Coordinate suppression of myeloma-specific genes and expression of fibroblast-specific genes in myeloma X fibroblast somatic cell hybrids.
In most instances, fusion of differentiated cell types with fibroblasts has resulted in the extinction of the differentiation-specific traits of the non-fibroblast parental cell. To explore the genetic basis of this phenomenon, we have studied a series of somatic cell hybrids between mouse myeloma and fibroblasts. All the hybrids were adherent having a fibroblast-like phenotype. Molecular analysis revealed that plasma cell specific genes like the productively rearranged Ig genes, the J chain gene and genes for the cell surface markers CD20 and PC1, were extinguished in the hybrids. In contrast, fibroblast specific genes like fibronectin, alpha 2(I) and III collagens, as well as the receptor for fibroblast growth factor (flg), were expressed. Extinction was not due to chromosomal loss or lack of the relevant genes. To learn about the mechanism(s) of this phenomenon we have looked for the presence of positive and negative transcription factors in our hybrids. Expression of the PU.1 transcription factor, a member of the Ets transcription factor family normally expressed in B cells and macrophages, was lost in the cell hybrids. Interestingly, we found that the B-cell-specific Oct-2 transcription factor was still expressed at somewhat variable levels in several of the hybrid cell lines. In contrast, expression of the recently identified octamer coactivator BOB.1/OBF.1 was extinguished in all cell hybrids. This supports a critical role of this transcriptional coactivator for B-cell-specific gene expression. In addition, the Id and HLH462 genes coding for proteins known to repress bHLH transcription factors by formation of heterodimers, were found to be expressed at increased levels in fibroblasts and in the hybrids, indicating that their increased levels might also contribute to the suppression of myeloma-specific genes. Our results show that in myeloma x fibroblast hybrids, the phenotype of the fibroblast is dominant. It is suggested that fibroblasts contain regulatory "master" genes that are responsible for activation of the fibroblast differentiation pathway and suppress differentiation programs of other cell types.
Reich
L
L
Hubert H. Humphrey Center for Experimental Medicine and Cancer Research, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Sharir
H
H
Ber
R
R
Wirth
T
T
Bergman
Y
Y
Laskov
R
R
D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics; D001483:Base Sequence; D003094:Collagen / Q000235:genetics; D004247:DNA / Q000032:analysis; D005347:Fibroblasts / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics; D005786:Gene Expression Regulation; D015972:Gene Expression Regulation, Neoplastic; D005803:Genes, Immunoglobulin / Q000235:genetics; D016259:Genes, myc / Q000235:genetics; D006822:Hybrid Cells / Q000378:metabolism*; D051796:Inhibitor of Differentiation Proteins; D051379:Mice; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000235:genetics*; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*
D000944:Antigens, Differentiation, B-Lymphocyte; D005353:Fibronectins; D051796:Inhibitor of Differentiation Proteins; C490424:Pou2af1 protein, mouse; D011506:Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D014157:Transcription Factors; C067679:Idb3 protein, mouse; D003094:Collagen; D004247:DNA