Hyperstable U1snRNA complementary to the K-ras transcripts induces cell death in pancreatic cancer cells.
Bottom Line: In this study, we utilised U1 small nuclear RNA (snRNA) that binds physiologically to the 5' splice site (5'ss) of pre-mRNA, to develop a novel vector system that permits imposed binding of antisense RNA to its target.This revealed that two of the hyperstable U1snRNAs induced cell death after gene transduction, and significantly reduced the number of G418-resistant colonies to less than 10% of the controls.Hyperstable U1snRNA might be a novel approach to express effective antisense RNA in target cells.
Affiliation: Investigative Treatment Division, National Cancer Center Research Institute East, 6-5-1, Kashiwanoha, Kashiwa, Chiba 277-8577, Japan.Show MeSH
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Mentions: The induction of cell death so soon after transfection made it difficult to analyse the underlying mechanism biochemically using the transiently transfected cells. However, since the size of several colonies selected with G418 was apparently smaller in r1A and r3A than in the controls, we speculated that these surviving cells expressed relatively low levels of U1snRNA, which permitted colony formation. Thus, we characterised these clones of PANC-1 cells transfected with r1A and r3A (see Table 1Table 1Summary of biological properties of the stable hyperstable U1snRNA transfectants). Among these clones, two out of six r1A-transfected PANC-1 clones (Pr1A) and three out of five Pr3A clones were quite heterogeneous in morphology, and grew three to four times more slowly than other colonies (Table 1 and Figure 3Figure 3
Affiliation: Investigative Treatment Division, National Cancer Center Research Institute East, 6-5-1, Kashiwanoha, Kashiwa, Chiba 277-8577, Japan.