Inserting chromosome 18 into pancreatic cancer cells switches them to a dormant metastatic phenotype.
We demonstrated previously that restoration of chromosome 18 suppressed growth of pancreatic cancer cells in vitro, as well as that of tumors inoculated into nude mice. We also demonstrated that loss of 18q was associated with poor prognosis. Hence there is the possibility that the 18q arm harbors a gene(s) implicated in tumor progression and/or metastasis. In this study, we evaluated the effect of restoring chromosome 18 on metastasis in a few human pancreatic cancer cell lines with and without inactivation of SMAD4. After microcell-mediated chromosome 18 transfer, hybrid cells showed more than a 10-fold weaker metastatic ability than corresponding parental cells; mice injected with 1.25 x 10(6)/250 micro l hybrid clones via tail vein had less than one-tenth of the number of macroscopic metastases in the lung when compared with the control cells. Microscopic examination confirmed the decrease in the number of metastatic lesions. After inoculation of hybrid cells, more than 80% of the high-power fields showed no micrometastases, contrasting with their abundance after using the parental cells. Hybrid cells restored maspin expression irrespective of SMAD4 status in corresponding parental cells. On the other hand, significantly lower vascular endothelial growth factor and matrix metalloproteinase 2 secretion was observed by measuring levels in the conditioned media (CM); the averages were 22% and 20%, respectively. Angiogenesis assays using in vivo Matrigel plugs demonstrated that less neovascularization was observed in nude mice with hybrid cells than with corresponding parental cells. When cells were treated with CM from hybrids, the migration of human umbilical vascular endothelial cells was decreased, but it was partially restored with anti-vascular endothelial growth factor neutralizing antibody, as compared with CM from parental cells. These data represent the first functional evidence suggesting that chromosome 18q encodes a gene that strongly suppresses metastatic activity, possibly through dormancy.
Department of Molecular Pathology, Tohoku University School of Medicine, Sendai 980-8575, Japan.
Liviu P
LP
Lefter
Makoto
M
Sunamura
Toru
T
Furukawa
Kazunori
K
Takeda
Noriko
N
Kotobuki
Mitsuo
M
Oshimura
Seiki
S
Matsuno
Akira
A
Horii
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement; D002478:Cells, Cultured; D002887:Chromosomes, Human, Pair 18; D003094:Collagen / Q000378:metabolism / Q000494:pharmacology; D018450:Disease Progression; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016147:Genes, Tumor Suppressor; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin / Q000378:metabolism / Q000494:pharmacology; D008168:Lung / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D018895:Microsatellite Repeats; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D010190:Pancreatic Neoplasms / Q000235:genetics*; D010641:Phenotype; D011506:Proteins / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015843:Serpins / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D004338:Drug Combinations; D007797:Laminin; D011506:Proteins; D011509:Proteoglycans; C085344:SERPIN-B5; D015843:Serpins; D042461:Vascular Endothelial Growth Factor A; C059485:matrigel; D003094:Collagen; D020778:Matrix Metalloproteinase 2