Stable expression of antisense urokinase mRNA inhibits the proliferation and invasion of human hepatocellular carcinoma cells

• Published in: Cancer Gene Therapy 2003-02, Vol.10 (2), p.112-120
• Main Authors: Tavian, Daniela, Salvi, Alessandro, De Petro, Giuseppina, Barlati, Sergio
• Format: Article
• Language: English
• Subjects: Antisense DNA; Antisense RNA; Biomedical and Life Sciences; Biomedicine; Biopsy; Cancer cells; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Carcinoma, Hepatocellular - therapy; Cell Division - drug effects; Cell Division - genetics; Cell Movement - genetics; Cell proliferation; Enzymatic activity; Fluorescent Antibody Technique; Gene Expression; Gene Therapy; Genetic aspects; Genetic Therapy - methods; Health aspects; Hepatocellular carcinoma; Hepatoma; Humans; Immunoblotting; Invasiveness; Liver cancer; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Liver Neoplasms - therapy; Neoplasm Invasiveness; original-article; Polymerase Chain Reaction - methods; Prevention; Properties; Receptors, Cell Surface - genetics; Receptors, Cell Surface - metabolism; Receptors, Urokinase Plasminogen Activator; RNA, Antisense - biosynthesis; RNA, Antisense - genetics; RNA, Antisense - pharmacology; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; RNA, Messenger - pharmacology; Transfection; Tumor Cells, Cultured; Tumors; U-Plasminogen activator; Urokinase; Urokinase-Type Plasminogen Activator - antagonists & inhibitors; Urokinase-Type Plasminogen Activator - genetics; Urokinase-Type Plasminogen Activator - metabolism
• ISSN: 0929-1903; 1476-5381; 1476-5500
• DOI: 10.1038/sj.cgt.7700533

Urokinase-type plasminogen activator (u-PA) plays a key role in malignant tumor behavior. We have previously shown that the expression of high levels of u-PA mRNA in human hepatocellular carcinoma (HCC) biopsies was inversely correlated with the survival of the patients. In order to evaluate the involvement of u-PA in the invasive and infiltrating properties of HCC cells, the SKHep1C3 cell line was stably transfected with an expression vector containing the 5′ portion (257 bp) of u-PA cDNA in the antisense orientation. u-PA mRNA expression and its protein level and enzymatic activity were specifically inhibited in the antisense transfectants. A comparable inhibition of the u-PA receptor (u-PAR) mRNA and protein was also evidenced in the antisense transfected cells compared with the control ones. At the functional level, the SKHep1C3-AS cells showed a significant reduction in proliferation, Matrigel invasion, and motility assays compared to parental and vector-alone cells. These results indicate that u-PA is an essential factor in the growth and invasiveness of human hepatocarcinoma cells. Antisense u-PA strategy might be a potential approach to reduce tumor growth as well as the invasive capacity of the malignant cells in HCC.