特异性小干扰性RNA降低骨桥蛋白基因表达对肝癌细胞侵袭性的影响  

Knocking down osteopontin expression by specific siRNA reduces the in vitro invasiveness of human hepatocellular carcinoma cells

作　　者：朱晓群[1] 叶青海[1] 雷科峰[1] 陈洁[1] 钦伦秀[1] 

机构地区：[1]复旦大学附属中山医院肝癌研究所肝外科,上海200032

出　　处：《中华肿瘤杂志》2006年第6期404-407,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金(30371378);国家863计划重点项目(2002BA711A02-4);国家杰出青年基金(30325041);教育部跨世纪优秀人才培养计划(教技函2002-48);教育部重大项目(教技司2001-171)

摘　　要：目的应用RNA干扰技术降低细胞内骨桥蛋白的表达水平,研究骨桥蛋白对人肝癌细胞侵袭性的影响。方法体外化学合成骨桥蛋白序列特异性双链RNA,转染高转移肝癌细胞株HCC-LM3,用实时定量PCR和Western blot检测骨桥蛋白的表达。通过生长曲线、克隆形成和Matrigel侵袭实验,观察肝癌细胞恶性生物学行为的改变。结果骨桥蛋白特异性小干扰性RNA (siRNA)转染组的HCC-LM3细胞,骨桥蛋白mRNA水平下降79%,蛋白水平下降81%。体外实验表明,与空白对照组及非特异性siRNA转染组相比,骨桥蛋白特异性siRNA转染组HCC-LM3细胞克隆形成数目下降,穿过人工基底膜的细胞数减少(P＜0．05)。结论降低骨桥蛋白的表达可显著抑制肝癌细胞的侵袭转移能力。Objective To study the effect of osteopontin （OPN） expression down-regulated by RNA interference （ RNAi ） on the invasiveness of hepatocelluar carcinoma cell line HCC-LM3. Methods HCC-LM3 cells were transfected with the chemically synthesized small interfering RNA （siRNA） formulated by lipofectamine 2000. Wild type HCC-LM3 and HCC-LM3 cells transfected with non-specific siRNA served as controls. Real-time PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The malignant phenotypes of transfected HCC-LM3 cells including cellular growth rate, colony formation and Matrigel invasion activities were analyzed. Results Sequence-specific siRNAs targeting OPN suppressed OPN RNA expression by 79% and also decreased OPN protein level by 81% in HCC-LM3 cells. The number of formed colonies and migrating numbers in vitro were decreased in HCC-LM3 cells transfected using sequence-specific siRNAs targeting OPN relative to controls （ P 〈 0. 05 ）. Conclusion This study demonstrated that specific siRNA is able to reduce OPN at both the mRNA and protein levels and significantly diminishes the invasiveness of hepatocellular carcinoma cells.

关 键 词：骨桥蛋白 SIRNA 肝细胞癌 基因表达 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]