Tissue plasminogen activator is required for the growth, invasion, and angiogenesis of pancreatic tumor cells

Background & Aims: Overexpression of tissue-type plasminogen activator (t-PA) in exocrine pancreatic tumors might be a determinant of the aggressive biological behavior of these tumors. Methods: Endogenous t-PA production was suppressed by antisense oligonucleotides or transcripts in CAPAN-1 and RWP-1 cell lines. Reciprocally, the t-PA non-expressing BxPC-3 and PANC-1 cells were stably transfected to overexpress t-PA. Recombinant t-PA and chemical inhibitors were also used on these cells. Clones were assayed for invasion and growth in vitro and in vivo. Results: In vitro, specific inhibition of t-PA expression or activity significantly inhibited the proliferation of t-PA-producing RWP-1, CAPAN-1, and SK-PC-1 cells. Antisense constructs were used to generate RWP-1. clones stably suppressed for t-PA expression (AS clones). These clones had a significantly reduced invasion and proliferation on plastic and in soft agar. The addition of recombinant t-PA rescued the growth of the AS clones to parental levels and was mitogenic for other independent pancreas cell lines. This effect did not require plasmin activity. In athymic mice, RWP-1 AS clones produced tumors fivefold smaller than control clones. AS tumors contained a significantly reduced number of Ki67-positive nuclei, fewer mitotic cells, and a remarkably reduced angiogenic network. Finally, the generation of tetracycline-repressed t-PA transfectants in PANC-1 cells confirmed the activity of t-PA in invasion and proliferation in vitro and in vivo. Conclusions: t-PA, in addition to its known role in invasion, plays other critical roles in pancreas tumor progression, stimulating cancer cell proliferation and tumor-associated angiogenesis.