沉默肝星状细胞神经菌毛素-1抑制肝癌生长的实验研究  

Silencing neuropilin-1 expression of hepatic stellate cells inhibits the growth of hepatocellular carcinoma

作　　者：徐之超[1] 陈晨[2] 慎浩鑫 马丽[2] 李文智[2] 王林[2] 耿智敏[2] 

机构地区：[1]西安交通大学第一附属医院急诊科,陕西西安710061 [2]西安交通大学第一附属医院肝胆外科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2016年第5期646-651,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30971340)~~

摘　　要：目的探讨沉默肝星状细胞神经菌毛素-1（neuropilin-1,NRP-1）表达后,是否会影响其对肝癌的促生长作用,并初步探讨可能存在的作用机制。方法细胞免疫荧光及细胞荧光双重染色观察NRP-1在肝星状细胞LX2中的表达及其与血小板源性生长因子受体β（platelet-derived growth factor receptor-β,PDGFR-β）的共表达;MTT法检测沉默肝星状细胞NRP-1后对肝癌细胞体外增殖能力影响;建立裸鼠皮下肝癌移植瘤模型,绘制生长曲线,免疫组化法观察各组瘤体α-平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）、NRP-1、PDGFR-β表达强度的差异。结果 LX2表面存在NRP-1表达,且NRP-1与PDGFR-β共表达于LX2;沉默LX2NRP-1后对HepG2促增殖作用降低（P〈0.05）;NRP-1KG移植瘤体积较NRP-1CG、NRP-1NG小（P〈0.05）,进一步对各组瘤体行免疫组化染色并进行表达强度积分分析,发现NRP-1KG瘤体间质NRP-1、PDGFR-β表达均较NRP-1CG弱（χ^2=25.89,P〈0.05;χ^2=28.12,P〈0.05）。结论沉默肝星状细胞NRP-1表达后,其对肝癌细胞促增殖作用降低,在体内环境中其对肝癌皮下移植瘤促生长作用也降低,其机制与肝星状细胞活化减弱有关,而肝星状细胞NRP-1表面共受体PDGFR-β表达减弱也可能参与这一过程。Objective To investigate whether the expression of hepatic stellate cells neuropilin-1 silenced（NRP-1）affects the growth of hepatocellular carcinoma cells（HCCs）and to explore the possible mechanism.Methods Expression of NRP-1 and co-expression with platelet-derived growth factor receptor-β（PDGFR-β）in hepatic stellate cells were observed by immunofluorescence and fluorescent double staining.The effect of silencing NRP-1 of HSCs on the proliferation of HCC cells was detected by MTT in vitro.A xenograft hepatocellular carcinoma model of nude mouse was established subcutaneously.The growth curve was drawn and the expressions ofα-SMA,NRP-1 and PDGFR-βin tumors were observed by immunohistochemistry staining after the mice were killed.Results NRP-1 was expressed in the membrane of LX2 and was co-expressed with PDGFR-β.Silencing NRP-1 of LX2 reduced the proliferation of HepG2 in vitro（P〈0.05）.The tumor volume in NRP-1 KG group reduced obviously compared with that in NRP-1 CG group（P〈0.05）.The expressions of NRP-1 and PDGFR-βin NRP-1 KG group were weaker than those in NRP-1 CG group by immunohistochemistry（χ^2=25.89,P〈0.05;χ^2=28.12,P〈0.05）.Conclusion Silencing NRP-1 expression of HSCs can decrease its effect on the proliferation of HCCs in vitro and the growth-promoting effect of HSCs on HCCs is also decreased in vivo,which is due to the inhibition of HSCs activation,and the decreased expression of co-receptor PDGFR-βmay play a role in this process.

关 键 词：神经菌毛素-1 肝星状细胞 肝癌 肿瘤微环境 

分 类 号：R735.7[医药卫生—肿瘤]