CAJ | 학술논문

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.
목적 탐토응용소우우RNA(siRNA)침묵골교단백(OPN)기인적표체대인간암세포주침습전이적억제작용. 방법우체외화학합성침대OPN서렬특이성적의련RNA(dsRNA),전염인간암세포주(HCC LM3),용실시정량PCR화Western blot검측OPN mRNA수평화단백표체.통과생장곡선、극륭형성화Matrigel침습실험,관찰간암세포생물학행위적지표.통계학방법채용SPSS11.5연건진행q검험,계수자료용χ<'2>검험. 결과여공백조상비,siRNA특이성전염HCC-LM3세포OPN mRNA수평하강84.7%,단백수반하강81%(P<0.5);세포극륭형성수목하강(1.91개대비5.40개,P<0.01);천과인공기저막적세포수감소(13.5개대비33.4개,P<0.05),음성대조조적OPN mRNA여공백대조조차이무통계학의의(29.7개대비33.4개,P>0.05).결론 침묵OPN기인표체-구J재체외조알간암침습.
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.