[The inhibitory effect of asODNs on the invasion of colorectal cancer cell line CCL229].
To study the inhibitory effect of antisense oligodeoxynucleotide (asODNs) on colrectal cancer cell line CCL229 invasion in vitro. A 15-mer asODNs targeted against the translation start site of UPAR (urokinase-type plasminogen activator receptor) mRNA were introduced into CCL229 cells by lipid-mediated DNA-transfection and the variation of the levels of uPAR mRNA, uPAR antigen expression of the levels of uPAR mRNA, uPAR antigen expression on the cell sruface and invasion properties were observed by reverse transcription-polymerase chain reaction (RT-PCR), flowcytometry(FCM) and aminion invasion assay, the morphological feature of the cell after asODNs treatment was observed by scanning electron microscope(SEM). The results indicate (1) the uPAR/beta-actin ratio was 0.44 +/- 0.02 for the asODNs treated cells, which is significantly lower compared with the control and rONDs treated cells (0.81 +/- 0.01 and 0.750 +/- 0.13 respectively, P < 0.01), (2) the mean fluorescence index of uPAR combined with uPA and the whole uPAR on surface were 0.20 +/- 0.07 and 0.59 +/- 0.09 respectively for asODNs treated cells, which is significantly lower compared with control cells (0.72 +/- 0.12 and 2.21 +/- 0.36 respectively, P < 0.05, P < 0.01); (3) the number of cells migrated the aminion (25 +/- 4, 44 +/- 5 for the control cells) obviously decreased after a-sODNs treatment, (12 +/- 2, 20 +/- 3, P < 0.05); (4) the filopodia and microspikes on the CCL 229 cell surface were decreased after asODNs treatment. The conclusion is that the expression of uPAR on the surface of CCL229 cell surface is responsible for invasity; the inhibitory effect of uPAR as ODNs were highly significant and this method may be of potential clinical interest in gene therapy of cancer.
Jin
Y J
YJ
Institute of Medical Molecular Biology, China Medical University, Key Laboratory of Cell Biology, Ministry of Public Health of China, Shenyang 110001.
Song
J D
JD
D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D009361:Neoplasm Invasiveness; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D014162:Transfection; D014407:Tumor Cells, Cultured
D016376:Oligonucleotides, Antisense; C524898:PLAUR protein, human; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator