cell-cycle progression and chromosome instability role in cancer.


CONCLUSIONS
This study demonstrates that Myb regulates Cyclin E1 expression in normal gastrointestinal tract epithelial cells and is required during intestinal tumorigenesis.
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia. Latrobe Institute of Molecular Science, Department of Genetics, Latrobe University, Bundoora, Victoria, Australia. Walter and Eliza Hall Institute, Parkville, Victoria, Australia.
Dane
D
Cheasley
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia.
Lloyd
L
Pereira
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia.
Shienny
S
Sampurno
Walter and Eliza Hall Institute, Parkville, Victoria, Australia.
Oliver
O
Sieber
Walter and Eliza Hall Institute, Parkville, Victoria, Australia.
Robert
R
Jorissen
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia. Department of Pathology, University of Melbourne, Melbourne, Victoria, Australia.
Huiling
H
Xu
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia.
Markus
M
Germann
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia. Department of Pathology, University of Melbourne, Melbourne, Victoria, Australia.
Yan
Y
Yuqian
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia. Department of Pathology, University of Melbourne, Melbourne, Victoria, Australia. rob.ramsay@petermac.org.
Robert G
RG
Ramsay
Sir Peter MacCallum Oncology Department, Differentiation and Transcription Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia. University of Melbourne, Melbourne, Victoria, Australia. Department of Pathology, University of Melbourne, Melbourne, Victoria, Australia.
Jordane
J
Malaterre
D000236:Adenoma / Q000378:metabolism; D000818:Animals; D063646:Carcinogenesis / Q000378:metabolism*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D047369:Chromatin Immunoprecipitation; D043171:Chromosomal Instability; D002875:Chromosomes / Q000648:ultrastructure; D015179:Colorectal Neoplasms / Q000378:metabolism*; D019927:Cyclin E / Q000378:metabolism*; D018450:Disease Progression; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006410:Hematopoiesis; D006801:Humans; D047468:Immunoprecipitation; D007413:Intestinal Mucosa / Q000378:metabolism; D007414:Intestinal Neoplasms / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D015513:Oncogene Proteins / Q000378:metabolism*; D020626:Oncogene Proteins v-myb / Q000235:genetics*; D011003:Ploidies; D011401:Promoter Regions, Genetic
C475820:CCNE1 protein, human; D019927:Cyclin E; D015513:Oncogene Proteins; D020626:Oncogene Proteins v-myb; C568314:cyclin E1, mouse