人肝内胆管癌细胞PRL-3的表达与侵袭转移研究  

Expression of Phosphatase of Regenerating Liver-3 in Intrahepatic Cholangiocarcinoma and its Role of Invasiveness and Metastasis

作　　者：徐亦君[1] 张振玉[1] 徐娴[1] 曹伟军[1] 竺明晨[2] 

机构地区：[1]南京医科大学附属南京医院消化科,江苏南京210006 [2]江苏省肿瘤医院检验科,江苏南京210009

出　　处：《现代生物医学进展》2012年第24期4642-4646,共5页Progress in Modern Biomedicine

基　　金：江苏省肿瘤医院青年创新研究项目(ZQ200904)

摘　　要：目的:探讨PRL-3在人肝内胆管癌侵袭转移中的作用。方法:利用小RNA技术干扰肝内胆管癌细胞株PRL-3表达,并采用细胞划痕实验和Transwell体外侵袭实验评价PRL-3对肝内胆管癌细胞侵袭转移能力的影响。结果:RT-PCR和Western blot结果均显示转染PRL-3特异性siRNA-2组PRL-3表达明显降低(P<0.05)。PRL-3 siRNA-2组在划痕培养24h后划痕区域宽度占初始划痕区域宽度的百分比为(62.12±6.28)%,阴性对照组为(23.88±2.55)%,空白对照HCCC-9810组为(21.20±6.07)%。PRL-3siRNA-2组细胞的划痕两端距离相比明显较宽,分别与阴性对照组细胞和空白对照HCCC-9810组细胞相比均有统计学意义(P<0.05),后两者无显著性差异(P>0.05)。Transwell体外侵袭实验结果显示,PRL-3 SiRNA-2组细胞侵袭能力明显减弱,穿膜细胞数为(19.40±2.30)个/HP,明显少于阴性对照组(64.00±2.73)个/HP和正常HCCC-9810组(67.20±3.11)个/HP,差异有显著性(P<0.05);正常HCCC-9810组和阴性对照组无明显差异(P>0.05)。结论:PRL-3特异性siRNA能够抑制肝内胆管癌细胞HC-CC-9810中内源性PRL-3的表达,并可以明显抑制肝内胆管癌细胞的迁移侵袭能力。Objective： To evaluate the role of phosphatase of regenerating liver-3 in intrahepatic cholangiocarcinoma invasiveness and metastasis. Methods： Human intrahepatic cholangiocarcinoma cell line, HCCC-9810 was transfected with PRL-3 siRNA. Scratch wound healing assay and transwell chambers were used to evaluate the motility and migrating velocity of cells. Results： The results of PCR and western blot showed that expression of PRL-3 significantly decreased （P〈0.05） after PRL-3 siRNA-2 transfection. Scratch wound healing results showed that the ratio of wound distance 24h after scratching and original wound distance in PRL-3 siRNA-2 cells, negative siRNA cells and parental HCCC-9810 cells, were respectively（ 62.12±6.28 ）%, （23.88± 2.55 ）% and （21.20± 6.07）%. The mi- grating distances of PRL-3 siRNA-2 transfecting cells was significantly shorter than those of negative siRNA cells and parental HC- CC-9810 cells （P〈0. 05）. In invasion assay, the number of cells that had migrated into the lower chamber were （ 19.40± 2.30）/HP in PRL-3 siRNA-2 group, （64.00 ±2.73 ）/HP in negative control group and （67.20 ±3.11 ）/lip in parental HCCC-9810 group. The differ- ence was significant between the group transfected with PRL-3 siRNA-2 and the other two groups （P〈0.05）. Conclusions： PRL-3 specific siRNA can inhibit the expression of PRL-3 in HCCC-9810 cells and weaken the invasion and migration potency of HCCC-9810 cells.

关 键 词：肝再生磷酸酶-3 RNA干扰 肝内胆管癌 转移 

分 类 号：R735.8[医药卫生—肿瘤]