反义表达人组织蛋白酶L对骨肉瘤细胞侵袭特性的影响

Cathepsin L antisense cDNA in a human osteosarcoma cell line:effects on the invasive phenotype

目的: 构建人组织蛋白酶L (cathepsinL, CATL) 基因的正、反义真核表达载体, 观察其转染后对人骨肉瘤细胞MG-63侵袭特性的影响。方法: 用基因重组技术构建人组织蛋白酶L正、反义真核表达载体, 转染人骨肉瘤细胞MG-63, RT-PCR和Westernblot检测CATL的mRNA和蛋白质的表达水平, 并对转染前后骨肉瘤细胞的侵袭能力进行检测。结果: 正、反义真核表达载体成功构建并转染入MG-63细胞中。反义载体转染后的细胞CATL的mRNA和蛋白质的表达水平下降, 体外侵袭实验表明细胞的侵袭力下降。结论: 反义CATL基因的转染使骨肉瘤细胞CATL的分泌水平下降, 细胞的体外侵袭力下降。

Objective:To obtain sense/antisense eukaryotic expression vectors for human cathepsin L gene,and study the biological effects on human osteosarcoma cell line MG-63 after transfection.Method:Cathepsin L gene sense/antisense eukaryotic expression vectors were constructed with recombinant technology and transfected into the human osteosarcoma cell line MG-63.The expression of cathepsin L gene mRNA was examined with RT-PCR and the expression of cathepsin L was examined with Western blot.The invasion ability of the human osteosarcoma cell line MG-63 after transfection was detected.Result: The sense/antisense recombinant eukaryotic expression vectors for cathepsin L were successfully constructed and transfected into MG-63 cell.The invasion ability was remarkably decreased after antisense transfection compared with that of control cells. Conclusion: Antisense cathepsin L gene can significantly inhibit the expression of cathepsin L mRNA and protein,and down regulate MG-63 cell's invasion ability.