目的研究层黏素受体(LNR)反义硫代磷酸寡核苷酸片段(AsODN)对人胆管癌QBC939细胞基质蛋白水解酶-2(MMP-2)、基质蛋白水解酶-9(MMP-9)以及尿激酶型纤维蛋白溶解酶原激酶(u—PA)蛋白表达的影响。方法应用Western blotting分析浓度12/μmol/L的LNR AsODN作用48h后,QBC939细胞MMP-2、MMP-9以及u—PA蛋白表达水平。结果LNR AsODN能明显抑制人胆管癌细胞表达MMP-2、MMP-9以及u-PA,与对照组相比,MMP-2、MMP-9、u—PA蛋白表达下降分别为21.8%、16.5%和9.0%(P〈0.05)。结论胆管癌细胞QBC939蛋白水解酶MMP-2、MMP-9以及u—PA基因表达水平与层黏素受体的表达水平明显相关,LNRAsODN表达能够使胆管癌细胞蛋白水解酶基因表达下降,提示层黏素受体在介导胆管癌转移中发挥重要作用。
Objective To investigate the effect of antisense oligodeoxynucleotide (AsODN) of laminin receptor (LNR) on expressions of matrix metalloproteinases-2 (MMP-2), MMP-9 and u-PA in human cholangiocarcinoma cells. Methods Protein expressions of MMP-2, MMP-9 and u-PA of cell line QBC939 were analyzed by using Western blotting after lipofectAMINE and AsODN phosphorothioate of LNR at concentrations of 12 μmol/L were cultured for 48 hours. Results Western blotting showed that AsODN could significantly down-regulate protein expressions of MMP-2, MMP-9 and u-Pap, with decrease rate of 21.8 %, 16.5 % and 9.0 % respectively ( P 〈0.05) in comparison with the control group. Conclusions Gene expressions of MMP-2, MMP-9 and u-PA are markedly related to expression of laminin receptor in cholangioeareinoma cells and can be decreased by expression of LNR AsODN, suggesting an important role of laminin receptor in the invasion and metastasis of cholangiocarcinoma.