The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy

Yoon Hoh Kook, John Adamski, Arthur Zelent, Liliana Ossowski

Research output: Contribution to journalArticle

172 Scopus citations

Abstract

Concomitant expression of urokinase type plasminogen activator (uPA) and its surface receptor (uPAR) has been shown to correlate strongly with a more invasive tumor cell phenotype. A highly malignant human epidermoid carcinoma cell line (HEp3) was transfected with a vector capable of expressing an antisense transcript complementary to 300 bases of the 5' end of uPAR, including the ATG codon. Six stably transfected antisense (AS-2, 3, 5, 9, 10, 12) and eight control clones were characterized. All clones produced high levels of uPA activity. Examination of collagenase production and doubling time showed that all of the clones tested produced similar activities. The antisense clones showed a 20-74% reduction in the uPAR sites; the uPAR mRNA level was also reduced. A test of the invasive ability of all clones in a modified chorioallantoic membrane (CAM) showed that invasiveness of the antisense-inhibited clones was directly proportional to the density of surface uPAR. The AS-2 clone, which expressed the lowest number of uPARs showed a significantly reduced level of invasion. The invasiveness of additional AS-inhibited clones was also reduced. Seven control and four AS-inhibited clones were tested for tumorigenicity on CAMs of chick embryos. Inoculation of control cells produced large tumors, while the AS clones were non-tumorigenic. AS-2 did not produce tumors even if kept in vivo for up to 10 weeks. AS-2 and the Neo-1 control clone produced tumors in the majority of injected mice, but while tumors produced by control cells became palpable within the first week after inoculation, the AS-2 tumors first appeared within 4 weeks of inoculation and were less locally invasive and metastatic. Cells harvested from the AS-2 tumors grown in nude mice all expressed a high level of surface uPAR, indicating a selection for this property under in vivo conditions. Overall these results show that diminished expression of surface uPAR leads to a reduction in invasiveness of tumor cells and an increase in tumor latency. Diminished malignant potential early in the establishment of tumors may shift the balance in favor of the host, leading to reduced tumorigenicity and metastasis.

Original languageEnglish (US)
Pages (from-to)3983-3991
Number of pages9
JournalEMBO Journal
Volume13
Issue number17
DOIs
StatePublished - 1994

Keywords

  • Antisense
  • Invasion
  • Metastasis
  • Plasminogen activator receptor

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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