沉默成纤维生长因子受体4基因对宫颈癌Hela细胞生物学特性的影响  

Study on the effect of FGFR4 gene silencing on biological properties of Hela cervical cancer cells

作　　者：李亚萍[1] 邹余粮[2] 

机构地区：[1]西安市中心医院妇产科,陕西西安710003 [2]西安交通大学第一附属医院妇产科,陕西西安710061

出　　处：《海南医学院学报》2017年第20期2749-2752,2756,共5页Journal of Hainan Medical University

基　　金：陕西省科学技术研究发展计划项目(2010K01-161)~~

摘　　要：目的:探讨沉默成纤维细胞生长因子受体4(FGFR4)对宫颈癌Hela细胞生长、转移以及顺铂(DDP)化疗敏感性产生的影响。方法:培养宫颈癌Hela细胞系并分为5组:空白对照组、阴性对照组、FGFR4-siRNA组、DDP组和FGFR4-siRNA+DDP组,设计合成靶向FGFR4的siRNA转染FGFR4-siRNA组和FGFR4-siRNA+DDP组细胞,而后对DDP组和FGFR4-siRNA+DDP组加入化疗药物DDP(10ug/mL)。分别于培养24h、48h和72h后MTT法检测各组细胞生长抑制率,流式细胞仪检测凋亡率;PCR检测FGFR4以及肿瘤细胞生长、转移相关基因VEGF和MMP2的mRNA水平,Western blot检测其蛋白表达水平。结果:沉默FGFR4基因后,与对照组相比,FGFR4-siRNA组细胞增殖活性下降,生长抑制率和细胞凋亡率上升,FGFR4几乎不表达,VEGF和MMP2表达水平下降;FGFR4-siRNA+DDP组细胞生长抑制率和凋亡率高于FGFR4-siRNA组和DDP组,FGFR4几乎不表达,VEGF和MMP2表达水平均低于FGFR4-siRNA组和DDP组。结论:沉默FGFR4基因的表达能够抑制宫颈癌细胞生长,促进细胞凋亡和提高对顺铂的化疗敏感性,下调VEGF和MMP2的表达抑制癌细胞的侵袭转移能力,可以作为宫颈癌新的靶向治疗目标之一。Objective：To discuss the the effect of FGFR4 gene silencing on growth,metastasis and the cis-platinum（DDP）chemotherapy sensitivity of Hela cervical cancer cells.Methods：The Hela cervical cancer cells were cultured and divided into 5 groups：blank control group,negative control group,FGFR4-siRNA group,DDP group and FGFR4-siRNA＋DDP group.The siRNA targeted to FGFR4 was constructed and transfected into the cells in FGFR4-siRNA group and FGFR4-siRNA＋DDP group,and DDP（10 ug/mL）was added into the DDP group and FGFR4-siRNA＋DDP group.The growth inhibition ratio was detected by MTT and the apoptosis rate was detected by FCM after 24 h,48 hand 72 hculturing respectively.The mRNA and protein expression level of FGFR4,VEGF and MMP2 were detected by RT-PCR and western blot respectively.Results：Compared with the blank control,the proliferative activity of FGFR4-siRNA group decreased after the FGFR4 silenced,and the growth inhibition ratio and apoptosis rate increased,the FGFR4 did almost not express and the expression level of VEGF and MMP2 decreased.The growth inhibition ratio and apoptosis rate of FGFR4-siRNA＋DDP group were higher than that of FGFR4-siRNA group and DDP group,the FGFR4 did almost not express either and the the expression level of VEGF and MMP2 were lower than the FGFR4-siRNA group and DDP group.Conclusion：It can be obtained that silencing the FGFR4 of cervical cancer could inhibit cell growth,promote cell apoptosis and increase the chemosensitivity,down-regulate the expression of VEGF and MMP2 to inhibit the ability of invasion and metastasis,which could be one of the targeted therapeutic targets of cervical cancer.

关 键 词：宫颈癌 成纤维生长因子受体4 SIRNA HELA细胞 顺铂 

分 类 号：R737.33[医药卫生—肿瘤]