Chromosome 17p13.2 transfer reverts transformation phenotypes and Fas-mediated apoptosis in breast epithelial cells.
Transformation of the human breast epithelial cells (HBEC) MCF-10F with the carcinogen benz(a)pyrene (BP) into BP1-E cells resulted in the loss of the chromosome 17 p13.2 locus (D17S796 marker) and formation of colonies in agar-methocel (colony efficiency (CE)), loss of ductulogenic capacity in collagen matrix, and resistance to anti-Fas monoclonal antibody (Mab)-induced apoptosis. For testing the role of that specific region of chromosome 17 in the expression of transformation phenotypes, we transferred chromosome 17 from mouse fibroblast donors to BP1-E cells. Chromosome 11 was used as negative control. After G418 selection, nine clones each were randomly selected from BP1-E-11neo and BP1-E-17neo hybrids, respectively, and tested for the presence of the donor chromosomes by fluorescent in situ hybridization and polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) analyses. Sensitivity to Fas Mab-induced apoptosis and evaluation of transformation phenotype expression were tested in MCF-10F, BP1-E, and nine BP1-E-11neo and BP1-E-17neo clones each. Six BP1-E-17neo clones exhibited a reversion of transformation phenotypes and a dose dependent sensitivity to Fas Mab-induced apoptosis, behaving similarly to MCF-10F cells. All BP1-E-11neo, and three BP1-E-17neo cell clones, like BP1-E cells, retained a high CE, loss of ductulogenic capacity, and were resistant to all Fas Mab doses tested. Genomic analysis revealed that those six BP1-E-17neo clones that were Fas-sensitive and reverted their transformed phenotypes had retained the 17p13.2 (D17S796 marker) region, whereas it was absent in all resistant clones, indicating that the expression of transformation phenotypes and the sensitivity of the cells to Fas-mediated apoptosis were under the control of genes located in this region.
Lareef
Mohamed H
MH
Breast Cancer Research Laboratory, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA.
Tahin
Quivo
Q
Song
Joon
J
Russo
Irma H
IH
Mihaila
Dana
D
Slater
Carolyn M
CM
Balsara
Binaifer
B
Testa
Joseph R
JR
Broccoli
Dominique
D
Grobelny
Jennifer V
JV
Mor
Gil
G
Cuthbert
Andrew
A
Russo
Jose
J
D000818:Animals; D017209:Apoptosis; D001564:Benzo(a)pyrene / Q000633:toxicity; D001940:Breast / Q000166:cytology* / Q000201:enzymology / Q000473:pathology; D002455:Cell Division / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D002478:Cells, Cultured; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics*; D002999:Clone Cells / Q000166:cytology / Q000201:enzymology; D003094:Collagen / Q000378:metabolism; D003114:Colony-Forming Units Assay; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D051379:Mice; D018895:Microsatellite Repeats / Q000235:genetics; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D019098:Telomerase / Q000378:metabolism; D014162:Transfection; D019014:fas Receptor / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D019014:fas Receptor; D001564:Benzo(a)pyrene; D003094:Collagen; D019098:Telomerase