Immortalization and transformation of human mammary epithelial cells by a tumor-derived Myc mutant.
The Myc transcription factor is commonly dysregulated in many human cancers, including breast carcinomas. However, the precise role of Myc in the initiation and maintenance of malignancy is unclear. In this study we compared the ability of wild-type Myc (wt Myc) or Myc phosphorylation deficient mutants (T58A, S62A or T58A/S62A) to immortalize and transform human mammary epithelial cells (HMECs). All Myc constructs promoted cellular immortalization. As previously reported in other cells, the Myc T58A mutant tempered apoptotic responses and increased Myc protein stability in HMEC cells. More importantly, we now show that HMECs overexpressing the Myc T58A mutant acquire a unique cellular phenotype characterized by cell aggregation, detachment from the substrate and growth in liquid suspension. Coincident with these changes, the cells become anchorage-independent for growth in agarose. Previous studies have shown that wt Myc can collaborate with hTERT in inducing HMEC anchorage-independent growth. We have verified this observation and further shown that Myc T58A was a stronger facilitator of such co-transformation. Thus, our findings indicate that differences in Myc protein phosphorylation modulate its biological activity in human breast epithelial cells and specifically that the T58A mutation can facilitate both cellular immortalization and transformation. Finally, we used the isogenic cell lines generated in this study to identify a subset of genes whose expression is greatly altered during the transition from the immortal to the anchorage-independent states.
Thibodeaux
Clare A
CA
Department of Pathology, Georgetown University Medical School, NW, Washington, DC 20057, USA.
Liu
Xuefeng
X
Disbrow
Gary L
GL
Zhang
Yiyu
Y
Rone
Janice D
JD
Haddad
Bassem R
BR
Schlegel
Richard
R
D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics*; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D004847:Epithelial Cells / Q000473:pathology* / Q000502:physiology; D005260:Female; D015870:Gene Expression; D016259:Genes, myc; D006367:HeLa Cells; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D014161:Transduction, Genetic