Suppression of tumorigenicity in the human prostate cancer cell line M12 via microcell-mediated restoration of chromosome 19.
Previously we immortalized human, nontransformed prostate epithelial cells with SV40 large T-antigen (SV40TAg) and derived increasingly aggressive sublines from the immortalized line. The progression of the tumorigenic sublines to metastatic capacity was accompanied by the formation of an unbalanced translocation between chromosomes 16 and 19, resulting in loss of 19p and proximal 19q. To test whether the tumorigenic and/or metastatic phenotype was causally related to this genetic alteration, we restored a neo-tagged human chromosome 19 to M12 cells by microcell-mediated transfer and assessed their growth. In vitro, the resultant hybrids grew more slowly in monolayer culture and showed a significant reduction in anchorage-independent growth as compared to M12neo controls. In vivo, all mice (13/13) injected subcutaneously (SC) with control M12neo cells developed tumors after 9-15 days. In contrast, 9/15 mice injected SC with microcell-transferred chromosome 19 hybrid cells failed to form tumors, with 6/15 producing very small tumors after 120 days. Analysis of three of these six tumors showed consistent, new chromosomal changes. Furthermore, in one of the tumors, loss of a chromosome 19 was noted in 40% of the cells. After intraprostatic injections of the hybrid cells, only 2/7 mice developed microscopic tumors, with no metastases. These data suggest the presence of a gene or genes on chromosome 19 that function to suppress growth.
Astbury
C
C
Department of Human Genetics, Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia, USA.
Jackson-Cook
C K
CK
Culp
S H
SH
Paisley
T E
TE
Ware
J L
JL
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002455:Cell Division; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics* / Q000378:metabolism*; D003114:Colony-Forming Units Assay; D020732:Cytogenetic Analysis; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D018014:Gene Transfer Techniques; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D007279:Injections, Subcutaneous; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D011471:Prostatic Neoplasms / Q000209:etiology* / Q000235:genetics*; D013489:Suppression, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism / Q000637:transplantation