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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。
AIM: To investigate the effect of fibroblast growth factor receptor 3 (FGFR3) on the proliferation and migration of hepatocellular carcinoma cell line Huh7. METHODS: Lentiviral shRNA expression vectors targeting shRNA targeting silencing FGFR3 were constructed and 293T cells were co-transfected with the delta8.9 and VSVG plasmids. Huh7 cells are selected for transduction of lentiviral vectors. The effects of silencing FGFR3 on the proliferation and migration of Huh7 hepatoma cells were evaluated by cell proliferation assay (CCK-8 assay) and transwell assay. Tumor growth was monitored after subcutaneous injection of pSilencer-FGFR3-shRNA1 # and pSilencer-NC Huh7 cells in nude mice, respectively. Western blotting was used to detect the downstream signaling proteins p-AKT and p-ERK. Results: Compared with NC group, the cell proliferation ability of RNAi group was significantly decreased (P <0.01), and the volume of subcutaneous tumor in nude mice was smaller than that of NC group [shRNA1 #: (210.2 ± 94.6) mm3, NC group: (5446.0 ± 331.7) mm3, P <0.05]. The migration ability was significantly decreased as well. [shRNA1 # group: (6.3 ± 1.2), shRNA2 # group: (3.0 ± 0.5), NC group: (36.7 ± 4.4), P <0.001; The protein expression levels of p-ERK and p-AKT in RNAi group were significantly decreased. Conclusion: Silencing FGFR3 may significantly reduce the proliferation and migration of hepatocellular carcinoma cells by inhibiting ERK and AKT pathways.