Inhibitory effect of cathepsin L antisense nucleotide on biological behavior of human highly metastatic osteosarcoma cell

Objective To set up sense/antisense eukaryotic expression vectors for human cathepsin L(CATL) gene,and to study the biological effects on human highly metastatic osteosarcoma cell line MG-63A1 after transfection.Methods A 1001bp cDNA fragment was amplified by RT-PCR method from total RNA of human osteosarcoma tissue.The fragment was cloned into pcDNA 3.0 plasmid with sense/antisense direction respectively.The recombinant plasmids were confirmed by double digestion with restriction endonucleases BamHⅠ and XbaⅠ and DNA sequencing.The recombinant plasmids were transfected into MG-63A1 cells.Changes of cell proliferation,cell invasion ability and tumorigenecity and spontaneous metastasis in nude mice were detected.Results The sense/antisense recombinant eukaryotic expression vectors were obtained.Sequence analysis further confirmed that the sense recombinent plasmid contained a fragment which is identical to the cDNA sequence of CATL gene,while antisense recombinent plasmid contained a fragment which is in opposite orientation to the cDNA sequence of CATL gene.The vivo/vitro invasion ability was remarkably decreased after antisense transfection compared with that of control cells.Conclusions The vivo/vitro invasion ability of osteosarcoma cells can be remarkably inhibited by antisense CATL.