Phenotypic alterations in Kaposi's sarcoma cells by antisense reduction of perlecan.
Metastasis is a sequence of events including proliferation, migration, adhesion, invasion and subsequent metastatic growth of tumour cells in distant organs. We previously showed that highly metastatic variants of murine melanoma cells express higher levels of the basement membrane proteoglycan perlecan than low or non metastatic variants and expression of an antisense perlecan can reduce metastatic potential. In contrast, antisense expression of perlecan in fibrosarcoma cells was reported to enhance tumorigenesis. To better understand the role of perlecan in angiogenesis we have transfected KS-IMM, an immortalized cell line derived from a human Kaposi s sarcoma, with an antisense perlecan construct and investigated the positive/negative role of perlecan in KS. KS-IMM cells were transfected with either empty vector (neo) or the antisense perlecan construct and clones were isolated. Immuno-blot analysis showed a reduction of perlecan levels in two (AP3 and AP4) isolated clones, in Northern blot analysis endogenous perlecan was undetectable in the AP3 and AP4 clones, while it was present in the neo control clones. AP clones had a reduced migration to HGF in Boyden chambers as compared to neo clones. Proliferation in low serum or serum-free conditions was strongly reduced in the AP clones as compared to the neo control cells. The neotransfected cells showed rapid proliferation in low serum supplemented with HGF and VEGF, while antisense transfected clones showed little response. Finally, AP-trasfected KS-IMM cells had significantly reduced migration to VEGF and HGF with respect to controls. In contrast, when the AP transfected cells were injected in nude mice they paradoxically showed enhanced tumor growth as compared to controls. Our preliminary data indicate that perlecan reduction plays a crucial role on Kaposi s sarcoma cell migration and proliferation in vitro. However, in vivo KS-IMM depleted of perlecan had a growth advantage. A possible hypothesis is that perlecan is necessary for growth of KS-IMM cells in vitro, however its down-regulation might promote angiogenesis through increased angiogenic growth factor diffusion, resulting in enhanced tumor growth in vivo.
Marchisone
C
C
Istituto Nazionale per la Ricerca sul Cancro, Modulo di Progressione Neoplastica, Genova, 16132, Italy.
Del Grosso
F
F
Masiello
L
L
Prat
M
M
Santi
L
L
Noonan
D M
DM
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects; D016895:Culture Media, Serum-Free; D018076:DNA, Complementary / Q000235:genetics; D016228:Endothelial Growth Factors / Q000494:pharmacology; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D006801:Humans; D008222:Lymphokines / Q000494:pharmacology; D051379:Mice; D008819:Mice, Nude; D008954:Models, Biological; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D011509:Proteoglycans / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D017978:Receptors, Growth Factor / Q000502:physiology; D012514:Sarcoma, Kaposi / Q000378:metabolism / Q000473:pathology*; D015398:Signal Transduction; D012983:Soft Tissue Neoplasms / Q000378:metabolism / Q000473:pathology*; D014162:Transfection; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016895:Culture Media, Serum-Free; D018076:DNA, Complementary; D016228:Endothelial Growth Factors; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D017228:Hepatocyte Growth Factor; D008222:Lymphokines; D009363:Neoplasm Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D011509:Proteoglycans; D017978:Receptors, Growth Factor; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; C071927:perlecan