摘要
目的:探讨纤维胶凝蛋白3(FCN3)在肝细胞癌(HCC)组织中的表达及对HCC细胞增殖、迁移和上皮-间质转化(EMT)的影响。方法:首先利用生物信息学技术分析FCN3在HCC中的表达及与患者预后的关系;然后通过qRT-PCR、免疫组化进一步检测FCN3在HCC细胞和组织中的表达。构建过表达FCN3的Huh7稳转细胞株,通过CCK-8实验、细胞划痕实验和Transwell实验检测FCN3对HCC细胞增殖、迁移、侵袭的影响;通过Western Blot实验检测E-cadherin、N-cadherin、Vimentin表达水平;通过裸鼠皮下成瘤实验,在体内探究FCN3对HCC增殖的影响。结果:生物信息学分析表明FCN3在HCC组织中的表达水平显著低于癌旁组织,高表达FCN3的HCC患者具有更长的总体生存时间。qRT-PCR表明,与正常肝细胞相比,HCC细胞株Huh-7、Hep3B和SNU-449中FCN3 mRNA表达水平显著降低。免疫组化表明,FCN3在HCC组织中染色阳性细胞的比例显著低于癌旁组织。选取FCN3相对表达量最低的Huh-7细胞株进行慢病毒稳转FCN3基因,得到稳定OE-Huh7和空载慢病毒稳转的NC-Huh7。Transwell实验、划痕实验、CCK8实验表明,与NC-Huh7组相比,OE-Huh7组的迁移、增殖、侵袭能力显著降低;Western Blot结果显示,OE-Huh7组Vimentin、N-cadherin蛋白表达量降低,E-cadherin蛋白表达量升高。结论:FCN3在HCC中低表达且影响患者的预后,过表达其水平后可抑制HCC细胞的增殖、迁移及EMT。
Objective:To investigate the expression of Ficolin-3(FCN3)in hepatocellular carcinoma(HCC)and its effects on cell proliferation,migration and epithelial-mesenchymal transformation(EMT).Methods:Firstly,bioinformatics was used to ana-lyze the expression of FCN3 in HCC and its relationship with the prognosis of patients.Then,the expressions of FCN3 in HCC cells and tissues were further detected by qRT-PCR and immunohistochemistry.Huh7 stable cell lines overexpressing FCN3 were constructed,and the effects of FCN3 on the proliferation,migration and invasion of HCC cells were detected by CCK-8,scratch and Transwell assays.The expression levels of E-cadherin,N-cadherin and Vimentin were detected by Western Blot assay.The effect of FCN3 on HCC proliferation was investigated in vivo by subcutaneous tumor formation in nude mice.Results:Bioinfor-matics analysis showed that the expression level of FCN3 in HCC tissues was significantly lower than that in paracancer tissues,and HCC patients with high expression of FCN3 had longer overall survival time.qRT-PCR showed that mRNA expression levels of FCN3 in HCC cell lines Huh-7,Hep3B and SNU-449 were significantly decreased compared with normal hepatocyte.Immuno-histochemistry showed that the proportion of FCN3 staining positive cells in HCC tissues was significantly lower than that in para-cancer tissues.The Huh7 cell line with the relatively lowest expression of FCN3 was selected for lentivirus stable FCN3 gene,and stable OE-Huh7 and no-load lentivirus stable NC-Huh7 were obtained.Transwell,scratch and CCK8 tests showed that the migra-tion,proliferation and invasion ability were significantly reduced in OE-Huh7 group compared with NC-Huh7 group.Western Blot results showed that the expression levels of Vimentin and N-cadherin protein decreased while the expression levels of E-cadherin protein increased in OE-Huh7 group compared with NC-Huh7 group.Conclusions:The expression of FCN3 is low in HCC,and FCN3 affects the prognosis of patients.Overexpression of FCN3 can inhibit the pro
作者
龚旋坤
包凌
周帅
庞青
GONG Xuankun;BAO Ling;ZHOU Shuai;PANG Qing(Department of Hepatobiliary Surgery,The First Affliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Geriatrics,The First Affliated Hospital of Bengbu Medical College,Bengbu 233000,China;The Fifth Clincal Medical College,Anhui No.2 Provincal People’s Hospital,Anhui Mediacl University,Hefei 230041,China)
出处
《海南医学院学报》
CAS
北大核心
2024年第7期498-504,共7页
Journal of Hainan Medical University
基金
国家自然科学基金青年项目(81600452)
安徽省自然科学基金杰青项目(2008085J37)。
关键词
肝细胞癌
FCN3
上皮-间质转化
增殖
迁移
Hepatocellular carcinoma
FCN3
Epithelial-mesenchymal transformation
Proliferation
Migration
