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Analysis of proviral integration in human mammary epithelial cell lines immortalized by retroviral infection with a temperature‐sensitive SV40 T‐antigen construct
Author(s) -
Stamps Alasdair C.,
Davies Susan C.,
Burman Jacqueline,
O'Hare Michael J.
Publication year - 1994
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910570616
Subject(s) - biology , retrovirus , cell culture , clone (java method) , immortalised cell line , provirus , antigen , progenitor cell , virology , genetics , gene , stem cell , genome
A panel of eight conditionally immortal lines derived by infection of human breast epithelial cells with an amphotropic retrovirus transducing a ts mutant of SV40 large T‐antigen was analyzed with respect to individual retroviral integration patterns. Each line contained multiple integration sites which were clonal and stable over extended passage. Similar integration patterns were observed between individual lines arising separately from the same stock of pre‐immortal cells, suggesting a common progenitor. Retroviral integration analysis of preimmortal cells at different stages of pre‐crisis growth showed changes indicative of a progressive transition from polyclonality to clonality as the cells approached crisis. Each of the immortal lines contained a sub‐set of the integration sites of their pre‐immortal progenitors, with individual combinations and copy numbers of sites. Since all the cell lines appeared to originate from single foci in separate flasks, it is likely that each set arose from a common clone of pre‐immortal cells as the result of separate genetic events. There was no evidence from this analysis to suggest that specific integration sites played any part either in the selection of pre‐crisis clones or in the subsequent establishment of immortal lines. © 1994 Wiley‐Liss, Inc.