骨桥蛋白基因沉默对肝癌侵袭转移的抑制作用

目的 探讨应用小于扰RNA(siRNA)沉默骨桥蛋白(OPN)基因的表达对人肝癌细胞株侵袭转移的抑制作用. 方法于体外化学合成针对OPN序列特异性的舣链RNA(dsRNA),转染人肝癌细胞株(HCC LM3),用实时定量PCR和Western blot检测OPN mRNA水平和蛋白表达.通过生长曲线、克隆形成和Matrigel侵袭实验,观察肝癌细胞生物学行为的指标.统计学方法采用SPSS11.5软件进行q检验,计数资料用χ<'2>检验. 结果与空白组相比,siRNA特异性转染HCC-LM3细胞OPN mRNA水平下降84.7%,蛋白水半下降81%(P<0.5);细胞克隆形成数目下降(1.91个对比5.40个,P<0.01);穿过人工基底膜的细胞数减少(13.5个对比33.4个,P<0.05),阴性对照组的OPN mRNA与空白对照组差异无统计学意义(29.7个对比33.4个,P>0.05).结论 沉默OPN基因表达-口J在体外阻遏肝癌侵袭.

Interference of osteopontin expression inhibits the invasion and metastasis of human hepatocellular carcinoma cell lines

Objective To investigate the effect of osteopontin (OPN) on the invasion and metastasis of human hapatocellular carcinoma (HCC). Methods HCC cell lines (HCC-LM3) were transfected with the chemically synthesized small interfering RNA (siRNA). Real-time PCR and Western blot were used to quan-tify the mRNA and OPN protein levels. The malignant phenotypes including cellular growth, colony forma-tion and invasion capability of the HCC cells were analyzed. Results The OPN mRNA and proteins levels were decreased by 75% and 80% in OPN siRNA treated cells. Colony formation and migratory capability were reduced in OPN siRNA treated cells (P < 0.05). Conclusion The specific siRNA is able to reduce the OPN expression at both the mRNA and protein levels and significantly inhibits the invasiveness of HCC cells.