Asbestos induction of extended lifespan in normal human mesothelial cells: interindividual susceptibility and SV40 T antigen.
Normal human mesothelial cells from individual donors were studied for susceptibility to asbestos-induction of apoptosis and generation of an extended lifespan population. Such populations were generated after death of the majority of cells and arose from a subset of mesothelial cultures (4/16) whereas fibroblastic cells (5/5) did not develop extended lifespan populations after asbestos exposure. All mesothelial cultures were examined for the presence of SV40 T antigen to obtain information on (i) the presence of SV40 T antigen expression in normal human mesothelial cells and (ii) the relationship between generation of an extended lifespan population and expression of SV40 T antigen. Immunostaining for SV40 T antigen was positive in 2/38 normal human mesothelial cultures. These cultures also had elevated p53 expression. However, the two isolates expressing SV40 T antigen did not exhibit enhanced proliferative potential or develop an extended lifespan population. Asbestos-generated extended lifespan populations were specifically resistant to asbestos-mediated but not to alpha-Fas-induced apoptosis. Deletion of p16Ink4a was shown in 70% of tumor samples. All mesothelioma cell lines examined showed homozygous deletion of this locus which extended to exon 1beta. Extended lifespan cultures were examined for expression of p16Ink4a to establish whether deletion was an early response to asbestos exposure. During their rapid growth phase, extended lifespan cultures showed decreased expression of p16Ink4a relative to untreated cultures, but methylation was not observed, and p16Ink4a expression became elevated when cells entered culture crisis. These data extend the earlier observation that asbestos can generate extended lifespan populations, providing data on frequency and cell type specificity. In addition, this report shows that generation of such populations does not require expression of SV40 T antigen. Extended lifespan cells could represent a population expressing early changes critical for mesothelioma development. Further study of these populations could identify such changes.
Xu
L
L
Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, MD 20892, USA.
Flynn
B J
BJ
Ungar
S
S
Pass
H I
HI
Linnainmaa
K
K
Mattson
K
K
Gerwin
B I
BI
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects; D001194:Asbestos / Q000009:adverse effects*; D017639:Asbestos, Amosite / Q000009:adverse effects; D002273:Carcinogens / Q000009:adverse effects*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D016922:Cellular Senescence / Q000187:drug effects* / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000235:genetics; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D005796:Genes; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008654:Mesothelioma / Q000378:metabolism / Q000473:pathology; D008745:Methylation; D008875:Middle Aged; D010997:Pleural Neoplasms / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000032:analysis
D000952:Antigens, Polyomavirus Transforming; C493956:CDKN1A protein, human; D002273:Carcinogens; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D016159:Tumor Suppressor Protein p53; D017639:Asbestos, Amosite; D001194:Asbestos