摘要
目的:探讨RNA干扰(RNA interference,RNAi)沉默促肝细胞再生磷酸酶3(proteins in regenerating liver cell-3,PRL-3)基因对大肠癌细胞侵袭的影响.方法:应用PRL-3基因小干扰RNA(small interfering RNA,siRNA)转染处理大肠癌细胞系HCT116后,采用荧光实时定量PCR方法检测PRL-3基因mRNA水平,分别采用软琼脂集落培养实验和Boyden小室模型实验检测癌细胞的锚着不依赖性增殖和侵袭能力.其次将转染48h的细胞接种裸鼠,观察对癌细胞体内侵袭的影响.结果:与两对照组比较,siRNA组PRL-3 mRNA水平明显降低,且呈浓度和时间依赖性(P<0.05).体外实验发现:与对照组比较,PRL-3 siRNA转染组软琼脂集落形成数和穿过滤膜的癌细胞均呈剂量依赖性减少(P<0.05,P<0.05).体内实验发现:空白对照组和空载对照组有较多癌细胞侵袭癌肿组织周围的横纹肌,并侵犯血管,而siRNA转染组未见这些现象.结论:PRL-3在大肠癌细胞侵袭中起着重要的作用,采用PRL-3 siRNA转染可抑制大肠癌细胞侵袭。
AIM： To investigate the effects of RNA interference-induced gene silence of proteins in regenerating liver cell-3 （PRL-3） on the invasion of human colon cancer cells. METHODS： Colon cancer cell line HCT116 was transfected by PRL-3 small interfering RNA （siRNA）, and the mRNA of PRL-3 was determined by fluorescence quantitative real-time polymerase chain reation （FRQ-PCR） assay. The anchorage-independent growth was examined using colony formation in soft agar, and invasion ability was evaluated by Boyden chamber model. RESULTS： Compared with that in controlgroups, the level mRNA of PRL-3 in cancer cells transfected with PRL-3 siRNA was inhibited in a concentration- and time-dependent manner （P 〈 0.05）. In vitro experiment showed that tranfection of colon cancer cells with PRL-3 siRNA resulted in a significant reduction of both colony formation and cell invasion in a concentration- and time-dependent manner, respectively （P 〈 0.05, P 〈 0.05）. In vivo experiment showed that there were lots of cancer cells invading blood vessel and striped muscle around carcinoma tissue in two control groups, but such phenomenon did not occur in siRNA-transfected groups. CONCLUSION： PRL-3 may play an important role in the invasion of human colon cancer cells, and RNAi-induced PRL-3 down-regulation can inhibit cell invasion ability.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第7期767-770,共4页
World Chinese Journal of Digestology
基金
镇江市科技计划基金资助项目
No.SH2006019~~
关键词
大肠肿瘤
促肝细胞再生磷酸酶3
侵袭
RNA干扰
小干扰RNA
荧光实时定量聚合酶链式反应
Colon carcinoma
Proteins in regenerating liver cell-3
Invasion
RNA interference
Small interfering RNA
Fluorescence quantitative realtime polymerase chain reaction
