摘要
目的 研究层粘连蛋白受体 (LNR)反义寡核苷酸硫代修饰片段 (AS OD)对人胆管癌QBC939细胞尿激酶型纤维蛋白溶解酶原激酶 (u PA)基因表达的调节。方法 应用逆转录 聚合酶链反应 (RT PCR)分析浓度 12 μmol/L的LNRAS OD在作用 4 8h后对QBC939细胞u PAmRNA表达水平的影响。结果 LNRAS OD能明显抑制人胆管癌细胞表达u PA ,与正常对照相比 ,其u PAmRNA水平下降了 30 0 % (P <0 0 5 )。结论 胆管癌细胞u PA基因表达水平与层粘连蛋白受体的表达水平明显相关 ,抑制层粘连蛋白受体的表达能够使胆管癌细胞u PA基因表达下降 。
Objective To investigate the effect of antisense oligonucleotide (AS OD) of laminin receptor (LNR) on urokinase type plasminogen activator (u PA) expression in human bile duct carcinoma cells Methods Bile duct carcinoma cells of QBC939 line were cultured Compounds of lipofectAMINE and AS OD phosphorothioate of LNR of the concentrations of 3, 6, and 12 μmol/L respectively were prepared and then transfected into the QBC939 cells Sense oligodeoxynucleotide (S OD) of LNR and blank vector were used as controls After culture of 48 hours, flow cytometry was used to detect the protein expression of LNR and the u PA mRNA expression was detected by reverse transcriptase polymerase chain reaction (RT PCR) Cell invasion was examined by culture plate with Transwell microporous film before and after transfection Results Flow cytometry showed down regulation of LNR protein expression in the cells transfected with AS OD dose dependently The LNR protein expression of the cells transfected with AS OD of the concentration of 6 μmol/L was down regulated by over 25% in comparison with those of the blank vector group and S OD group (both P <0 05) RT PCR showed that the u PA mRNA expression of the AS OD 6 μmol/L group was down regulated by 30% in comparison with the 2 control groups The number of migrating cells in the AS OD group was 78±6, significantly lower than those in the S OD group (105±11), and blank vector group (101±7), both P <0 05 Conclusion The expression of laminin receptor in the bile duct carcinoma cells is related with the expression of u PA gene expression Inhibition of LNR expression decreases the u PA gene expression, suggesting that LNR plays an important role in the invasion and metastasis of bile duct carcinoma
出处
《中华医学杂志》
CAS
CSCD
北大核心
2004年第19期1642-1644,共3页
National Medical Journal of China
基金
国家自然科学基金资助项目 (3 9870 83 0 )
