ShRNA靶向沉默FGFR3基因对人肝癌细胞Huh7增殖和迁移的影响  

Effect of FGFR3 gene silenced by shRNA on proliferation and migration of human liver cancer Huh7 cell

作　　者：李科[1] 杨卫平[1] 付志平[1] 汪炳瑞 景晓乾 沈柏用[1] 彭承宏[1] 邱伟华[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院外科上海消化外科研究所,上海200025

出　　处：《外科理论与实践》2014年第4期342-348,共7页Journal of Surgery Concepts & Practice

基　　金：国家自然科学基金(81172326);上海慈善癌症研究基金

摘　　要：目的:研究成纤维细胞生长因子受体3(FGFR3)对肝癌细胞株Huh7增殖和迁移能力的影响。方法:以靶向沉默FGFR3的shRNA慢病毒表达载体、delta8.9和VSVG三质粒共转染293T细胞来包装慢病毒颗粒。选择Huh7细胞转导慢病毒载体。通过细胞增殖实验(CCK-8法)和transwell实验,分别评估沉默FGFR3对Huh7肝癌细胞增殖和迁移能力的影响。裸鼠分别皮下注射pSilencer-FGFR3-shRNA1#组和pSilencer-NC组Huh7细胞后,监测肿瘤生长。Western印迹法检测下游信号蛋白p-AKT和p-ERK。结果:与NC组比较,RNAi组的细胞增殖能力显著下降(P<0.01),裸鼠皮下移植瘤体积较小[shRNA1#组:(210.2±94.6)mm3,NC组:(1 546.0±331.7)mm3,P<0.05];迁移能力亦显著下降[shRNA1#组:(6.3±1.2)个,shRNA2#组:(3.0±0.5)个,NC组:(36.7±4.4)个,P<0.001]。RNAi组p-ERK和p-AKT的蛋白质表达水平显著下降。结论:沉默FGFR3,可能通过抑制ERK和AKT通路,显著降低肝癌细胞的增殖和迁移能力。Objective To investigate the effect of FGFR3 on the proliferation and migration in liver cancer cell lines Huh7. Methods ShRNA lentiviral expression vectors targeting human FGFR3 were constructed and co-transfected into 293T with delta8.9 and VSVG to package lentivirus particles. Huh7 cells were transduced with the constructed lentiviral vectors. The effect of FGFR3 on cell proliferation and migration was investigated by CCK-8 assay and transwell assay. Huh7 cells transfected with pSilencer-FGFR3-shRNA1# and pSilencer-NC （negtive control） were subcutaneously injected into nude mice respectively. Tumor growth was monitored. The expression of p-AKT and p-ERK in Huh7 cells were detected by Western blot. Results Compared with negative control group （NC）, the proliferation in RNAi groups was significantly suppressed （P〈0.01）, the volume of xenograft in nude mice was smaller [shRNA1#： （210.2±94.6） mm^3, NC：（1 546.0±331.7） mm3, P〈0.05]; the ability of migration was also significantly suppressed [shRNAI#：（6.3±1.2）, shRNA2#：（3.0±0.5）, NC： （36.7±4.4）, P〈0.001]. The amount of protein of p-ERK and p-AKT significantly decreased in RNAi groups. Conclusions Silencing of FGFR3 could significantly inhibit cell proliferation and migration via suppressing ERK and AKT signaling pathway.

关 键 词：肝细胞癌 FGFR3基因 SHRNA 增殖 迁移 

分 类 号：R735.7[医药卫生—肿瘤]