INHIBITION OF INVASIVENESS AND METASTATIC POTENTIAL OF HUMAN PE-B LYMPHOME CELLS BY INHIBITING UROKINASE EXPRESSIN

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Although I have moved to Okayama University Medical School from Toyama Medical and Pharmaceutical University January 1998, the above research is continuously but slowly progressing. A new antisense plasmid covering 540 bp from the translation starting site of uPA gene was constructed by inserting an antisense-oriented cDNA fragment of urokinase gene into a PCI vector, The plasmid was transferred with a plasmid containing a Hygromycin-resistant gene into RC-K8 human pre B lymphoma cells by the electroporation. The cells were cultured in the RPMI- 1640 culture medium containing Hygromycin for 2 months and a number of Hygromycin-resistant clones were established. Among them, a couple of clones exerted a reduced uPA production, approximately a half of that of parent RC-K8 cells. The invasiveness examined using a Matrigel chamber was not significantly different from that of parent cells. These results were unfortunately very similar to that seen in the previous experiments performed in 1996 and 1997. Obviously I could not have the conclusion at present, because I do not suceeed yet in construction of the RC-K8 cells whose uPA production was completely bloc ed. Now I am trying to get the urokinase-nonproducing RC-K8 cells using two different antisense plasmid-vectors and proceeding the research with much more careful election and subcloning.